Fernando Moraes
email: moraes@inf.pucrs.br

Michel Robert
email: robert@lirmm.fr

Daniel Auvergne
email: auvergne@lirmm.fr

Nadine Azemard
email: azemard@lirmm.fr

A Physical Synthesis Design Flow Based on Virtual Components

This paper presents a virtual library design flow for automatic layout synthesis tools. The motivation to develop such design flow is to enable the use of static CMOS complex gates (SCCGs), to optimize area and delay at the logic and physical abstraction levels. The use of SCCGs increases the design space (number of different primitive cells) when compared to the state-of-the-art cell based approaches. Therefore, a new design flow is defined, which replaces cell libraries by virtual components, creating in this way a virtual library. We present the procedure to create such virtual library in the Synopsys environment, and to integrate logic synthesis tools in automatic layout synthesis.

INTRODUCTION

Previous publications ( [START_REF] Lefebvre | The Future of Custom Cell Generation in Physical Synthesis[END_REF], [START_REF] Ienne | Practical Experiences with Standard-Cell Based Datapath Design Tools -Do We Really Need Regular Layouts? DAC'98[END_REF], [START_REF] Burns | C5M -A Control-Logic Layout Synthesis System for High-Performance Microprocessors[END_REF]) have presented the advantages/disadvantages of using traditional cell based design or tools for automatic layout synthesis.

Briefly, cell based design has a well established design flow (from behavioral VHDL), cells are precharacterized and area/delay figures can be computed at the gate level. However, it is not possible to size the transistor widths (the solution in libraries is to provide several templates for each cells, increasing the library complexity/ management) and the library is designed for one given fabrication process. Considering deepsubmicron processes, where wire delay must be considered, it should be possible to size each gate according to the load to be driven.

Following [START_REF] Lefebvre | The Future of Custom Cell Generation in Physical Synthesis[END_REF][START_REF] Ienne | Practical Experiences with Standard-Cell Based Datapath Design Tools -Do We Really Need Regular Layouts? DAC'98[END_REF], cell synthesis from transistor netlist level is a flexible way to support open library architecture and satisfy timing closure on cell based design.

In [START_REF] Lefebvre | The Future of Custom Cell Generation in Physical Synthesis[END_REF] we have: "Cell synthesis is concerned with creating cell layouts starting with only a transistor level netlist for each cell. It is completely flexible in terms of a target library architecture and does not require any pre-existing cell-specific layout information." In [START_REF] Ienne | Practical Experiences with Standard-Cell Based Datapath Design Tools -Do We Really Need Regular Layouts? DAC'98[END_REF] we have: "...it clearly appears that on timing-critical cell-based designs it is nowadays a severe penalty not to use a synthesizer."

Then, one alternative to the cell-based approaches is to automatically generate the layout of each gate, without using cell libraries. Using such approach, each gate can be individually sized, and synthesis will be independent of process fabrication.

Automatic layout synthesis tools to be effectively used by industry or IP providers must solve the following problems: (i) how to define a virtual library; (ii) how integrate automatic layout synthesis with a logic synthesis tool, using a non limited virtual library; (iii) how to characterize the virtual library to allow delay prediction at the gate level. This paper presents a solution for the first two questions: definition of the virtual library and the integration of layout and logic synthesis tools. Solution for library characterization has been presented in [START_REF] Daga | A Comprehensive Delay Macro-Model of Submicrometer CMOS Logics[END_REF] and will be used as necessary.

Section 2 presents the conceptual design flow using a virtual library methodology. Section 3 presents the implementation of such methodology using Synopsy [4] as the logic synthesis environment. Section 4 presents the physical synthesis [START_REF] Moraes | A Virtual CMOS Library Approach for Fast Layout Synthesis[END_REF], as well as the optimization step of the design flow.

LIBRARY FREE TECHNOLOGY MAPPING

The concept of a virtual library is based on using cells available through a layout generator, instead of using a set of pre-characterized cells.

Due to design and maintenance cost, the number of cells available into a library is quite small when compared to the possibilities given by using complex gates (or SCCGs -Static CMOS Complex Gates). The number of different functions that can be implement with SCCGs having 4 or less transistors connected in series is 3503 ( [START_REF] Detjens | Technology mapping in MIS[END_REF]) and this number grows to 425803 for arrays of 5 transistors. So, the use of SCCGs available through layout generation increases the design space (number of different primitive cells) when compared to state-ofthe-art cell based approaches.

The set of available SCCGs is defined by user constrains, e.g., number of transistors in series, maximum fanout and output isolation. These topological restrictions define the virtual library to be used in library free design approaches. Therefore, virtual library approach means the possibility to use any kind of SCCG for a given circuit, with individual transistor sizing.

Design Flow

Figure 1 shows the design flow for the virtual library approach. The circuit is specified using a behavioral/structural VHDL

User constraints

Figure 1 -Design flow in the virtual library approach

This structural description is automatically converted to a SPICE format, since for the automatic layout synthesis a transistor level description is required. The transistors are initially sized with a constant width. The width is a multiple of the minimum transistor width for the employed technology.

A first layout synthesis is performed (Tropic in the figure), aiming parasitic capacitance evaluation. The transistor level description and the parasitic capacitances are used for timing analysis and electrical optimization. If timing constraints are not met, some optimization strategies can be employed: transistor sizing and/or buffer/repeater insertion.

After electrical optimization, if needed, a second layout synthesis is made, resulting in the final layout of the circuit, with each transistors individually sized. It is important to keep the same placement between both layout syntheses, in order to get the same routing and consequently the same parasitic capacitances.

VIRTUAL LIBRARY APPROACH IN THE SYNOPSYS ENVIRONMENT

We present in this section a practical implementation of a virtual library approach.

Figure 2 illustrates the first part of the flow, which consists to generate a structural VHDL form a behavioral description, using a virtual library during the technology mapping.

Virtual.LIB BEHAVIORAL VHDL

LIBRARY COMPILER (SYNOPSYS)

Virtual.DB Synthesis script

DESIGN COMPILER (SYNOPSYS)

STRUCTURAL VHDL SPICE FILE

Figure 2 -Virtual library approach in the Synopsys environment

The first action is to describe this virtual library. For example, Figure 3 shows a SCCG description in the Synopsys format. Each SCCG have: (1) its name, which is used in the final structural VHDL; (2) the area, which is proportional to the number of inputs (this is true, since a linear matrix layout style is used by layout synthesis tools [START_REF] Moraes | A Virtual CMOS Library Approach for Fast Layout Synthesis[END_REF] In this example a very simple time model, the generic CMOS [8], is used. Work under development consists in defining a complete table lookup model. Each SCCG will be characterized from a set of delay equations for sub-micron technology [START_REF] Daga | A Comprehensive Delay Macro-Model of Submicrometer CMOS Logics[END_REF]. In this way, the delay prediction at the logic level will have the same accuracy of the cell-based approaches.

A virtual library is automatically generated from the technology data (electrical model) and topological constraints. No electrical simulations or layout generations are required to perform this task. There is no restriction on the number of cells into the virtual library, all CMOS static SCCG can be described.

The sequential elements, flip-flops, latches and tristates are also inserted in the virtual library file. These sequential elements are not generated automatically, due to its complex description. A template file is modified, according to the technology data. The sequential library has 9 components: 4 FFDs (only D, set, reset, set/rest), 4 LATCHs (only D, set, set/rest) and 1 buffer tri-state.

Once the virtual library file generated, it is compiled using the "library compiler" [8] tool (Synopsys), resulting in a library file accepted by the logic synthesis tool, "design compiler" (Synopsys).

The input VHDL description can be behavioral, structural or a mixed of them. If a behavioral VHDL (Figure 4-a) is used as input, the designer describes his circuit without references to the cells inside the virtual library. or mixed description is used, the designer must known in advance the cell names, since the logic synthesis will make the mapping using the cell names of the description. In this example, the cells nor3, inv and AOI_BK3 must be in the virtual library, since they are instanced (command port map).

A synthesis script, containing the area and timing constraints, guides the logic synthesis. Figure 5 shows a synthesis script, where area (set_max_area), fanout (set_max_fanout) and delay constraints (set_max_delay) are imposed. It is important to generate a flat output description (commands uniquify and ungroup). This is a constraint imposed by the layout synthesis tool. write -format vhdl -hierarchy -output cell + "_S.vhdl" } quit

Figure 5 -Synthesis script, with delay and area constraints

The structural VHDL, synthesized by Synopsys and mapped over our virtual library, is automatically converted to a SPICE format. The final SPICE netlist has three components: sub-circuits, reference to the sub-circuits and output/input pins.

Each cell (component, in VHDL) used in the structural VHDL is translated into a spice subcircuit, using the command function (see Figure 3). Figure 6 shows a cell equation, and the respective spice sub-circuit, automatically generated. The translation of an equation to a sub-circuit is performed by a recursive function, which considers the operation '*' (and) as transistors connected in series and the operation '+' as transistors connected in parallel, for the N plan (the P plan is dual to the N plan). Observe that all functions in the virtual library are negative. The tool responsible to translate the netlist spice from structural VHDL reads the same file used by the library compiler tool. The sequential elements are also created from templates (nine previously defined spice subcircuits are used).

All references to cells (port map, in VHDL) used in the structural VHDL is translated sub-circuit calls. Finally, the input/output pins are obtained from the entity declaration. The resulting SPICE file is used as input description for our layout synthesis tool, TROPIC3.

Example of an initial VHDL and final SPICE files are shown in Figure 7.

!((A*B*C)+(D*E*F)+(G*H*I))

SUBCKT

PHYSICAL SYNTHESIS

This Section presents briefly the layout synthesis features and the work under development in timing analysis/optimization.

