The GPI-anchor biosynthesis pathway is critical for syncytiotrophoblast differentiation and placental development.

The glycosylphosphatidylinositol (GPI) biosynthetic pathway in the endoplasmic reticulum (ER) is crucial for generating GPI-anchored proteins (GPI-APs), which are translocated to the cell surface and play a vital role in cell signaling and adhesion. This study focuses on two integral components of the GPI pathway, the PIGL and PIGF proteins, and their significance in trophoblast biology. We show that GPI pathway mutations impact on placental development impairing the differentiation of the syncytiotrophoblast (SynT), and especially the SynT-II layer, which is essential for the establishment of the definitive nutrient exchange area within the placental labyrinth. CRISPR/Cas9 knockout of Pigl and Pigf in mouse trophoblast stem cells (mTSCs) confirms the role of these GPI enzymes in syncytiotrophoblast differentiation. Mechanistically, impaired GPI-AP generation induces an excessive unfolded protein response (UPR) in the ER in mTSCs growing in stem cell conditions, akin to what is observed in human preeclampsia. Upon differentiation, the impairment of the GPI pathway hinders the induction of WNT signaling for early SynT-II development. Remarkably, the transcriptomic profile of Pigl- and Pigf-deficient cells separates human patient placental samples into preeclampsia and control groups, suggesting an involvement of Pigl and Pigf in establishing a preeclamptic gene signature. Our study unveils the pivotal role of GPI biosynthesis in early placentation and uncovers a new preeclampsia gene expression profile associated with mutations in the GPI biosynthesis pathway, providing novel molecular insights into placental development with implications for enhanced patient stratification and timely interventions.

Complex agricultural landscapes host more biodiversity than simple ones: A global meta-analysis.

Managing agricultural landscapes to support biodiversity conservation requires profound structural changes worldwide. Often, discussions are centered on management at the field level. However, a wide and growing body of evidence calls for zooming out and targeting agricultural policies, research, and interventions at the landscape level to halt and reverse the decline in biodiversity, increase biodiversity-mediated ecosystem services in agricultural landscapes, and improve the resilience and adaptability of these ecosystems. We conducted the most comprehensive assessment to date on landscape complexity effects on nondomesticated terrestrial biodiversity through a meta-analysis of 1,134 effect sizes from 157 peer-reviewed articles. Increasing landscape complexity through changes in composition, configuration, or heterogeneity significatively and positively affects biodiversity. More complex landscapes host more biodiversity (richness, abundance, and evenness) with potential benefits to sustainable agricultural production and conservation, and effects are likely underestimated. The few articles that assessed the combined contribution of linear (e.g., hedgerows) and areal (e.g., woodlots) elements resulted in a near-doubling of the effect sizes (i.e., biodiversity level) compared to the dominant number of studies measuring these elements separately. Similarly, positive effects on biodiversity are stronger in articles monitoring biodiversity for at least 2 y compared to the dominant 1-y monitoring efforts. Besides, positive and stronger effects exist when monitoring occurs in nonoverlapping landscapes, highlighting the need for long-term and robustly designed monitoring efforts. Living in harmony with nature will require shifting paradigms toward valuing and promoting multifunctional agriculture at the farm and landscape levels with a research agenda that untangles complex agricultural landscapes' contributions to people and nature under current and future conditions.

Acetic acid stress response of the acidophilic sulfate reducer Acididesulfobacillus acetoxydans.

Acid mine drainage (AMD) waters are a severe environmental threat, due to their high metal content and low pH (pH <3). Current technologies treating AMD utilize neutrophilic sulfate-reducing microorganisms (SRMs), but acidophilic SRM could offer advantages. As AMDs are low in organics these processes require electron donor addition, which is often incompletely oxidized into organic acids (e.g., acetic acid). At low pH, acetic acid is undissociated and toxic to microorganisms. We investigated the stress response of the acetotrophic Acididesulfobacillus acetoxydans to acetic acid. A. acetoxydans was cultivated in bioreactors at pH 5.0 (optimum). For stress experiments, triplicate reactors were spiked until 7.5 mM of acetic acid and compared with (non-spiked) triplicate reactors for physiological, transcriptomic, and membrane lipid changes. After acetic acid spiking, the optical density initially dropped, followed by an adaptation phase during which growth resumed at a lower growth rate. Transcriptome analysis revealed a downregulation of genes involved in glutamate and aspartate synthesis following spiking. Membrane lipid analysis revealed a decrease in iso and anteiso fatty acid relative abundance; and an increase of acetyl-CoA as a fatty acid precursor. These adaptations allow A. acetoxydans to detoxify acetic acid, creating milder conditions for other microorganisms in AMD environments.

