Robotically assisted deployment of flow diverter stents for the treatment of cerebral and cervical aneurysms.

BACKGROUND: Despite the growing sophistication of robot-assisted surgery, it is necessary to demonstrate that robots can reliably perform complex procedures on site and then remotely. Although a flow diverter stent is one of the most effective and widely used devices, its placement is sometimes challenging. OBJECTIVE: To evaluate the feasibility and safety of the CorPath GRX robotic platform for the embolization of cerebral and cervical aneurysms using flow diverter stents. METHODS: We performed a single-center technical study of the first 10 flow diverter stent deployments with the CorPath GRX Robotic System (Corindus Inc, Waltham, Massachusetts, USA) for the treatment of cerebral aneurysms between April and October 2022. RESULTS: Ten patients underwent robot-assisted embolization with flow diverter stents: there were nine intracranial aneurysms (paraclinoid n=6; posterior communicating artery aneurysm n=1; anterior communicating artery n=2) and one cervical aneurysm. Four procedures were performed with coils plus a flow diverter stent, one was performed with woven endobridge plus a flow diverter stent and four were performed with flow diverter stents alone. Of these procedures, two were performed with telescoping flow diverters.All flow diverter stents were deployed with robotic assistance, with only one partial conversion to a manual technique (caused by guidewire torquability limitations). No perioperative complications were observed. CONCLUSION: Robot-assisted flow diverter stent deployment using the CorPath GRX platform is feasible and appears to be safe. Larger, in-depth studies of the technique's safety and benefits are now warranted.

Optimizing coronary artery opacification and 3D reconstruction from human cadaver hearts in anatomy research.

OBJECTIVE: This study seeks to identify the ideal dilution rate of a radiopaque product to optimize the visualization of coronary arteries and their branches within human cadaver hearts. The process involves obtaining images in the anatomy laboratory and subsequently constructing a three-dimensional model. MATERIALS AND METHODS: We utilized 30 human hearts fixed in 10 % formalin (9 females and 21 males) with a mean age of 79 ± 5 years. The initial experiment, involving the first four hearts (referred to as "group 1"), encountered difficulties in opacifying coronary arteries. In this phase, a probabilistic injection of 20 % Visipaque and 80 % latex, with coronary sinus ostium closure, was performed. The optimal mixture ratio was then determined as 33 % Visipaque and 66 % latex. Recognizing the need for on-site injection at the CT Scan table, this protocol was applied to the subsequent 11 hearts in "group 2." Closure of the coronary sinus was deemed unnecessary. The final 15 hearts, constituting "group 3," revealed that the injection should be gradual, maintaining controlled pressure between 120 and 150 mm Hg. Post-injection, hearts were scanned with the injected coronary arteries using an Optima 660 CT scanner. Two-dimensional images were acquired with parameters set at 64 × 0.625 mm, 100 kV, 300-400 mA, and a rotation of 0.5 s. Subsequently, 3D reconstruction was conducted using Advantage Workstation 4.7 (GE Healthcare) and volume rendering with Volume Viewer software, version 15. RESULTS: Significant differences in the percentage of opacified coronaries were observed among the three groups (p < 0.005). This variation underscores the learning curve and comprehension required before establishing a reliable method. Group 1 (N = 4) demonstrated minimal opacification, group 2 (N = 11) displayed partial opacification, while group 3 (N = 15) achieved 100 % opacification of coronary arteries. CONCLUSION: The successive experiments culminated in the development of a protocol for CT imaging, enabling accurate three-dimensional reconstruction of the normal anatomy of the main and secondary coronary arteries. Our work is grounded in a series of progressively refined and successful experiments.

Clostridioides difficile MreE (PBP2) variants facilitate clinical disease during cephalosporin exposures.