The layout synthesis tool, TROPIC [START_REF] Moraes | A Virtual CMOS Library Approach for Fast Layout Synthesis[END_REF], is a macrocell generator. The initial SPICE description, obtained from a behavioral VHDL, is decomposed into leaf cells that will be assembled together by dedicated place and route tools, without constitute a separate library. Two instances of the same logic function can have different layouts, according to its environment.

The main features of the layout style:

• Linear-matrix layout style, each cell row is composed of two horizontal diffusion strips; • Routing is implemented with 3 metal layers and stacked contacts, reducing the routing area; • No layout compaction. This is the main feature of TROPIC, since it allows a very fast layout synthesis. Tools, like LAS [START_REF] Cadence | Virtuoso layout synthesizer -LAS -user guide[END_REF], create an intermediate symbolic layout description, requiring layout compaction, consuming a lot of CPU time. • Complete parasitic capacitance/resistance evaluation.

• Simple technology file to describe design rules (28 rules) and parasitic capacitances/resitances (26 rules).

At the cell level, we can enumerate the following features (Figure 8):

• connection between N and P plan directly in metal 1; • minimum separation between N and P;

• gnd/vcc wires between transistors, in metal2;

• over-the-cell routing (OTC), connecting internal nodes of SCCGs and nets belonging to only one channel; • jogs are automatically inserted in the polysilicon wires, to reduce diffusion area (capacitance reduction); • polysilicon gate is only aligned to a virtual grid if it is connect to the routing region of the circuit. The layout generator has only 2 input files: the SPICE netlist and the design rules file. Its outputs are the layout (CIF format) and the parasitic capacitance/resistance estimation (flat spice netlist).

Figure 9 shows some transistor densities for TROPIC for 2 different processes. For a 0,25 µm process, an average transistor density of 60000 transistors/mm 2 was obtained. Refinements in the routing algorithm can improve this density up to 20% [START_REF] Moraes | A Virtual CMOS Library Approach for Fast Layout Synthesis[END_REF]. The CPU time, to generate the layout and estimate parasitic capacitances, for the largest example (15000 transistors), was 227,67 seconds (in an Ultra-10 Sparc). As CPU time for layout generation is no more a bottleneck, iterations can be made to obtain an optimized circuit. The logic synthesis tool can execute initial iterations to get accurate information on routing length (parasitic capacitances) and area. This can guide the technology mapping and also indicate where buffers must be inserted, since the real load in each node is calculated during the layout synthesis.

There are three capacitance components: line-toground, line-to-line and crossover capacitances. The line-to-ground and crossover capacitances are calculated by the traditional formulation based in the area and perimeter of the interconnections. We used a simple and accurate 2½D methodology described in [START_REF] Cong | Analysis and Justification of a Simple, Practical 2 1/2D Capacitance Extraction Methodology[END_REF]. For each connection in the layer i, we analyze the immediate neighbor in the same layer, all crossunders in the layer i-1 and all crossovers in the layer i+1, treating the layers i±2 like ground planes. However, the line-to-line capacitance is a function of the distance between two connections and the thickness of the layer. We use the empiric formulation described in [START_REF] Chern | Multilevel Metal Capacitances Models for CAD Design Synthesis Systems[END_REF].

The components of the diffusion capacitance, the area and perimeter of drain/source areas also computed by the layout generator.

The circuit delay obtained through electrical simulation using the evaluated capacitances was compared to extracted capacitances. Preliminary results show an average difference of only 5% between estimation and extraction.

The resistances are calculated using the number of squares in each wire, plus the number of contacts and vias. This simple model is been improved. If resistances are also evaluated, the user can choose between 3 representation models (with one state): L, T and Pi.

Electrical Optimization

After layout generation and capacitance evaluation, the last step in the design flow is the timing analysis and electrical optimization.

At the physical step of the design, the delay and power performances of MOS integrated circuits can be controlled by the appropriate selection of transistor sizes. This sizing problem is generally addressed as a non-linear optimization problem with a number of parameters equal to the number of transistors to be considered. Alternatives can be obtained by regularly varying the transistor drive capabilities until the longest and shortest paths fulfill the delay-power constraint [START_REF] Cremoux | Path resizing based on incremental technique[END_REF]. The consideration on the sensitive paths of the drive capability of gates may reduce the global optimization problem to local optimizations allowing effective management of the circuit delay and power.

The satisfaction of imposed delay constraints implies a full path enumeration with a complete account of the real (post layout) evaluation of the performance parameters of the different switching blocks. For that, it is necessary to get available fast and accurate path identification techniques. This has been obtained using graph exploring techniques such as BFS or DFS algorithms. However in these enumeration techniques all the circuit paths are considered and stored at the expense of prohibitive CPU time and memory allocation when considering large circuits. As a result no complete path enumeration and classification, allowing speedpower trade-off on the different branches can be performed.

The problem of circuit performance optimization must be addressed on a complete path enumeration.

We considered an incremental technique, which supplies the enumeration of the longest (shortest) paths in a decreasing (increasing) delay order.

One advantage of this technique is to work on a user specified limited number of paths [START_REF] Yen | Efficient Algorithms for Extracting the k Most Critical Paths in Timing Analysis[END_REF], allowing the easy application of different path optimization criteria. For that we developed a sizing algorithm to satisfy delay (power) constraints. This algorithm size the gates belonging to the paths classified by the incremental technique. The selected criterion for gate resizing is defined through the gate load to drive ratio, evaluated on each node and which has been shown to constitute a robust metric for the gate strength and the gate loading evaluation [START_REF] Yen | Efficient Algorithms for Extracting the k Most Critical Paths in Timing Analysis[END_REF]. This parameter associated to the physical performance description of the gate load (considering the estimated parasitic capacitances), gives direct indication of its loading level. Results of such work can be found in [START_REF] Cremoux | Path resizing based on incremental technique[END_REF].

DISCUSSION

We have presented in this paper a complete design flow for automatic layout synthesis tools, coupled with logic synthesis and electrical optimization. Our main contributions are (i) the definition of the virtual library using SCCGs and (ii) the integration of the logic synthesis to the physical synthesis. The characterization of the virtual library is a work under development.

Another alternative to the virtual library was investigated. A simple library (containing basic gates: and, or, inverters) is used during the logic synthesis, resulting in an intermediate description. This description is used by a specific tool for technology mapping, like SIS, generating the SPICE netlist with complex gates. This approach was rejected, since there is no way to control design constraints at the logic level because only simple gates are used.

The advantages of virtual libraries over cell libraries are:

• great flexibility during the technology mapping;

• realistic parasitic evaluation without electrical extraction; • an incremental path enumeration allowing transistor sizing according to the user constraints; • easy technology migration. Design flow based on virtual libraries can replace cell-based approaches in deep sub-micron technologies. This approach allows a fast technology independent IP prototyping for the design of systems on silicon.

Figure 3 -

 3 Figure 3 -SCCG described in the Synopsys format (simplified timing)

( a )

 a Figure 4 -Behavioral and structural VHDL descriptions

  link_library = {sccg33.db libdff.db} target_library = {sccg33.db libdff.db} cell_list = { sklansky } foreach (cell, cell_list) { read -f vhdl cell + ".vhdl"

(b) Spice file automatically obtained from the input VHDLFigure 7 -

 7 Figure 7 -Original VHDL description and resulting SPICE file

Figure 8 -

 8 Figure 8 -Layout example (row level)

Figure 9 -

 9 Figure 9 -Transistor densities for TROPIC

SPICE Behavioral VHDL Struct. VHDL VIRTUAL LIB Logic functions Technology data TROPIC SYNOPSYS Conversion Struc. VHDL è è è è SPICE LAYOUT OPTIMIZATION: Sizing, buffers, repeaters TIMING ANALYSIS TROPIC Parasitic capacitances

	description. A logic
	synthesis tool synthesizes the circuit (Synopsys in
	the figure), generating as result a structural VHDL.
	A virtual library containing a rich set of SCCGs is
	used during the technology mapping. User
	constraints, like critical path delay, output loads,
	fanout restrictions are specified in the synthesis
	script.