Intracranial self-stimulation reverses impaired spatial learning and regulates serum microRNA levels in a streptozotocin-induced rat model of Alzheimer disease.

BACKGROUND: The assessment of deep brain stimulation (DBS) as a therapeutic alternative for treating Alzheimer disease (AD) is ongoing. We aimed to determine the effects of intracranial self-stimulation at the medial forebrain bundle (MFB-ICSS) on spatial memory, neurodegeneration, and serum expression of microRNAs (miRNAs) in a rat model of sporadic AD created by injection of streptozotocin. We hypothesized that MFB-ICSS would reverse the behavioural effects of streptozotocin and modulate hippocampal neuronal density and serum levels of the miRNAs. METHODS: We performed Morris water maze and light-dark transition tests. Levels of various proteins, specifically amyloid-ß precurser protein (APP), phosphorylated tau protein (pTAU), and sirtuin 1 (SIRT1), and neurodegeneration were analyzed by Western blot and Nissl staining, respectively. Serum miRNA expression was measured by reverse transcription polymerase chain reaction. RESULTS: Male rats that received streptozotocin had increased hippocampal levels of pTAU S202/T205, APP, and SIRT1 proteins; increased neurodegeneration in the CA1, dentate gyrus (DG), and dorsal tenia tecta; and worse performance in the Morris water maze task. No differences were observed in miRNAs, except for miR-181c and miR-let-7b. After MFB-ICSS, neuronal density in the CA1 and DG regions and levels of miR-181c in streptozotocin-treated and control rats were similar. Rats that received streptozotocin and underwent MFB-ICSS also showed lower levels of miR-let-7b and better spatial learning than rats that received streptozotocin without MFB-ICSS. LIMITATIONS: The reversal by MFB-ICSS of deficits induced by streptozotocin was fairly modest. CONCLUSION: Spatial memory performance, hippocampal neurodegeneration, and serum levels of miR-let-7b and miR-181c were affected by MFB-ICSS under AD-like conditions. Our results validate the MFB as a potential target for DBS and lend support to the use of specific miRNAs as promising biomarkers of the effectiveness of DBS in combatting AD-associated cognitive deficits.

Alcohol dehydrogenase system acts as the sole pathway for methanol oxidation in Desulfofundulus kuznetsovii strain TPOSR.

Desulfofundulus kuznetsovii is a thermophilic, spore-forming sulphate-reducing bacterium in the family Peptococcaceae. In this study, we describe a newly isolated strain of D. kuznetsovii, strain TPOSR, and compare its metabolism to the type strain D. kuznetsovii 17T. Both strains grow on a large variety of alcohols, such as methanol, ethanol and propane-diols, coupled to the reduction of sulphate. Strain 17T metabolizes methanol via two routes, one involving a cobalt-dependent methyl transferase and the other using a cobalt-independent alcohol dehydrogenase. However, strain TPOSR, which shares 97% average nucleotide identity with D. kuznetsovii strain 17T, lacks several genes from the methyl transferase operon found in strain 17T. The gene encoding the catalytically active methyl transferase subunit B is missing, indicating that strain TPOSR utilizes the alcohol dehydrogenase pathway exclusively. Both strains grew with methanol during cobalt starvation, but growth was impaired. Strain 17T was more sensitive to cobalt deficiency, due to the repression of its methyl transferase system. Our findings shed light on the metabolic diversity of D. kuznetsovii and their metabolic differences of encoding one or two routes for the conversion of methanol.

Sex and Gender-related Disparities in Clinical Characteristics and Outcomes in Heart Transplantation.

PURPOSE OF REVIEW: Limited research has been conducted on sex disparities in heart transplant (HT). The aim of this review is to analyse the available evidence on the influence of sex and gender-related determinants in the entire HT process, as well as to identify areas for further investigation. RECENT FINDINGS: Although women make up half of the population affected by heart failure and related mortality, they account for less than a third of HT recipients. Reasons for this inequality include differences in disease course, psychosocial factors, concerns about allosensitisation, and selection or referral bias in female patients. Women are more often listed for HT due to non-ischaemic cardiomyopathy and have a lower burden of cardiovascular risk factors. Although long-term prognosis appears to be similar for both sexes, there are significant disparities in post-HT morbidity and causes of mortality (noting a higher incidence of rejection in women and of malignancy and cardiac allograft vasculopathy in men). Additional research is required to gain a better understanding of the reasons behind gender disparities in eligibility and outcomes following HT. This would enable the fair allocation of resources and enhance patient care.