Cephalosporins are the most common triggers of healthcare-associated Clostridioides difficile infections (CDI). Here, we confirm gene-level drivers of cephalosporin resistance and their roles in promoting disease. Genomic-epidemiologic analyses of 306 C. difficile isolates from a hospital surveillance program monitoring asymptomatic carriers and CDI patients identified prevalent third-generation cephalosporin resistance to ceftriaxone at >256 ug/mL in 26% of isolates. Resistance was associated with patient cephalosporin exposures 8-10 days before C. difficile detection. Genomic analyses identified variants in the mreE penicillin binding protein 2 (PBP2) associated with resistance to multiple beta-lactam classes. Transfer of variants into susceptible strain CD630 elevated resistance to first and third-generation cephalosporins. Transfer into the mouse-infective strain ATCC 43255 enabled disease when mice were exposed to 500ug/mL cefoperazone, a dose that inhibited the isogenic susceptible strain. Our findings establish roles of cephalosporins and mreE-cephalosporin-resistant variants in CDI and provide testable genetic loci for detecting resistance in patient strains.

Hydrodynamic and Hemodynamic Interactions in Chronic Hydrocephalus.

BACKGROUND: During a cardiac cycle, intracranial pressure is related to arterial entry into the cranium and its interaction with intracranial compliance. The arterial inflow is compensated by intracranial compliance and, initially, the flushing of cerebrospinal fluid (CSF) into the cervical subarachnoid spaces. Our objective is to analyze the interactions between intracranial arteriovenous exchange and cerebrospinal fluid oscillations. METHOD: A total of 23 patients (73 ± 8 years) with suspected chronic hydrocephalus (CH) underwent an infusion test and phase-contrast MRI. Rout is an important factor in the diagnosis of CH. Patients were divided into 2 populations: probableCH (Rout: resistance to CSF outflow) (Rout > 12 mmHg/mL/min, 13 patients) and unlikelyCH (Rout < 12 mmHg/mL/min, 10 patients). We measured the intracranial vascular volume (arteriovenous stroke volume: SVvasc) and CSF (CSF stroke volume at upper cervical level: SVCSF) volume variations during the cardiac cycle. RESULTS: In the whole population, we observed a significant correlation between SVvasc and SVCSF (R2 = 0.43; p = 0.0007). In the population unlikelyCH, this correlation was significant (R2 = 0.76; p = 0.001). In the population probableCH, this correlation was not significant (R2 = 0.17, p = 0.16). CONCLUSIONS: These results show that the link between the compliance of the oscillating CSF and the abrupt arterial inflow seems to be altered in CH. CSF oscillations between intracranial and cervical fluid spaces limit the impact of the abrupt arterial inflow.

HRMAS 13C NMR and genome-scale metabolic modeling identify threonine as a preferred dual redox substrate for Clostridioides difficile.

Stickland-fermenting Clostridia preferentially ferment amino acids to generate energy and anabolic substrates for growth. In gut ecosystems, these species prefer dual redox substrates, particularly mucin-abundant leucine. Here, we establish how theronine, a more prevalent, mucinabundant substrate, supports dual redox metabolism in the pathogen Clostridioides difficile. Realtime, High-Resolution Magic Angle Spinning NMR spectroscopy, with dynamic flux balance analyses, inferred dynamic recruitment of four distinct threonine fermentation pathways, including ones with intermediate accrual that supported changing cellular needs for energy, redox metabolism, nitrogen cycling, and growth. Model predictions with 13C isotopomer analyses of [U-13C]threonine metabolites inferred threonine's reduction to butyrate through the reductive leucine pathway, a finding confirmed by deletion of the hadA 2-hydroxyisocaproate CoA transferase. In vivo metabolomic and metatranscriptomic analyses illustrate how threonine metabolism in C. difficile and the protective commensal Paraclostridium bifermentans impacts pathogen colonization and growth, expanding the range of dual-redox substrates that modulate host risks for disease.

Hemodynamic and Hydrodynamic Pathophysiology in Chiari Type 1 Malformations: Towards Understanding the Genesis of Syrinx.