SCCG equation and its spice description

					AOI1 out A B C D E F G H I vcc
				MN1	gnd	I	6 NMOS w=2.0um l=0.25um
				MN2	6	H	3 NMOS w=2.0um l=0.25um
				MN3	3	G out NMOS w=2.0um l=0.25um
				MN4	gnd	F	5 NMOS w=2.0um l=0.25um
				MN5	5	E	2 NMOS w=2.0um l=0.25um
				MN6	2	D out NMOS w=2.0um l=0.25um
				MN7	gnd	C	4 NMOS w=2.0um l=0.25um
				MN8	4	B	1 NMOS w=2.0um l=0.25um
				MN9	1	A out NMOS w=2.0um l=0.25um
				MP10	vcc	I 16 PMOS w=2.0um l=0.25um
				MP11	vcc	H 16 PMOS w=2.0um l=0.25um
				MP12	vcc	G 16 PMOS w=2.0um l=0.25um
				MP13	16	F 15 PMOS w=2.0um l=0.25um
				MP14	16	E 15 PMOS w=2.0um l=0.25um
				MP15	16	D 15 PMOS w=2.0um l=0.25um
				MP16	15	C out PMOS w=2.0um l=0.25um
				MP17	15	B out PMOS w=2.0um l=0.25um
				MP18	15	A out PMOS w=2.0um l=0.25um
				.ends AOI1
	Figure 6 -Entity DIV2 is ** SUBCIRCUITS (generated from the lib file)
	Port(clk,rst: in std_logic; d2_clk: out std_logic );	.SUBCKT DFFR D rst clk QN Q vcc
	end DIV2;			MN1	H clk gnd gnd NMOS l=0.25u w=2u
	Architecture arch of DIV2 is		MP2	H clk vcc vcc PMOS l=0.25u w=2u
	signal temp : std_logic;		MN3	NH	H gnd gnd NMOS l=0.25u w=2u
	begin			MP4	NH	H vcc vcc PMOS l=0.25u w=2u
	d2_clk <= temp;			…
	process (clk,rst)			.ends DFFR
	begin
	if rst='0' then	temp<='0';		.SUBCKT GAT1_1	out A vcc
	elsif clk'event and clk='1' then temp<=not temp;	MN1	gnd	A out gnd NMOS l=0.25u w=2u
	end if;			MP2	vcc	A out vcc PMOS l=0.25u w=2u
	end process;			.ends GAT1_1
	end;
				**	COMPONENTS (generated from the VHDL file)
	Entity div4 is			X1 net21 n3 clk net21 div vcc DFFR
	Port(clk,rst:in std_logic; div,div4,div8: out std_logic );	X2 net20 n3 n1 net20 div8 vcc DFFR
	end div4;			X3 net19 n3 n2 net19 div4 vcc DFFR
	Architecture arch of div4 is		X4 n3 rst vcc GAT1_1
	signal carry : std_logic_vector(2 downto 0);	X5 n2 net21 vcc GAT1_1
	begin			X6 n1 net19 vcc GAT1_1
	st0: DIV2 Port Map(clk=>clk,	rst=>rst, d2_clk=>carry(0));
	st1: DIV2 Port Map(clk=>carry(0),rst=>rst, d2_clk=>carry(1));	** INPUTS AND OUTPUTS (generated from ENTITY in the VHDL file)
	st2: DIV2 Port Map(clk=>carry(1),rst=>rst, d2_clk=>carry(2));	*interface: div8	orient n output
	div <= carry(0);			*interface: div4	orient n output
	div4 <= carry(1);			*interface: div	orient n output
	div8 <= carry(2);			*interface: rst	orient s input
	end;			*interface: clk	orient s input
	(a) Input description: VHDL

Acknowledgements

Fernando Moraes gratefully acknowledges the support of the CNPq through grant no. 522939/96-1.


Giorgia Battipaglia
email: giorgia.battipaglia@aphp.fr

Florent Malard

Marie Thérèse Rubio

Annalisa Ruggeri

Anne Claire Mamez

Eolia Brissot

Federica Giannotti

Remy Dulery

Anne Christine Joly

Minh Tam Baylatry

Marie Therése Rubio

Marie Jeanne Kossman

Jaques Tankovic

Laurent Beaugerie

Harry Sokol

Mohamad Mohty

Marie Therése Rubio

Anne Claire Mamez

Marie Therèse Rubio

Marie Jeanne Kossmann

Jacques Tankovic

Fecal microbiota transplantation before or after allogeneic hematopoietic transplantation in patients with hematologic

à la diffusion de documents scientifiques de niveau recherche, publiés ou non, émanant des établissements d'enseignement et de recherche français ou étrangers, des laboratoires publics ou privés.

Introduction

During the last decades, the prevalence of multidrug resistant bacteria (MDRB), has largely increased, becoming a serious worldwide problem [START_REF] Spellberg | The Epidemic of Antibiotic-Resistant Infections: A Call to Action for the Medical Community from the Infectious Diseases Society of America[END_REF] . Under physiological conditions, commensal microbiota prevents gut colonization from MDRB. However, in particular conditions, such as in patients with hematologic malignancies, use of chemotherapeutic agents and broad spectrum antibiotics may favor selection of resistant pathogens through the alterations of the gastrointestinal barrier and the consequent dysbiosis [START_REF] Montassier | Chemotherapy-driven dysbiosis in the intestinal microbiome[END_REF] . Patients undergoing allogeneic hematopoietic stem cell transplantation (allo-HSCT) are at even higher risk of dysbiosis due to their profound immune depression [START_REF] Samet | Leukemia and risk of recurrent Escherichia coli bacteremia: Genotyping implicates E. Coli translocation from the colon to the bloodstream[END_REF] . In case of bloodstream infections from MDRB, outcomes are even poorer, with consequently increased mortality [START_REF] Bilinski | Impact of Gut Colonization by Antibiotic-Resistant Bacteria on the Outcomes of Allogeneic Hematopoietic Stem Cell Transplantation: A Retrospective, Single-Center Study[END_REF] . An Italian study, for example, showed that carbapenemase producing (CP-) bacteria, including Pseudomonas aeruginosa, were independent predictors of death in patients diagnosed with acute leukemia, while this was not observed in case of extended-spectrum β-lactamase (ESBL-) Enterobacteriaceae [START_REF] Cattaneo | Emerging resistant bacteria strains in bloodstream infections of acute leukaemia patients: results of a prospective study by the Rete Ematologica Lombarda (Rel)[END_REF] .

In order to prevent bacteria spreading to other patients, preventive measures are warranted, including patient isolation, limitations of transfer to other healthcare centers and management by dedicated staff, with consequent related increased healthcare costs, which are not easily affordable in most centers [START_REF] Birgand | Costs associated with implementation of a strict policy for controlling spread of highly resistant microorganisms in France[END_REF] . According to French recommendations [START_REF] Lepelletier | French recommendations for the prevention of " emerging extensively drug-resistant bacteria " ( eXDR )[END_REF] , for example, patients colonized with MDRB are not easily admitted in healthcare facilities not disposing of dedicated staff [START_REF] Lepelletier | French recommendations for the prevention of " emerging extensively drug-resistant bacteria " ( eXDR )[END_REF] .

New classes of antibiotics are under study to treat infections related to MDRB, and active research is ongoing to find effective decolonization strategies [START_REF] Bassetti | Management of KPC-producing Klebsiella pneumoniae infections[END_REF] . The use of oral gentamicin had been initially proposed in some MDR-gram negative strains, but failure is common, and the risk of selecting gentamicin-resistant strains may also be present [START_REF] Tascini | Oral gentamicin gut decontamination for prevention of KPC-producing Klebsiella pneumoniae infections: Relevance of concomitant systemic antibiotic therapy[END_REF][START_REF] Lübbert | Rapid emergence of secondary resistance to gentamicin and colistin following selective digestive decontamination in patients with KPC-2producing Klebsiella pneumoniae: A single-centre experience[END_REF] .

Fecal microbiota transplantation (FMT) is a procedure that has proven to be effective and safe in the treatment of recurrent Clostridium difficile infection (CDI), and it is now recommended as a therapeutic in this setting [START_REF] Rossen | Fecal microbiota transplantation as novel therapy in gastroenterology: A systematic review[END_REF] . Use of FMT in patients carrying MDRB is still investigational, but there are reports and case series showing its efficacy in this setting [START_REF] Manges | Fecal microbiota transplantation for the intestinal decolonization of extensively antimicrobial-resistant opportunistic pathogens: A review[END_REF][START_REF] Stalenhoef | Fecal microbiota transfer for multidrug-resistant gram-negatives: A clinical success combined with microbiological failure[END_REF] . Many concerns were initially raised about the feasibility of FMT in immunocompromised patients, such as those affected by hematologic malignancies, because of the theoretic potential for local and bloodstream infections. However, recent case reports revealed the efficacy and safety in this particular population [START_REF] Bilinski | Fecal Microbiota Transplantation Inhibits Multidrug-Resistant Gut Pathogens: Preliminary Report Performed in an Immunocompromised Host[END_REF][START_REF] Castro Cg De | Fecal microbiota transplant after hematopoietic SCT: report of a successful case[END_REF][START_REF] Kelly | Fecal Microbiota Transplant for Treatment of Clostridium diffi cile Infection in Immunocompromised Patients[END_REF] [START_REF] Bilinski | Fecal Microbiota Transplantation in Patients With Blood Disorders Inhibits Gut Colonization With Antibiotic-Resistant Bacteria: Results of a Prospective, Single-Center Study[END_REF] , including some of them with graft-versus-host disease (GVHD) after allogeneic hematopoietic stem cell transplantation (allo-HSCT).

In the current retrospective study, we report our experience with FMT in patients diagnosed with hematologic malignancies and undergoing FMT either before or after allo-HSCT.

Methods

In this single-center study, we retrospectively analyzed data on all consecutive adult patients diagnosed with hematologic malignancies who underwent FMT before or after allo-HSCT due to MDRB colonization. In our center microbiological screening is performed weekly in all inpatients, with consequent preventive measures in positive patients in order to limit MDRB spread, according to national guidelines [START_REF] Lepelletier | French recommendations for the prevention of " emerging extensively drug-resistant bacteria " ( eXDR )[END_REF] , as better detailed in online Supplemental material.

This study was approved by the Ethic Committee. The treatment plan was discussed in advance by a multidisciplinary team (hematologist, gastroenterologist, pharmacist) in order to approve the procedure. The decision was made on a patient to patient basis. All patients signed an informed consent mentioning the theoretical risks of the procedure, due to the actual investigational use of FMT in the field of MDRB and in patients with hematologic malignancies.

We considered eligibility to FMT in case of asymptomatic carriers or systemic infections from VRE, carbapenemase-producing Enterobacteriaceae (CPE) or CP-Pseudomonas aeruginosa. Rationales for FMT and MDRB decolonization were mainly to limit infectious complications related to these bacteria and to facilitate patients transfer in other departments such as intensive care units or rehabilitating centers.

Contemporary colonization from ESBL-producing bacteria was also registered in patients undergoing FMT. We therefore subsequently evaluated if FMT also allowed decolonization from these MDRB.