Coupled C, H, N, S and Fe biogeochemical cycles operating in the continental deep subsurface of the Iberian Pyrite Belt.

Microbial activity is a major contributor to the biogeochemical cycles that make up the life support system of planet Earth. A 613 m deep geomicrobiological perforation and a systematic multi-analytical characterization revealed an unexpected diversity associated with the rock matrix microbiome that operates in the subsurface of the Iberian Pyrite Belt (IPB). Members of 1 class and 16 genera were deemed the most representative microorganisms of the IPB deep subsurface and selected for a deeper analysis. The use of fluorescence in situ hybridization allowed not only the identification of microorganisms but also the detection of novel activities in the subsurface such as anaerobic ammonium oxidation (ANAMMOX) and anaerobic methane oxidation, the co-occurrence of microorganisms able to maintain complementary metabolic activities and the existence of biofilms. The use of enrichment cultures sensed the presence of five different complementary metabolic activities along the length of the borehole and isolated 29 bacterial species. Genomic analysis of nine isolates identified the genes involved in the complete operation of the light-independent coupled C, H, N, S and Fe biogeochemical cycles. This study revealed the importance of nitrate reduction microorganisms in the oxidation of iron in the anoxic conditions existing in the subsurface of the IPB.

Determinants of activity and efficacy of anti-PD1/PD-L1 therapy in patients with advanced solid tumors recruited in a clinical trials unit: a longitudinal prospective biomarker-based study.

Immune-checkpoint inhibitors (ICI) have revolutionized the therapeutic landscape of cancer. However, optimal patient selection is still an unmet need. One-hundred-forty-six patients with metastatic cancer candidates to ICI at the Hospital Clinic of Barcelona Clinical Trials Unit were prospectively recruited in this observational study. Blood samples were collected at different timepoints, baseline LIPI score calculated and pre-ICI archived tissues retrieved to evaluate PD-L1, tumor-infiltrating lymphocytes (TILs) and PD1 mRNA levels. Tumor assessments were centrally reviewed by RECIST 1.1 criteria. Associations with overall response rates (ORR), durable clinical benefit (DCB), progression-free survival (PFS) and overall survival (OS) were performed with univariable/multivariable logistic and Cox regressions, where appropriate. At a median follow-up of 26.9 months, median PFS and OS were 2.7 and 12.9 months. Response rates were 17.8% with duration of response (DOR) of 4.4 months. LIPI score was independently associated with PFS (p = 0.025) and OS (p < 0.001). Immunotherapy-naïve status was independently associated with better PFS (p = 0.005). Time-to-best response (TTBR) and ORR (p < 0.001 both) were associated with better OS at univariate analysis. PFS and DOR were moderately correlated with OS (p < 0.001 both). A PD-L1 10% cut-off detected worse/best responders in terms of ORR (univariate p = 0.011, multivariate p = 0.028) and DCB (univariate p = 0.043). PD1 mRNA levels were strikingly associated to complete responses (p = 0.021). To resume, in our prospective observational pan-cancer study, baseline LIPI score, immunotherapy-naïve status, cancer type and RT before starting ICI were the most relevant clinical factors independently correlated with immunotherapy outcomes. Longer TTBR seemed to associate with better survival, while PD1 mRNA and PD-L1 protein levels might be tumor-agnostic predictive factors of response to ICI and should be furtherly explored.

Detoxification of methylglyoxal by the glyoxalase system is required for glutathione availability and virulence activation in Listeria monocytogenes.