BACKGROUND: The pathophysiology of this association of type 1 Chiari malformation (CM1) and syrinxes is still unknown. There is an alteration in the dynamics of neurofluids (cerebrospinal fluid, arterial and venous blood) during the cardiac cycle in CM1. Our objective is to quantify CSF or arterial blood or venous blood flow in patients with Chiari syndrome (CS) with and without syrinxes using phase-contrast MRI (PCMRI). METHODS: We included 28 patients with CM1 (9 with syrinxes, 19 without). Morphological MRI with complementary PCMRI sequences was performed. We analyzed intraventricular CSF, subarachnoid spaces CSF, blood, and tonsillar pulsatility. RESULTS: There is a highly significant correlation (p < 0.001) between cerebral blood flow, cerebral vascular expansion volume and venous drainage distribution. Venous drainage distribution is significantly inversely correlated with oscillatory CSF volume at the level of the foramen magnum plane [-0.37 (0.04)] and not significantly correlated at the C2C3 level [-0.37 (0.05)] over our entire population. This correlation maintained the same trend in patients with syrinxes [-0.80 (<0.01)] and disappeared in patients without a syrinx [-0.05 (0.81)]. CONCLUSION: The distribution of venous drainage is an important factor in intracranial homeostasis. Impaired venous drainage would lead to greater involvement of the CSF in compensating for arterial blood influx, thus contributing to syrinx genesis.

Anatomical and biometric study of the radial artery on human cadavers dissected in the anatomy laboratory.

OBJECTIVE: To study the anatomy and biometry of the radial artery and to report the implications of this study for daily practice in Cardiac Surgery. METHOD: Radial arteries from 45 human cadavers (28 males and 17 females, average age 79.2 (92) fixed in 10% formalin were dissected. The proximal and distal internal calibers and lengths of these radial arteries were measured. RESULTS: Our results showed the presence of a single radial artery variation in 2.3% of the entire sample (1.1% of the 90 dissected upper limbs). The distance between the epicondyle and the emergence of the radial artery was 32.4 (6.67) mm in men and 30.7 (9.00) mm in women, with an average of 31.8 (7.58) mm. For the right upper limb, the mean proximal internal caliber of the radial artery was 3.16 (0.56) mm and its mean distal internal caliber was 2.62 (0.66) mm. For the left upper limb, the mean proximal internal caliber of the radial artery was 3.17 (0.59) mm and its mean distal internal caliber was 2.64 (0.68) mm. The mean length of the left radial artery was 197.0 (17) mm. The mean length of the right radial artery was 201.0 (33) mm. CONCLUSION: It is very important to be aware of the possible anatomical variation of the radial artery. Despite its rarity, this knowledge may ensure a better safety and reliability of the harvesting technique for use as a graft.

Central in vivo mechanisms by which C. difficile's proline reductase drives efficient metabolism, growth, and toxin production.

Clostridioides difficile (CD) is a sporulating and toxin-producing nosocomial pathogen that opportunistically infects the gut, particularly in patients with depleted microbiota after antibiotic exposure. Metabolically, CD rapidly generates energy and substrates for growth from Stickland fermentations of amino acids, with proline being a preferred reductive substrate. To investigate the in vivo effects of reductive proline metabolism on C. difficile's virulence in an enriched gut nutrient environment, we evaluated wild-type and isogenic ΔprdB strains of ATCC43255 on pathogen behaviors and host outcomes in highly susceptible gnotobiotic mice. Mice infected with the ΔprdB mutant demonstrated extended survival via delayed colonization, growth and toxin production but ultimately succumbed to disease. In vivo transcriptomic analyses demonstrated how the absence of proline reductase activity more broadly disrupted the pathogen's metabolism including failure to recruit oxidative Stickland pathways, ornithine transformations to alanine, and additional pathways generating growth-promoting substrates, contributing to delayed growth, sporulation, and toxin production. Our findings illustrate the central role for proline reductase metabolism to support early stages of C. difficile colonization and subsequent impact on the pathogen's ability to rapidly expand and cause disease.

Phenotypes controlled by the Brucella abortus two component system BvrR/BvrS are differentially impacted by BvrR phosphorylation.