For the purpose of this retrospective analysis, we also classified MDRB as multi-drug (MDR), extensive-drug (XDR) and pan-drug-resistant (PDR) according to the definition proposed by Magiorakos et al. [START_REF] Magiorakos | bacteria: an international expert proposal for interim standard definitions for acquired resistance[END_REF] : MDR was defined as the presence of acquired non-susceptibility to at least one agent in three or more antimicrobial categories, XDR as non-susceptibility to at least one agent in all but two or fewer antimicrobial categories (i.e. bacterial isolates remain susceptible to only one or two categories) and PDR as non-susceptibility to all agents in all antimicrobial categories.. Details on donor selection, microbiological testing, fecal material preparation and delivery are available on online Supplemental material. Decolonization from VRE, CPE or CP-Pseudomonas aeruginosa after negative results on a minimum of three consecutive microbiological cultures (performed weekly) was defined as "major decolonization" while "persistent decolonization" was defined as the persistence of negative rectal swab until last follow-up after a first or second FMT, whenever this was feasible. In patients concomitantly colonized by ESBL-producing Enterobacteriaceae, "concomitant decolonization" was defined as negative results on at least three consecutive rectal swabs after FMT. The safety of the procedure was also registered. For all patients, data on significant infections, defined as bacteriemia or sepsis occurring during the first 90 days after FMT were also collected. Febrile neutropenia or fever of unknown origin was not considered as significant infectious episodes, but they were also recorded. In patients presenting either a relapse of MDRB colonization or experiencing FMT failure, a second attempt could be proposed.

Results

During the period between 2014 and 2017, 10 patients underwent FMT, 7 due to gut colonization without systemic infection by either CPE (Escherichia coli, n=1; Citrobacter freundii, n=2; Klebsiella pneumoniae, n=1), or CP-Pseudomonas aeruginosa (n=1) or VRE (n=2) and 3 after having experienced systemic infections from CP-Pseudomonas aeruginosa. The median age at FMT was 48 (range 16-64) years. Four patients underwent FMT as a decolonization strategy before allo-HSCT, with a median interval from FMT to transplant of 28 (range 9-46) days. Of note, one patient was contemporarily colonized by three different CPE. Two patients started conditioning regimen 3 days after FMT and the other two after a month. Six patients underwent FMT after allo-HSCT, with a median time from allo-HSCT to FMT of 163 (range 98-344) days. Of note, all patients undergoing FMT after allo-HSCT were still on immunosuppressive therapy at the time of FMT, with only one out of six presenting active grade IV steroid-dependent gut graft-versus-host disease (GVHD). Overall, six patients were also colonized by ESBL-producing Enterobacteriaceae. All ESBL-producing bacteria were classified as being MDR.

A frozen product was used in eight out of ten patients and enema was the preferred way of administration in all but one patient. This patient, indeed, presented a compromised neurological status due to a cerebral toxoplasmosis and she was not considered eligible for enema. Median quantity of donor stools was 84 g (range 43-104). At the time of FMT patients neutrophil count was > 1 x 10 9 /L in all patients but one that had a neutrophil count of 0.17 x 10^9/L (the one with steroidresistant GVHD). Platelet count was count > 20 x 10 9 /L in all patients.

Three patients required a second FMT: in one patient, after initial efficacy, VRE was again detectable 2 months after the first FMT. Of note, this patient developed multiple infectious episodes (particularly sinusitis and pneumonia), prompting to the frequent use of large spectrum antibiotics, thus probably leading to recurrence of VRE colonization. In the other two patients a second attempt was done due to the failure of the first procedure. In one patient this was mainly attributable to incorrect preparation with PEG (insufficient intake). After a second attempt with a correct preparation, indeed, VRE eradication was achieved and persisted until 20 months after FMT. At that time, indeed, VRE was detectable contemporarily to hematologic disease recurrence. In the last patient, first and second FMT mainly had a compassionate aim in order to treat active grade IV gut GVHD and contemporarily multiple infectious episodes rendering impossible antibiotics withdrawal, even during the 72 hours following FMT, as detailed below.

Globally, major decolonization (three consecutive negative microbiological cultures) was achieved in 7 out of 10 patients, including two patients after a second FMT (Figure 1). Persistent decolonization (negative microbiological cultures at last follow-up) was achieved in 6 out of ten patients after a median follow-up of 13 (range 4-40) months from FMT. As already mentioned, indeed, one patient presented a positive rectal swab for ERV 20 months after FMT meanwhile to disease relapse. She finally died due to hematological progression.

Failure occurred in the remaining three patients. The patient undergoing FMT with a compassionate aim had presented multiple infectious episodes from CP-Pseudomonas aeruginosa, rendering it impossible to stop antibiotics during the 72 hours after FMT. Moreover, grade IV gut GVHD was associated to intestinal occlusion, with need for an aspirating nasogastric tube, at time of FMT.

Despite two attempts with FMT, the procedure was a failure and the patient finally died. In the second patient, due to the difficulties encountered in the positioning of a nasogastric tube, FMT was administered by enema and the patient was not able to retain the product for the advised 2-3 hours. She then refused a second attempt. The third patient underwent FMT by enema from an unrelated donor and the hypothesis for FMT failure was that she received an insufficient quantity of stools (43 g), but what seems discordant with this hypothesis is that partial decolonization from concomitant ESBL-producing Enterobacteriaceae was achieved. A second attempt in this patient was not possible due to the unavailability of additional material.

Among the six patients concomitantly colonized from ESBL-producing Enterobacteriaceae, three obtained concomitant decolonization. Details on FMT performed before or after allo-HSCT are reported in Table 1. As an example of successful FMT, Figure 2 shows the case of the patient undergoing FMT from nasogastric tube, after experiencing breakthrough infectious episodes related to colonization from CP-Pseudomonas aeruginosa, with need for continuous in hospital care during the first year after allo-HSCT. After FMT, this patient did not experience any other infectious episode and outpatient care was finally possible.

According to the safety of FMT procedure, one patient presented constipation during the first 5 days after FMT which was favorably resolved after the use of laxatives, while two patients presented grade I diarrhea the day after FMT. No other major adverse events were observed.

Only one patient undergoing FMT before allo-HSCT developed a grade III acute gut graft-versus host disease at day +30 after allo-HSCT and at day +51 after FMT. Differential diagnosis with CMV colitis was evoked, and she favorably evolved after both antiviral and steroid treatment.

When looking at severe infectious episodes during the 90 days following FMT, in two of those patients undergoing FMT before allo-HSCT, documented bacteriemia without sepsis occurred early after allo-HSCT, favorably evolving after the introduction of large-spectrum antibiotics. In particular, one patient experienced a documented bacteriemia from multi-sensible Pseudomonas aeruginosa at day +80 after allo-HSCT while the other patient experienced a documented bacteriemia from an ESBL-producing Escherichia Coli at day 60 after allo-HSCT. The additional two patients undergoing FMT before allo-HSCT also received large spectrum antibiotic such as piperacillin-tazobactam or cephalosporins for febrile neutropenia without documentation. Interestingly, despite the use of large spectrum antibiotics, no cases of MDRB recurrence were observed in those four patients.

Of note, fungal and viral infections were observed in only one patient more than 6 months after FMT but these were not considered in relation to FMT because this patient was under systemic immunosuppressive treatments for a cortico-resistant extensive GVHD (lung, skin, mucosal) and infectious episodes exacerbated during immunosuppressive treatment. Among the other patients, neither fungal nor viral infections were observed.

Discussion

Increasing emergence and diffusion of MDRB represents a major public health problem, with higher mortality in patients experiencing infections, and high costs of prolonged in-hospital care and preventive measures used to limit diffusion to other patients [START_REF] Birgand | Costs associated with implementation of a strict policy for controlling spread of highly resistant microorganisms in France[END_REF][START_REF] Who | Antimicrobial resistance. Global Report on Surveillance[END_REF] .

Human gut microbiota, also named as "gut resistome", is the primary site for MDRB acquisition and colonization, being an important reservoir of antibiotic resistance genes [START_REF] Schaik | The human gut resistome[END_REF] . Patients diagnosed with hematological malignancies are at high risk of colonization from MDRB: conditioning regimens for allo-HSCT and intensive chemotherapy, indeed, significantly alter the gastrointestinal barrier and, subsequently, the composition of intestinal microbiota is largely modified. Moreover, patients affected by hematologic malignancies or undergoing allo-HSCT are at particular risk for MDRB colonization or infection due to the large, prolonged and, sometimes, improper use of large spectrum antibiotics [START_REF] Montassier | Chemotherapy-driven dysbiosis in the intestinal microbiome[END_REF] . Of note, most bloodstream infections in hematological patients derive from the gut, and infections are even more severe in those patients undergoing allo-HSCT, with high mortality rates of 36-95% [START_REF] Samet | Leukemia and risk of recurrent Escherichia coli bacteremia: Genotyping implicates E. Coli translocation from the colon to the bloodstream[END_REF][START_REF] Bilinski | Impact of Gut Colonization by Antibiotic-Resistant Bacteria on the Outcomes of Allogeneic Hematopoietic Stem Cell Transplantation: A Retrospective, Single-Center Study[END_REF] .

It has been largely reported that microbioma modifications are associated to worse survival, higher risk of infections and GVHD in patients undergoing allo-HSCT [START_REF] Taur | The effects of intestinal tract bacterial diversity on mortality following allogeneic hematopoietic stem cell transplantation[END_REF][START_REF] Jenq | Intestinal Blautia Is Associated with Reduced Death from Graft-versus-Host Disease[END_REF] . Therefore, efficacious decolonization strategies in this particular setting of patients are urgently needed.