Brucella abortus is a zoonotic pathogen whose virulence depends on its ability to survive intracellularly at the endoplasmic reticulum derived compartment. The two-component system BvrR/BvrS (BvrRS) is essential for intracellular survival due to the transcriptional control of the type IV secretion system VirB and its transcriptional regulator VjbR. It is a master regulator of several traits including membrane homeostasis by controlling gene expression of membrane components, such as Omp25. BvrR phosphorylation is related to DNA binding at target regions, thereby repressing or activating gene transcription. To understand the role of BvrR phosphorylation we generated dominant positive and negative versions of this response regulator, mimicking phosphorylated and non-phosphorylated BvrR states and, in addition to the wild-type version, these variants were introduced in a BvrR negative background. We then characterized BvrRS-controlled phenotypes and assessed the expression of proteins regulated by the system. We found two regulatory patterns exerted by BvrR. The first pattern was represented by resistance to polymyxin and expression of Omp25 (membrane conformation) which were restored to normal levels by the dominant positive and the wild-type version, but not the dominant negative BvrR. The second pattern was represented by intracellular survival and expression of VjbR and VirB (virulence) which were, again, complemented by the wild-type and the dominant positive variants of BvrR but were also significantly restored by complementation with the dominant negative BvrR. These results indicate a differential transcriptional response of the genes controlled to the phosphorylation status of BvrR and suggest that unphosphorylated BvrR binds and impacts the expression of a subset of genes. We confirmed this hypothesis by showing that the dominant negative BvrR did not interact with the omp25 promoter whereas it could interact with vjbR promoter. Furthermore, a global transcriptional analysis revealed that a subset of genes responds to the presence of the dominant negative BvrR. Thus, BvrR possesses diverse strategies to exert transcriptional control on the genes it regulates and, consequently, impacting on the phenotypes controlled by this response regulator.

Anatomical and biometric study of the mitral valve apparatus: application in valve repair surgery.

OBJECTIVE: Most mitral valve repair techniques provide excellent surgical results by removing regurgitation, but all of these techniques simultaneously reduce posterior valve mobility. A comprehensive biometric study of the mitral valve apparatus will provide landmarks that would help improve this posterior valve mobility. MATERIALS AND METHODS: Thirty one (31) human hearts have been studied, from 14 women and 17 men. The characteristics of the studied sample were analyzed descriptively. The difference in means of the variables between women and men were tested using a Student t test. Correlations between the different measures were determined by simple regression analysis. Mean values are shown with ± 1 standard deviation and the limit of significance was set at 0.05. RESULTS: The mean weight of the hearts was 275.3 ± 2.4 g. The anteroposterior diameter of the mitral annulus was 29.3 ± 1.22 mm, the intertrigonal distance was 25.2 ± 3.50 mm and the anterior leaflet to posterior leaflet ratio was 1.9 ± 0.10, the length of the chordae A2 = 19.4 ± 1.15 mm and P2 = 14.5 ± 0.85 mm. The length of the anterior papillary muscle averaged 30.9 ± 7.20 mm and that of the posterior one 30.0 ± 8.75 mm. The comparison of the different values measured between women and men showed no statistically significant difference (p > 0.05). There was no correlation between these different measured values (p > 0.05). CONCLUSION: A perfect knowledge of anatomy and biometry is therefore essential to offer alternative techniques that reproduce the real anatomy and physiology with a complete reconstruction of the mitral valve.

Elucidating dynamic anaerobe metabolism with HRMAS 13C NMR and genome-scale modeling.

Anaerobic microbial metabolism drives critical functions within global ecosystems, host-microbiota interactions, and industrial applications, yet remains ill-defined. Here we advance a versatile approach to elaborate cellular metabolism in obligate anaerobes using the pathogen Clostridioides difficile, an amino acid and carbohydrate-fermenting Clostridia. High-resolution magic angle spinning nuclear magnetic resonance (NMR) spectroscopy of C. difficile, grown with fermentable 13C substrates, informed dynamic flux balance analysis (dFBA) of the pathogen's genome-scale metabolism. Analyses identified dynamic recruitment of oxidative and supporting reductive pathways, with integration of high-flux amino acid and glycolytic metabolism at alanine's biosynthesis to support efficient energy generation, nitrogen handling and biomass generation. Model predictions informed an approach leveraging the sensitivity of 13C NMR spectroscopy to simultaneously track cellular carbon and nitrogen flow from [U-13C]glucose and [15N]leucine, confirming the formation of [13C,15N]alanine. Findings identify metabolic strategies used by C. difficile to support its rapid colonization and expansion in gut ecosystems.

Incidental discovery of a sacral rib: a case report.

PURPOSE: The observation of an accessory bone joined to the sacrum (suggestive of a sacral rib) prompted us to describe the structure's morphology and its anatomic relationships and to consider its morphogenesis and clinical implications. METHOD: A 38-year-old woman underwent computed tomography, to characterize the extension of a thoracic mass. We compared our observations with the literature data. RESULTS: We observed a voluminous accessory bone located behind and to the right of the sacrum. The bone was articulated with the third sacral vertebra and featured a head and three processes. These characteristics were suggestive of a sacral rib. We also observed involution of the gluteus maximus. CONCLUSION: This accessory bone probably resulted from overdevelopment of a costal process and an absence of fusion with the primitive vertebral body. Sacral ribs are rare and usually asymptomatic but appear to be more prevalent in young women. The adjacent muscles are often abnormal. Awareness of the potential presence of this bone is essential for surgeons who operate on the lumbosacral junction.

Impact of Shunt Placement on CSF Dynamics.

BACKGROUND: CSF dynamics are disturbed in chronic hydrocephalus (NPH). We hypothesise that these alterations reflect a disturbance of intracranial compliance. The aim of our study is to investigate the variations in intracranial hydrodynamics in NPH after ventricular shunt surgery. PATIENTS AND METHOD: We included 14 patients with definite NPH. All patients improved after ventriculoperitoneal shunting. The patients underwent an analysis of intracranial haemodynamics by phase-contrast MRI (pcMRI) preoperatively, at 6 months postoperatively, and at 1 year postoperatively. We analysed the dynamics of intraventricular CSF at the level of the aqueduct of Sylvius (SVAQU) and CSF at the level of the high cervical subarachnoid spaces (SVCERV). We calculated the ratio between SVAQU and SVCERV, called CSFRATIO, which reflects the participation of intraventricular pulsatility in overall intracranial CSF pulsatility. RESULTS: SVAQU significantly (p = 0.003) decreased from 240 ± 114 µL/cc to 214 ± 157 µL/cc 6 months after shunt placement. Six months after shunt placement, SVCERV significantly (p = 0.007) decreased from 627 ± 229 µL/cc to 557 ± 234 µL/cc. Twelve months after shunt placement, SVCERV continued to significantly (p = 0.001) decrease to 496 ± 234 µL/cc. CSFRATIO was not changed by surgery. CONCLUSIONS: CSF dynamics are altered by shunt placement and might be a useful marker of the shunt's effectiveness-especially if pressure values start to rise again. The detection of changes in CSF dynamics would require a reference postoperative pcMRI measurement for each patient.

c-di-AMP signaling is required for bile salt resistance, osmotolerance, and long-term host colonization by Clostridioides difficile.

To colonize the host and cause disease, the human enteropathogen Clostridioides difficile must sense, respond, and adapt to the harsh environment of the gastrointestinal tract. We showed that the production and degradation of cyclic diadenosine monophosphate (c-di-AMP) were necessary during different phases of C. difficile growth, environmental adaptation, and infection. The production of this nucleotide second messenger was essential for growth because it controlled the uptake of potassium and also contributed to biofilm formation and cell wall homeostasis, whereas its degradation was required for osmotolerance and resistance to detergents and bile salts. The c-di-AMP binding transcription factor BusR repressed the expression of genes encoding the compatible solute transporter BusAA-AB. Compared with the parental strain, a mutant lacking BusR was more resistant to hyperosmotic and bile salt stresses, whereas a mutant lacking BusAA was more susceptible. A short exposure of C. difficile cells to bile salts decreased intracellular c-di-AMP concentrations, suggesting that changes in membrane properties induce alterations in the intracellular c-di-AMP concentration. A C. difficile strain that could not degrade c-di-AMP failed to persist in a mouse gut colonization model as long as the wild-type strain did. Thus, the production and degradation of c-di-AMP in C. difficile have pleiotropic effects, including the control of osmolyte uptake to confer osmotolerance and bile salt resistance, and its degradation is important for host colonization.

Insights on the Hydrodynamics of Chiari Malformation.