Fecal microbiota transplantation is a fascinating decolonization strategy, that has been proven to be efficacious in patients with recurrent CDI [START_REF] Austin | Fecal microbiota transplantation in the treatment of clostridium difficile infections[END_REF][START_REF] Webb | Fecal microbiota transplantation for recurrent Clostridium difficile infection in hematopoietic stem cell transplant recipients[END_REF] . On the other hand, concerns were initially raised for the use of FMT as a decolonization strategy in immunocompromised patients, due to the possible risk of local or systemic infections after the inoculum of microbiota pathogens.

Recently, DeFilipp et al. investigated the use of third-party FMT with the use of oral capsules, as a strategy to restore microbioma diversity in patients undergoing allo-HSCT. They support the safety and feasibility of this procedure underlying the possibility that microbiome restauration early after allo-HSCT may be of benefit [START_REF] Defilipp | Third-party fecal microbiota transplantation following allo-HCT reconstitutes microbiome diversity[END_REF] .

Herein, we describe the results of FMT in 10 patients diagnosed with hematologic malignancies and undergoing FMT for MDRB colonization, namely CPE, CP-Pseudomonas aeruginosa or VRE, either before or after allo-HSCT. Decolonization was achieved in 7 out of 10 patients, this being persistent at last follow-up in 6 out 10 patients.

Our retrospective study not only suggests the efficacy of this procedure, but also its safety in patients with hematologic malignancies and undergoing allo-HSCT.

Of note, despite not being a selection criterion for FMT, we also registered patients concomitantly colonized from ESBL-producing enterobacteriaceae, with decolonization in 3 out of 6 cases.

We also showed that, in patients experiencing failure or relapse of MDRB colonization, a second FMT is feasible and efficacious. Interestingly, only three patients experienced significant infections after FMT.

Moreover, it is worth underlying the significant benefit of major decolonization in the patient who had experienced multiple infectious episodes due to a CP-Pseudomonas aeruginosa, limiting breakthrough infections.

Our results also highlight that despite administration of large spectrum antibiotics may hypothetically represent a risk of decolonization failure, the procedure remained effective in the majority of patients, without recurrence of MDRB in the majority of them despite use of broad spectrum antibiotics early after FMT.

Of note, in one patient VRE was detectable again at the time of disease relapse, despite no use of large-spectrum antibiotics just before this detection. One can speculate that disease relapse may have probably been associated to dysbiosis favoring selection of VRE, but conclusions cannot be drawn on one case.

Despite the initial aforementioned concerns in immunocompromised patients, results of FMT in this setting are promising in terms of both efficacy and safety [START_REF] Bilinski | Impact of Gut Colonization by Antibiotic-Resistant Bacteria on the Outcomes of Allogeneic Hematopoietic Stem Cell Transplantation: A Retrospective, Single-Center Study[END_REF][START_REF] Castro Cg De | Fecal microbiota transplant after hematopoietic SCT: report of a successful case[END_REF][START_REF] Kelly | Fecal Microbiota Transplant for Treatment of Clostridium diffi cile Infection in Immunocompromised Patients[END_REF] . A recent prospective study showed, indeed, that FMT allowed total eradication of MDRB in 60% of cases, without any significant adverse event after the procedure [START_REF] Bilinski | Fecal Microbiota Transplantation in Patients With Blood Disorders Inhibits Gut Colonization With Antibiotic-Resistant Bacteria: Results of a Prospective, Single-Center Study[END_REF] . The latter is the only prospective study published to date using FMT in 20 patients with blood disorders and colonized with MDRB. Differently from our series, in this study all types of MDRB were included and only a few patients underwent allo-HSCT.

In our Center, we only chose patients colonized with highly resistant bacteria and in particular those classified as eXDR according to French guidelines or those known to cause a significant higher risk of systemic infection with very poor prognosis (i.e. CP-Pseudomonas aeruginosa).

To date, there are no specific guidelines on the ideal timing, the best preparation of stools for FMT, and the best way of administration. In our experience, FMT was successfully undertaken either before or after allo-HSCT and, interestingly, it was also successful in two patients starting conditioning regimen for allo-HSCT 3 days after FMT.

As for stool preparation, frozen material was preferred in our Center particularly due to logistic reasons, although in two cases fresh stools were used, but this did not modify the results of FMT. It has recently been reported in a meta-analysis of patients receiving FMT for CDI, that the success rate of FMT was similar when using frozen or fresh stools [START_REF] Tang | Is frozen fecal microbiota transplantation as effective as fresh fecal microbiota transplantation in patients with recurrent or refractory Clostridium difficile infection: A meta-analysis?[END_REF] . Differently from most of the reported series of FMT for MDRB decolonization, we preferred enema as a way of administration, as this is associated with lower risk of inhalation as compared to nasogastric administration.

The mechanisms underlying the efficacy of FMT for MDRB decolonization are still not clear. In mice, Recent studies showed that recipient stool assumed donor-like taxonomic and functional composition immediately following FMT [START_REF] Moss | Long-term taxonomic and functional divergence from donor bacterial strains following fecal microbiota transplantation in immunocompromised patients[END_REF] . Therefore we hypothesize that FMT for MDRB decolonization works through the restoration of a more physiological microbiome thus increasing the ecological pressure on MDRB. However, given the absence of translational studies on antibiotic resistance genes and microbiota composition on patient's stool after FMT, we cannot exclude that FMT works through lowering of MDRB below the threshold of detection rather than through true elimination.

In our series, after FMT, almost all patients did not experience major infectious complications during the first 3 months after FMT and, of note, in those patients subsequently undergoing allo-HSCT, no severe infectious bacterial complications occurred during the early transplant phase.

Regarding the impact of FMT on GVHD, only one of our patients had a grade IV acute gut GVHD concomitant to a carbapenemase-producing Pseudomonas aeruginosa at the time of FMT. In this specific case, the procedure was not efficacious neither for MDRB nor for GVHD. However it is worth underlying that FMT was performed at a very late stage ("compassionate" use), that may also explain the failure of the procedure. Importantly, among the nine remaining patients, only one experienced grade III acute gut GVHD after FMT (with a possible differential diagnosis with CMV colitis). A role of FMT in causing GVHD in this patient cannot formally be excluded and this point may be addressed in a prospective clinical trial.

Early studies in mice and humans suggested a link between gut microbiota and propensity to GVHD, with mice treated with gut-decontaminating antibiotics developing GVHD less often [START_REF] Jones | Mortality and gross pathology of secondary disease in germfree mouse radiation chimeras[END_REF][START_REF] Shono | Increased GVHD-related mortality with broadspectrum antibiotic use after allogeneic hematopoietic stem cell transplantation in human patients and mice Yusuke[END_REF] . Recent results of a pilot study also highlight the possible advantage of microbiota modulation with FMT in patients affected by steroid-refractory or steroid-dependent GVHD [START_REF] Kakihana | Fecal microbiota transplantation for patients with steroid-resistant acute graft-versus-host disease of the gut[END_REF] .

With regards to donor choice, people living in the same household of the patient were preferred, when available, as they widely share the same pathogens and environment exposure, thus reducing the risk of transferring additional infectious agents from the donor to the recipient.

In line with previous reports, we consider that targeting gut microbiota in patients with impaired immune reconstitution in an attempt to reinstate a more equilibrated flora may favor stable eradication of the carrier status and prevent subsequent life threatening infections.

We are well aware of the limits of our study, being a retrospective one, including a low-number of patients, with non-homogeneous inclusion criteria and differences in FMT procedure according to patients, so that definitive conclusions cannot be drawn.

However, we consider that our results support the use of FMT as a promising strategy to manage the considerable potential risks associated with the MDRB carrier status in immunocompromised patients with intestinal dysbiosis and in those patients having experienced single or multiple systemic infections, with absence of breakthrough infections after decolonization and absence of MDRB recurrence despite the use of broad spectrum antibiotics in the majority of them. Furthermore, our results support again the safety of the procedure in this population, despite previous concerns in immunocompromised patients. These preliminary results underline the need for further prospective studies on the safety and efficacy of FMT.

Supplemental materials

Patient screening and preventive measures

In our center weekly screening for MDRB is performed in order to identify asymptomatic carriers with high risk of spreading MDRB to other patients. Screening modalities consist of weekly rectal swab. After MDRB identification, patients colonized with vancomycin-resistant (VRE) or carbapenemase-producing enterobacteriaceae (CPE) are cohorted and cared for by dedicated staff, as these two classes of bacteria are classified as emerging XDR (eXDR), i.e. bacteria that present an emerging infection control challenge widely in France. Of note, when those patients are candidates to rehabilitation centers before being discharged at home, they cannot be easily admitted to other healthcare facilities that often do not dispose of dedicated staff [START_REF] Lepelletier | French recommendations for the prevention of " emerging extensively drug-resistant bacteria " ( eXDR )[END_REF] .

Furthermore, in contact patients, defined as those patients having shared paramedical and/or medical healthcare workers with one or more patients colonized with VRE or CPE, cohorting is also warranted, with initial caring by another dedicated staff until three negative screening tests.

It is worth underlying that opportunistic saprophytic bacteria, such as CP-Pseudomonas aeruginosa,

have not been considered as eXDR in national guidelines. However, it has been already reported that patients experiencing systemic infections from CP-Pseudomonas aeruginosa have a high risk of death [START_REF] Cattaneo | Emerging resistant bacteria strains in bloodstream infections of acute leukaemia patients: results of a prospective study by the Rete Ematologica Lombarda (Rel)[END_REF] , and in our Center three consecutive patients (data not published) died during the aplastic phase of allo-HSCT due to bloodstream fatal infections from CP-Pseudomonas aeruginosa.

For these reasons, patients colonized with CP-Pseudomonas aeruginosa were considered at high risk of fatal complications and, despite not needing isolation and caring by dedicated staff, FMT was proposed to patients experiencing systemic infections or in those colonized in order to limit systemic infections.

A minimal platelet count of 20 x 10 9 /L was preferred in order to proceed to the FMT and use of platelet transfusion to reach that threshold before FMT was allowed.

Figure 1 .Figure 2 .

 12 Figure 1. Results of fecal microbiota transplantation

Figure

  Figure legend: CP, carbapenemase-producing; FMT, fecal microbiota transplantation; HSCT, hematopoietic stem cell transplantation.

  . Recently, Bilinski et al. reported the results of a prospective study evaluating FMT in 20 patients with MDRB gut colonization and contemporarily affected by hematologic malignancies. Overall 25 FMT were performed and 15/20 patients experienced complete MDRB decolonization

Table 1 .

 1 Characteristics of patients undergoing fecal microbiota transplantation before (a) or after (b) hematopoietic stem cell transplantation.

	a)
		1	2	3	4
	Patient sex	M	M	F	M
	Age at time of FMT, years	64	42	45	47
	Hematologic malignancy	AML	AML	AML	BPDCN
	Identified MDRB	CP-Pseudomonas aeruginosa	C P -Pseudomonas aeruginosa	CPE	CPE°
	Antimicrobial resistance category	XDR	MDR	MDR	MDR
	Concomitant MDR-ESBL-producing bacteria colonization, bacteria	Y	N	Y	N
	Systemic infections due to MDRB before FMT	Y	N	N	N
	Time from FMT to allo-HSCT (days)	41	46	16	9
	FMT donor	Daughter	Sister	Husband	Sister
	Way of administration	Enema	Enema	Enema	Enema
	Major decolonization	Y	Y	Y	Y
	Persistent decolonization	Y	Y	Y	Y
	Concomitant ESBL-producing bacteria decolonization	Y	N/A	N	N/A
	Follow-up after FMT, days	820	368	148	399
	Follow-up after allo-HSCT, days	779	322	132	390
	Status	Alive	Dead	Alive	Alive

Abbreviations: F, female; M, male; FMT, fecal microbiota transplantation; AML, acute myeloid leukemia; BPDCN, blastic plasmacytoid dendritic cell neoplasm; MDRB, multidrug-resistant bacteria; CP, carbapenemase-producing; CPE, carbapenemase-producing Enterobacteriaceae; XDR, extensively-drug resistant; MDR, multi-drug resistant; ESBL, extended-spectrum beta-lactamase; Y, yes, N, no; allo-HSCT, allogeneic hematopoietic stem cell transplantation; GVHD, graft-versus-host disease; N/A, not applicable; MPN, myeloproliferative neoplasm; ALL, acute lymphoblastic leukemia; VRE, vancomycin-resistant enterococci; PDR, pan-drug resistant.

Acknowledgments

The authors thank Prof. Junia V. Melo (University of Adelaide, Australia, and Imperial College, London) for medical editing of this manuscript. This study was supported by educational grants from the "Association for Training, Education and Research in Hematology, Immunology and Transplantation" (ATERHIT, Nantes, France).

Autorship contributions:

HS and MM designed the study. GB collected data. GB, FM, HS and MM wrote or revised the manuscript and all authors reviewed its final version.

JT was in charge of microbiological cultures. ACJ and MTB were in charge of product preparation. GB, FM, MTR, AR, ACM, EB, FG, RD were in charge of patients.

Conflict of interest statement: The authors declare no conflicts of interest in preparing this article.

Microbiological testing

For each patient, one rectal swab specimen was plated onto selective media: a screening medium designed to detect ESBL-producing enterobacteriaceae, ChromID ESBL (bioMérieux) and another designed to detect CP-bacteria, ChromID CARBA SMART (bioMérieux). A second rectal swab was used in an enrichment procedure, consisting of an overnight culture at 37°C in a specific broth before plating onto a screening medium designed to detect VRE, ChromID VRE (bioMérieux). All plates were incubated overnight at 37°C. Colonies growing on these selective media were identified at the species level by MALDI-TOF spectrometry. The production of ESBL was determined by an antibiogram and visualization of the characteristic "champagne cork" synergy between amoxicillin-clavulanate and third-generation cephalosporins disks. Carbapenemase production was determined by molecular analysis using the GeneXpert technology (Cepheid) and the Xpert Carba-R kit version 2 (detecting the most prevalent carbapenemases in France, OXA-48 and OXA-48-like enzymes, as well as NDM enzymes). Furthermore, VRE were also identified using the GeneXpert technology (Cepheid) and the Xpert VanA/VanB kit.

In patients achieving decolonization, rectal swabs and/or stool cultures were initially performed weekly and then at each follow-up visit. In patients considered as having achieved total and persistent decolonization, last follow-up for decolonization was considered as the date of the last available negative microbiological culture.

Patients and donors characteristics

current study was approved by the Ethic Committee. Each patient signed an informed consent mentioning all potential risks of the procedure as described in the paper. According to French regulations in such cases, each patient case was extensively discussed and approved as part of an "RCP" (Réunion de Concertation Pluridisciplinaire") which is a sort of large multidisciplinary meeting aimed to discuss difficult cases and approve unusual therapeutic procedures. The minutes and decisions of the RCP are recorded in writing, including the names of the participants and their feedback. Patients are informed about this discussion prior to signing the informed consent.

Large spectrum antibiotics were discontinued in the recipients 48-72 hours prior to the procedure and, when possible, use of antibiotics was avoided during at least 72 hours after the procedure.

Stools were preferentially obtained from healthy related or unrelated donors. Of note, related donors not necessarily coincided with allo-HSCT donors. According to regulatory recommendations, potential donors were selected after a previous questionnaire. Donor age was preferentially between 18 and 65 years. Excluded were people who had presented digestive disorders (i.e. diarrhea) within the 3 months prior to donation or having a chronic disease and/or chronic treatments, cases with antibiotic intake within 3 months before the donation, people having been living in the tropics during the three months prior to donation or having been hospitalized abroad for more than 24 hours in the 12 months prior to donation. History of typhoid fever was also considered as exclusion criteria. In people fulfilling inclusion criteria, a complete biological and microbiological assessment was then performed including: serology for Treponema pallidum, human immunodeficiency virus, Human T-Lymphotropic Virus, Hepatitis A, B and C, cytomegalovirus, Epstein-Barr virus, amebiasis, Strongyloides strecoralis ; stool examination for standard culture, Clostridium difficile, multi-resistant bacteria, norovirus, Cryposporidium, parasites. If the biological and microbiological panel was negative, a minimum of 50 g of stools were collected.

Fecal material, prepared as described below, was delivered either by enema or via nasogastric tube.

A bowel preparation was performed the day before the FMT by administration of 4 liters of polyethylene glycol (PEG) based solution. For nasogastric administration, patients had to fast for at least 12 hours before transplantation and they received proton pump inhibitors the day before and the morning of the FMT. In the case of enema administration, patients were asked to retain the product for at least 2-3 hours.

Product preparation

Transplants were prepared in the Saint Antoine Hospital pharmacy. In case of freezing, the stool preparation is usually performed in two steps. In the first, preparation and freezing, the stools are manipulated in an extractor hood dedicated to this activity, in the 6 hours following emission. A total of 50-100g stools are weighted and mixed with a sterile cryopreservative saline solution (300mL glycerol+ saline solution 0.9% 10/90 V/V) using sterile blender, containers and medical devices (syringes, filters). The suspension is filtrated through sterile gauze compresses mounted in a funnel to remove solid residues, before freezing at -80°C. If in screening tests an exclusion criterion is fulfilled, the suspension is destroyed. The second step of the preparation procedure starts the day before FMT, when the frozen microbiota solution is placed in a refrigerator (between 4 and 8°C) for an overnight thawing. The thawed suspension is then transferred either to an enema bag (lower gastro intestinal tract delivery) to which 200mL of sterile saline solution are added, or to 50-mL syringes (colonoscopy or nasoduodenal delivery) as ready to be used. . On the other hand, when FMT is performed with fresh stools, fecal materials need to be prepared the day of FMT within the 6 hours following stools emission. In this case stool preparation is performed in a single step, without freezing.

Safety testing of the fecal product was done according to French recommendations 1 .


Michael Von Cotta-Schönberg

Silvio Enea

; Piccolomini

Sylvius Aeneas

Silvius Aeneas

Pope Pius II Piccolomini

Papa

Michael V Cotta-Schönberg

Oration "Prius sanctissime praesul" of Enea

In the beginning of 1445, Enea Silvio Piccolomini, secretary in the Roman Chancery of Emperor Friedrich III, arrived at the papal court in Rome on a mission from the emperor. The mission was part of ongoing negotiations between the imperial court and the papal court aiming at ending the German state of Neutrality between Pope Eugenius IV and the Council of Basel, and at Germany returning to the obedience of the Roman pope. Formerly in his career, Piccolomini had been a fervent conciliarist and had written and spoken much against the pope. During his years in Austria/Germany from 1442 and onwards, however, he had slowly migrated to a papalist position, and he

(Collected Orations of Pope Pius II; 7) Foreword

In 2007, I undertook a project of publishing the Latin texts with English translations of the orations of Enea Silvio Piccolomini / Pope Pius II. Altogether 80 1 orations (including papal responses to ambassadorial addresses) are extant today, though more may still be held, unrecognized, in libraries and archives.

At a later stage the project was expanded to include ambassadors' orations to the pope, of which about 40 are presently known.