BACKGROUND: We propose that the appearance of a ptosis of the cerebellar tonsils and syringomyelia is linked to its own hemohydrodynamic mechanisms. We aimed to quantify cerebrospinal fluid (CSF) and cerebral blood flow to highlight how neurofluid is affected by Chiari malformations type 1(CMI) and its surgery. METHODS: We retrospectively included 21 adult patients with CMI who underwent pre- and postoperative phase contrast MRI (PCMRI) during the period from 2001 to 2017. We analyzed intraventricular CSF, subarachnoid spaces CSF, blood, and tonsils pulsatilities. RESULTS: In preoperative period, jugular venous drainage seems to be less preponderant in patients with syringomyelia than other patients (venous correction factor: 1.49 ± 0.4 vs. 1.19 ± 0.1, p = 0.05). After surgery, tonsils pulsatility decreased significantly (323 ± 175 µL/cardiac cycle (CC) vs. 194 ± 130 µL/CC, p = 0.008) and subarachnoid CSF pulsatility at the foramen magnum increased significantly (201 ± 124 µL/CC vs. 363 ± 231 µL/CC, p = 0.02). After surgery, we found a decrease in venous flow amplitude (5578 ± 2469 mm3/s vs. 4576 ± 2084 mm3/s, p = 0.008) and venous correction factor (1.98 ± 0.3 vs. 1.20 ± 0.3 mm3/s, p = 0.004). CONCLUSIONS: Phase-contrast MRI could be a useful additional tool for postoperative evaluation and follow-up, and is complementary to morphological imaging.

Predictive regulatory and metabolic network models for systems analysis of Clostridioides difficile.

We present predictive models for comprehensive systems analysis of Clostridioides difficile, the etiology of pseudomembranous colitis. By leveraging 151 published transcriptomes, we generated an EGRIN model that organizes 90% of C. difficile genes into a transcriptional regulatory network of 297 co-regulated modules, implicating genes in sporulation, carbohydrate transport, and metabolism. By advancing a metabolic model through addition and curation of metabolic reactions including nutrient uptake, we discovered 14 amino acids, diverse carbohydrates, and 10 metabolic genes as essential for C. difficile growth in the intestinal environment. Finally, we developed a PRIME model to uncover how EGRIN-inferred combinatorial gene regulation by transcription factors, such as CcpA and CodY, modulates essential metabolic processes to enable C. difficile growth relative to commensal colonization. The C. difficile interactive web portal provides access to these model resources to support collaborative systems-level studies of context-specific virulence mechanisms in C. difficile.

In vivo commensal control of Clostridioides difficile virulence.

Leveraging systems biology approaches, we illustrate how metabolically distinct species of Clostridia protect against or worsen Clostridioides difficile infection in mice by modulating the pathogen's colonization, growth, and virulence to impact host survival. Gnotobiotic mice colonized with the amino acid fermenter Paraclostridium bifermentans survive infection with reduced disease severity, while mice colonized with the butyrate-producer, Clostridium sardiniense, succumb more rapidly. Systematic in vivo analyses revealed how each commensal alters the gut-nutrient environment to modulate the pathogen's metabolism, gene regulatory networks, and toxin production. Oral administration of P. bifermentans rescues conventional, clindamycin-treated mice from lethal C. difficile infection in a manner similar to that of monocolonized animals, thereby supporting the therapeutic potential of this commensal species. Our findings lay the foundation for mechanistically informed therapies to counter C. difficile disease using systems biology approaches to define host-commensal-pathogen interactions in vivo.

Ser/Thr Kinase-Dependent Phosphorylation of the Peptidoglycan Hydrolase CwlA Controls Its Export and Modulates Cell Division in Clostridioides difficile.

Cell growth and division require a balance between synthesis and hydrolysis of the peptidoglycan (PG). Inhibition of PG synthesis or uncontrolled PG hydrolysis can be lethal for the cells, making it imperative to control peptidoglycan hydrolase (PGH) activity. The synthesis or activity of several key enzymes along the PG biosynthetic pathway is controlled by the Hanks-type serine/threonine kinases (STKs). In Gram-positive bacteria, inactivation of genes encoding STKs is associated with a range of phenotypes, including cell division defects and changes in cell wall metabolism, but only a few kinase substrates and associated mechanisms have been identified. We previously demonstrated that STK-PrkC plays an important role in cell division, cell wall metabolism, and resistance to antimicrobial compounds in the human enteropathogen Clostridioides difficile In this work, we characterized a PG hydrolase, CwlA, which belongs to the NlpC/P60 family of endopeptidases and hydrolyses cross-linked PG between daughter cells to allow cell separation. We identified CwlA as the first PrkC substrate in C. difficile We demonstrated that PrkC-dependent phosphorylation inhibits CwlA export, thereby controlling hydrolytic activity in the cell wall. High levels of CwlA at the cell surface led to cell elongation, whereas low levels caused cell separation defects. Thus, we provided evidence that the STK signaling pathway regulates PGH homeostasis to precisely control PG hydrolysis during cell division.IMPORTANCE Bacterial cells are encased in a PG exoskeleton that helps to maintain cell shape and confers physical protection. To allow bacterial growth and cell separation, PG needs to be continuously remodeled by hydrolytic enzymes that cleave PG at critical sites. How these enzymes are regulated remains poorly understood. We identify a new PG hydrolase involved in cell division, CwlA, in the enteropathogen C. difficile Lack or accumulation of CwlA at the bacterial surface is responsible for a division defect, while its accumulation in the absence of PrkC also increases susceptibility to antimicrobial compounds targeting the cell wall. CwlA is a substrate of the kinase PrkC in C. difficile PrkC-dependent phosphorylation controls the export of CwlA, modulating its levels and, consequently, its activity in the cell wall. This work provides a novel regulatory mechanism by STK in tightly controlling protein export.

Varicocele complicating an anterior lumbar interbody fusion: a case report.

Anterior lumbar interbody fusion (ALIF) is performed for the surgical management of lumbar degenerative disc disease with excellent results, particularly for discogenic low back pain. Commonly reported complications associated with this approach include vessel injury, retrograde ejaculation, and ureteral and viscus organ injury. The development of a varicocele after ALIF has not been previously described in the literature. We report a case of varicocele in a 35-year-old patient who underwent ALIF via a left retroperitoneal approach. No intraoperative complications were identified. The postoperative course was uneventful. He was discharged from the hospital on the 5th postoperative day. Three months after surgery, he complained of discomfort and scrotal pain. Examination revealed a grade 3 varicocele according to the Dubin and Amelar classification. Scrotal Doppler US demonstrated dilatation of the veins of the pampiniform plexus. A lumbar CT scan revealed a bulky left spermatic vein closed to the ureter. The patient was treated with platelet anti-aggregation. He was seen at control intervals of 1, 3 and 5 months. Progress was seen as we had a regression of clinical signs. Varicocele appears as an uncommon complication of ALIF. After reviewing the literature, we describe the occurrence of a varicocele following ALIF, its pathophysiology, and its treatment options.

Identification of RNAs bound by Hfq reveals widespread RNA partners and a sporulation regulator in the human pathogen Clostridioides difficile.

Noncoding RNAs (ncRNA) have emerged as important components of regulatory networks governing bacterial physiology and virulence. Previous deep-sequencing analysis identified a large diversity of ncRNAs in the human enteropathogen Clostridioides (Clostridium) difficile. Some of them are trans-encoded RNAs that could require the RNA chaperone protein Hfq for their action. Recent analysis suggested a pleiotropic role of Hfq in C. difficile with the most pronounced effect on sporulation, a key process during the infectious cycle of this pathogen. However, a global view of RNAs interacting with C. difficile Hfq is missing. In the present study, we performed RNA immunoprecipitation high-throughput sequencing (RIP-Seq) to identify Hfq-associated RNAs in C. difficile. Our work revealed a large set of Hfq-interacting mRNAs and ncRNAs, including mRNA leaders and coding regions, known and potential new ncRNAs. In addition to trans-encoded RNAs, new categories of Hfq ligands were found including cis-antisense RNAs, riboswitches and CRISPR RNAs. ncRNA-mRNA and ncRNA-ncRNA pairings were postulated through computational predictions. Investigation of one of the Hfq-associated ncRNAs, RCd1, suggests that this RNA contributes to the control of late stages of sporulation in C. difficile. Altogether, these data provide essential molecular basis for further studies of post-transcriptional regulatory network in this enteropathogen.