I do not, actually, plan to publish further versions of the present volume, but I do reserve the option in case I -during my future studies -come across other manuscripts containing interesting versions of the oration or if important new research data on the subject matter are published, making it appropriate to modify or expand the present text. It will therefore always be useful to check if a later version than the one the reader may have previously found via the Internet is available.

I shall much appreciate to be notified by readers who discover errors and problems in the text and translation or unrecognized quotations.

7 September 2019 MCS 1 81 orations, if the "Cum animadverto" is counted is a Piccolomini-oration, see oration "Quam laetus" [18], Appendix An important factor in this development was the position of his mentor and superior, Chancellor Heinrich Schlick who for various reasons -some quite opportunistic -sponsored the papal cause and the cessation of the German Neutrality 3 between the Council of Basel 4 and Pope Eugenius IV.

The development is documented in the oration "Si putarem" [5], held by the chancellor at the imperial court on 4 April 1444 and mostly written by Piccolomini.

In the beginning of 1445, Piccolomini was sent by the emperor to Pope Eugenius to present an imperial proposal for a new council with the object of restoring church unity. It had not been planned that way: the emperor had wanted his Chancellor Schlick to undertake this mission, but shortly before the chancellor's projected departure court affairs prevented his travelling to Rome.

In his stead the emperor sent one of his secretaries, the chancellor's close collaborator and protégé, Enea Silvio Piccolomini, thus providing Piccolomini with an unexpected opportunity for reconciliation with the pope and a boost of his career in the clerical direction. 5

The mission was not without some risk to Piccolomini himself, since his conversion to the papal cause was not yet public, [START_REF] Ady | Pius II (AEneas Silvius Piccolomini) -the Humanist Pope[END_REF] and since he had, in 1435, been personally involved in a Milanese plot 1 CO, I, 13 (Meserve, I, pp. 52-59); Ady, pp. 87-89; Boulting, pp. 154-160; Koller, p. 105; Mitchell, pp. 98-100; O'Brien, p. 113; Pastor, II, pp. 258-259; Pückert, p. 237; Stiebert, pp. 272-275 He did so first very consciously and deliberately as a prominent member of the council of Basel, and, in particular as a publicist who introduced powerful new weapons into the assembly's anti-papal assaults. As imperial secretary, legate, and adviser, he continued to take positions on conciliar authority and on the role of general councils that were perceived in the curia as hostile to papal sovereignty (O'Brien, p. 113). Piccolomini's importance to the conciliar movement and the Papacys's perception of it, however, appear not to be well documented. Moreover, his change to the papalist cause actually occurred only a couple of years after his joining the imperial court -and had been privately communicated to Piccolomini's connections at the papal court to seize the pope's person. 1 It was therefore necessary for him to formally submit to the pope and to obtain his forgiveness for his former conciliarist activities at the Council of Basel.

When he was received by Eugenius, he therefore first asked for the pope's forgiveness which he received, and only afterwards did he undertake his negotiations on behalf of the emperor

In his Commentarii, Pius wrote about the mission:

After this, the Council of Basel rejected the proposed terms of peace, and Though many learned men had assembled there [in Basel], he was the only one 4 to have praised the majesty and the acts of the council in his writings. For that reason he knew Eugenius to be his enemy. But after the dissolution of the council5 he came to the pope and presented a splendid apology, which is now read by many, showing so great virtue and uncommon gracefulness that the past was forgotten and he rose greatly in the estimation of the pope and the other fathers. [START_REF] Ady | Pius II (AEneas Silvius Piccolomini) -the Humanist Pope[END_REF] Of course the pope forgave the man kneeling before him, 1 the emperor's personal envoy, and though it was delicately put, he now expected Piccolomini to act in the interests of the Papacy at the imperial court:

You are now in a position to defend the truth and be of service to the Church. Hereafter we shall forget past wrongs, and as long as you do well, we shall love you well. 2 Having become reconciled with the pope and concluded his mission to the papal court, Piccolomini returned to the emperor. The imperial mission actually failed since the pope -welladvisedly -later rejected the emperor's proposal for another council, but his personal mission, the reconciliation with the pope, was a success: it marked the end of his conversion from the conciliarist to the papalist cause and gave the first serious impetus to his future ecclesiastic career. 3

Themes

The theme of the oration was Piccolomini's confession of his conciliarist errors:

While in Basel, I said, and wrote, and did much against you: I do not deny it. But my intention was not to harm you, but to benefit the Church of God. Working against you, I thought that I was serving God. I erred: who will deny it? [Sect. 1]

Piccolomini's errors were not in themselves terrible, since so many others had held them, too, and had afterwards been happily reconciled with the pope. Piccolomini carefully mentions some of them, among whom was an outstanding member of the College of Cardinals, a luminary of the papal court, Cardinal Cesarini who as papal legate had represented the pope in the crusade ending disastrously at Varna the year before, and where Cesarini himself was killed, a martyr for the Faith. Aggravating circumstances, however, would have been Piccolomini's fervent writings and orations in the conciliarist cause, his contributions to the election of the antipope and his administration, and the time it had taken for him to reach his present papalist convictions.

He defended himself quite reasonably, however:

I admit that when I perceived the error of the Basileans, I did not immediately hasten to your side, as many did. Fearing to abandon one error just to fall into another -as often happens when those who wish to avoid Charybdis fall into Scylla -I went to those who were considered neutral, so as not to pass from one extreme to another without a pause for reflexion. So I have stayed with the emperor for three years 2 where I have heard more and more about the conflicts between the Basileans and your legates. I no longer have any doubts that the truth is with you. [Sect. 2]

Date, place, format and audience

The exact date of Piccolomini's audience with the pope, where the oration was delivered, is not known, but it was sometime in February or March 1445. According to Pückert, Piccolomini was in Rome already before 16 January. 3 The place was the Apostolic Palace in Rome, and the oration was delivered before Pope Eugenius and cardinals Landriano and Le Jeune. The format is that of an oration, though a very short one. In his Commentarii, Pius does not use the term "oration", but says: "jussus fuisset dicere … inquit …". However, a margin note in the Reginensis Latinus 1995 (see below) says: "Oratio Enee ad pontificem". And Platina calls it an oration. 1 Though the context of the speech was semiprivate and the audience very small (but august), the occasion was a formal one and for Piccolomini personally of the greatest importance. So, he had undoubtedly prepared his text very, very carefully.

Text 2

Though the oration had circulated individually and was much read, 3 it was not included in the Collected Orations of Pius II, compiled under his direct supervision in 1462.

It was, however, included in his Commentarii, together with a number of orations given by the pope during the last years of his pontificate (1462-1464). 4 It is not known if or to what extent it was revised in this connection.

Manuscripts

Manuscripts containing the individually circulated oration are not presently known.

The oration is only extant in the manuscripts containing the Commentarii, of which the two primary ones are: (1423-1429) when Piccolomini studied at the same university [START_REF] Ady | Pius II (AEneas Silvius Piccolomini) -the Humanist Pope[END_REF] Ludovico Pontano (1409-1439): Italian lawman. Pronotary of the papal Curia. One of the representatives of Alfonso V at the Council of Basel. Friend of Piccolomini who admired him greatly [START_REF]Sigla and abbreviations R = Roma / Biblioteca Apostolica Vaticana / Reginensis Latinus[END_REF] During that period the University of Paris was staunchly conciliarist 8 "gymnasia" [2] I admit that when I perceived the error of the Basileans, I did not immediately hasten to your side, as many did. Fearing to abandon one error just to fall into another -as often happens when those who wish to avoid Charybdis fall into Scylla 1 -I went to those who were considered neutral, so as not to pass from one extreme to another without a pause for reflexion. So I have stayed with the emperor for three years 2 where I have heard more and more about the conflicts between the Basileans and your legates. I no longer have any doubts that the truth is with you. Therefore, when the emperor 3 wished to send me to Your Clemency, I was very happy to obey, believing that in this this way I could gain your favour. Now I stand before you, and having sinned out of ignorance, I beg you to forgive me.

 Roma / Biblioteca
Afterwards I shall set forth the emperor's matter.

Table of volumes in Collected Orations of Pope Pius II. 12 vols. Edited and translated by Michael von Cotta-Schönberg. Table of contents

  When, in 1442, Enea Silvio Piccolomini left the service of antipope Felix V to become a secretary in the Imperial Chancery of Friedrich III, he was still a fervent conciliarist, believing in the supremacy of the council over the pope and supporting the, by now, schismatic Council of Basel and its antipope against Pope Eugenius IV, 2 deposed by the Council of Basel in 1439. Over the next two years, however, a number of events made him abandon the conciliarist cause and join the cause of the Pope Eugenius.

	I. INTRODUCTION 1. Context 1
	I.	INTRODUCTION
		1.	Context
		2.	Themes
		3.	Date, place, format and audience
		4.	Text
			4.1. Manuscripts
			4.2. Editions
			4.3. Present edition
		5.	Sources
		6.	Bibliography
		7.	Sigla and abbreviations
	II. TEXT AND TRANSLATION

  Aeneas was sent to confer with Eugenius. When he reached Siena, however his relations all tried to deter him from going to Rome because he had been opposed to Eugenius at Basel. Eugenius, they said, remembered nothing so long as an injury and was both vindictive and cruel. Aeneas replied His second mission was to Eugenius. His friends advised against it since he had offended Eugenius gravely, when his dignity was under continuous attack in Basel. But he became reconciled with Eugenius and when his mission was accomplished he returned … 1His second contemporary biographer, Bartolomeo Platina, wrote:When Eugenius and the emperor began to negotiate on ending the schism, Enea was sent to Rome in this matter. When he came to Siena his friends urged him not to go to Eugenius, fearing that he would detest Enea because he had often attacked his authority, in epistles and orations, at the Council of Basel. But being courageous and confident in his own innocence, he disregarded the pleas of his relations, and continued to Rome. First he excused himself in an elegant oration before Eugenius for having followed the authority of the men who had secured the approval of the Council of Basel.2 Then he began to negotiate with him on the matters concerning which he had been sent by the emperor, though many were against it.

that he could not imagine the emperor's ambassador would be in danger in Rome and that he must either perform the duty he had undertaken or die in the attempt. And so, ignoring the protests of his weeping family, he continued on his way. An old intimate of his, Gerardo Landriani, cardinal of Como, an honest priest and faithful friend, introduced him to the presence of Eugenius. I should like to quote here the words Aeneas addressed to the pope, for they are brief and worth recording. Two cardinals, Como and Amiens, were standing by the pope. First, they invoked apostolic authority to absolve Aeneas of all the blame he had incurred by taking the side of the Council of Basel. Then Aeneas came before Eugenius. He was given permission to kiss his foot, hand and cheek; he presented his credentials and was asked to speak. [Here follows the text of the oration.] Eugenius replied: "We know that you have sinned grievously against us, but since you confess your error we must pardon you. The Church is a loving mother, who never remits the due penalty if a son denies his sin, nor insists on it when he confesses. Now you know the truth. See that you do not let it go. Strive by good works to regain the divine grace which you lost by wicked acts. You are now in a position to defend the truth and be of service to the Church. Hereafter we shall forget past wrongs and, so long as you do well, we shall love you well." After this they proceeded to speak of ecclesiastical matters, about which Eugenius wished time to meditate. ….When he heard from Eugenius that he would send an ambassador to the emperor, he went back to Siena.

2 3

Pius' contemporary biographer, Gianantonio Campano wrote:

1

Voigt, I, p. 80 2 1 April 1445 3 CO, I, 13 (Meserve, I, pp. 52-59) 3

And his friend, Lodrisio Crivelli, wrote in his De expeditione Pii II:

  Cesarini had been a friend and mentor of Piccolomini's at the Council of Basel, before he returned to the pope in 1438,1 and Piccolomini was indeed well-advised to mention him in his apology:But I did so together with many and great men. I followed Giuliano, Cardinal of Sant'Angelo, Archbishop Niccolò of Palermo, and Ludovico Pontano, a notary of you own See, men who were considered the eyes of the law and teachers of truth. And what should I say about the University of Paris and the other universities in the world, many of which opposed you? Who might not have erred together with so great men?[Sect. 2] Piccolomini might also have mentioned Cardinal Cusa, another luminary of the papal court, who after an early conciliarist phase left the Council of Basel in 1436, to be appointed cardinal by Eugenius in 1446.

	1 Frizzi, p. 72: la propria difesa, che ad Eugenio recitò il Piccolomini, ne desarmò l'íra per quanto aveva detto e scritto
	contro di lui nel Concilio di Basilea 2 CO, I, 13 (Meserve, I, p. 56-57) 3 Baldi, pp. 102-103: L'immagine, tramandata dai Commentarii, del Piccolomini che, inginocchiato ai piedi di Eugenio
	IV, chiede al pontifice il perdono per il suo passato conciliare, ha in effetti assunto un'importanza centrale nella
	riconstruzione dello svolgimento della vicenda piccolominiana e proprio la decisione di prestare obbedienza ad Eugenio

IV è stata anzi considerata spesso come il punto di arrivo di un processo di maturazione, di "svolta", che si afferma progressivamente e che lo porta a riavvicinarsi al papato romano. L'episodio, in particolare, aprirebbe al Piccolomini nuove, importante possibilità di carriera, di azione; contribuirebbe alla sua ascesa, non solo nella cancelleria, ma anche, entro il volgere di breve tempo, nella gerarchia ecclesiastica

  For the textual transmission of Pius II's orations, see Collected Orations of Pope Pius II, vol. 1, ch. 5 3 See Crivelli above 4 See Collected Orations of Pope Pius II, vol. 1, sect. 5.1.4[1] Holy Bishop, before I deliver the message 1 from the emperor, I shall say a little about myself. I know that you have been told many things about me which are neither good nor worthy of mention, and those who denounced me to you did not lie. While in Basel, 2 I said, and wrote and did much against you: I do not deny it. But my intention was not to harm you, but to benefit the Church of God. Working against you, I thought that I was serving God. 3 I erred: who will deny it? But I did so together with many and great men. I followed Giuliano, Cardinal of Sant'Angelo, 4 Archbishop Niccolò of Palermo, 5 and Ludovico Pontano, 6 a notary of you own See, men who were considered the eyes of the law and teachers of truth. And what should I say about the University of Paris 7 and the other universities 8 in the world, many of which opposed you? Who might not have erred together with so great men? Aeneis, 7.267; 11.1762 Piccolomini arrived in Basel in 1432, as secretary to Cardinal Capranica. He was often absent from Basel, however, on extended missions to Milan and Scotland. Returning to Basel, he became a functionary of the council in 1436 and remained so even after the Council suspended the pope in 1438, at which point the remaining papalists left the council and returned to the papal court. The rump council went on to elect a new pope, Felix V, in 1439, whose secretary Piccolomini became. In 1442 he joined the imperial court of Friedrich III which still officially favoured the Neutrality between Pope Eugenius IV and the Council in Basel, declared by the German nation in 1438 3 John, 16, 2 4 Cesarini, Giuliano (1398-1444): created cardinal by Martinus V in 1426. Died at the Battle of Varna, 1444. Mentor and friend of Piccolomini. When he arrived, as papal legate, to take up the presidency of the Council of Basel in 1431, Cardinal Cesarini was a firm conciliarist, and he actively and successfully resisted the pope's attempt to speedily dissolve the Council. As the Council's opposition to the pope became more and more pronounced, not to say rabid, Cesarini gradually lost faith in it and finally left, in January 1438, in order to rejoin the pope and become one of his most trusted advisors 5 Niccolò Tedeschi (1386-1445): canon lawyer and abbot of Benedictine monastery in Catania; Archbishop of Palermo from 1435; created cardinal by the antipope, Felix V, in 1440. Called "lucerna juris". Taught civil law at the University of Siena from 1418-1430, including during the period

	Apostolica Vaticana
	Reginensis Latinus 1995, ff. 15r-15v
	 Roma / Biblioteca dell'Accademia dei Lincei
	Corsinianus 147, ff. 11v
	1 Zimolo, p. 101: se … eleganti oratione purgat 2

1 Vergilius:

1

  Otto, p. 82. The Latin proverb was coined by Gualtier de Lille: Alexandreis, 5, 301 2 Since 1442 3 Friedrich III (Habsburg) (1415-1493): Duke of Austria (as Friedrich V) from 1424. Elected King of Germany and Holy Roman Emperor in 1440, crowned emperor in Rome in 1452

Zimolo, p. 14: Altera [legatione] ad Eugenium, adversantibus amicis, quod offendisse illum graviter videbatur, oppugnata totiens ad Basileam eius dignitate. Eo reconciliato et rebus ex sentientia confectis, divertit …

Primarily Cardinal Cesarini

Zimolo, p. 100: Agi de abolendo scismate ab Eugenio et Caesare interim est coeptum. Eius rei causa Aeneas Romam missus, cum Saenis iter faceret, a suis vehementer rogatur ne ad Eugenium proficisceretur: veriti enim sunt ne in eum pontifex graviter animadverteret, quod eius auctoritas in concilio basiliensi ab Aenea epistolis et orationibus persaepe impugnata fuerat. At homo fortissimus sua innocentia fretus, spretis cognatorum precibus, Romam contendit, seque primum ad Eugenium eleganti oratione purgat, quod eorum auctoritatem secutus esset, quorum sententia basileense concilium fuerat approbatum. Deinde de his rebus cum eo agere coepit, quarum gratia ab imperatore venerat, cui certe studio partium multi sunt adversati

Piccolomini was certainly not the only one to have praised the council in words and writings

If Crivelli's refers to the pope's translation of the Council of Basel to Ferrara in 1438, eight years had actually passed before Piccolomini was reconciled with the pope

Crivelli, p. 38: …cum multi doctissimi viri eo convenissent, solus tamen illius synodi majestatem et res gestas scriptis suis illustravit. Quorum gratia cum se Eugenio infensum sciret, post concilii tamen dissolutionem ad Pontificem veniens et luculentissimo, qui nunc a multis legitur, usus apologetico vis tantae virtutis et non vulgaris gratiae dignitatisque locum, obliterata praeteritorum memoria, apud eum ceterosque patres invenit

Becker,

Since 1442

Pückert, p. 238. Baldi, p. 102, gives the time of Piccolomini's arrival in Rome as March. Voigt, II, p. 340, only says that Piccolomini began his travel to Rome in the beginning of the year

Though the two manuscripts have been very ably edited by van Heck, Totaro and others, they have -as a matter of principle -been collated directly for the present edition

For an analysis of Piccolomini's use of sources, see Collected Orations of Pope PiusII, ch. 8

http://reader.digitale-sammlungen.de/de/fs1/object/display/bsb10016831_00005.html

Editions

The oration has been published in the editions of the Commentarii, e.g.

Present edition

For principles of edition (incl. orthography) and translation, see Collected Orations of Pope Pius II, vol. 1, ch. 9-10.

Text:

The edition is based on the Reginensis and the Corsinianus. 1 The lead text is from the Corsinianus, with variants from the Reginensis in the critical apparatus.

Pagination:

Pagination is from the Reginensis.

Abbreviations CO = Pius II: Commentarii rerum memorabilium quae suis temporibus contigerunt [1464] II. TEXT AND TRANSLATION