Listeria monocytogenes is a Gram-positive, food-borne pathogen that lives a biphasic lifestyle, cycling between the environment and as a facultative intracellular pathogen of mammals. Upon entry into host cells, L. monocytogenes upregulates expression of glutathione synthase (GshF) and its product, glutathione (GSH), which is an allosteric activator of the master virulence regulator PrfA. Although gshF mutants are highly attenuated for virulence in mice and form very small plaques in host cell monolayers, these virulence defects can be fully rescued by mutations that lock PrfA in its active conformation, referred to as PrfA*. While PrfA activation can be recapitulated in vitro by the addition of reducing agents, the precise biological cue(s) experienced by L. monocytogenes that lead to PrfA activation are not known. Here we performed a genetic screen to identify additional small-plaque mutants that were rescued by PrfA* and identified gloA, which encodes glyoxalase A, a component of a GSH-dependent methylglyoxal (MG) detoxification system. MG is a toxic byproduct of metabolism produced by both the host and pathogen, which if accumulated, causes DNA damage and protein glycation. As a facultative intracellular pathogen, L. monocytogenes must protect itself from MG produced by its own metabolic processes and that of its host. We report that gloA mutants grow normally in broth, are sensitive to exogenous MG and severely attenuated upon IV infection in mice, but are fully rescued for virulence in a PrfA* background. We demonstrate that transcriptional activation of gshF increased upon MG challenge in vitro, and while this resulted in higher levels of GSH for wild-type L. monocytogenes, the glyoxalase mutants had decreased levels of GSH, presumably due to the accumulation of the GSH-MG hemithioacetal adduct. These data suggest that MG acts as a host cue that leads to GSH production and activation of PrfA.

Misinformation messages shared via WhatsApp in Mexico during the COVID-19 pandemic: an exploratory study.

Little is known about the role of WhatsApp in spreading misinformation during the start of the COVID-19 pandemic in Mexico. The aim of this study is to analyze the message content, format, authorship, time trends and social media distribution channels of misinformation in WhatsApp messages in Mexico. From March 18 to June 30, 2020 the authors collected all WhatsApp messages received via their personal contacts and their social networks that contained information about COVID-19. Descriptive and inferential statistics were used to analyze the scientifically inaccurate messages and the relationship between variables, respectively. Google image and video searches were carried out to identify sharing on other social media. Out of a total of 106 messages, the most frequently mentioned COVID-19 related message topics were prevention (20.0%), conspiracy (18.5%), therapy (15.4%) and origin of the virus (10.3%), changing throughout the pandemic according to users' concerns. Half of all WhatsApp messages were either images or videos. WhatsApp images were also shared on Facebook (80%) and YouTube (~50%). Our findings indicate that the design of information and health promotion campaigns requires to be proactive in adapting to the changes in message content and format of misinformation shared through encrypted social media.

As an encrypted social media platform with hardly accessible content, little is known about the role of WhatsApp in spreading misinformation messages (either false or misleading information) during the COVID-19 pandemic in Mexico. In this study, researchers studied the content, format, time and channel of distribution of WhatsApp messages containing information about COVID-19 collected via their personal contacts and their social networks from March 18 to June 30, 2020. Half of all messages were visually-appealing and the content changed according to the population´s concerns. WhatsApp messages were also distributed in several other social media platforms. Understanding the format and content of misinformation may help to design dynamic health information and promotion campaigns against it. Regulations of public social media such as Youtube can have a positive impact on WhatsApp.

Pharmacokinetic, sedative, and physiological effects of oral compounded formulations of trazodone alone or in combination with gabapentin in male cats.

Trazodone and gabapentin are common oral sedatives in cats, used alone or combined, but no pharmacokinetic studies exist for trazodone in this species. The objective of this study was to determine the pharmacokinetics of oral trazodone (T) alone, or in combination with gabapentin (G) in healthy cats. Cats (n = 6) were randomly allocated to receive T (3 mg/kg) intravenously (IV), T (5 mg/kg) orally (PO), or T (5 mg/kg) and G (10 mg/kg) PO with a 1-week washout period between treatments. Heart rate, respiratory rate, indirect blood pressure, and level of sedation were assessed, and venous blood samples were collected serially over 24 h. Analysis of plasma trazodone concentration was performed using liquid chromatography-tandem mass spectrometry (LC-MS/MS). Oral T administration resulted in a bioavailability of 54.9(7-96)%, and 17.2(11-25)% when administered with G. Tmax 0.17 (0.17-0.5) and 0.17 (0.17-0.75) h; Cmax 1.67 ± 0.91 and 1.22 ± 0.54 µg/mL, AUC 5.23 (2.0-18.76) and 2.37 (1.17-7.80) h*µg/mL; T1/2 5.12 ± 2.56 and 4.71 ± 1.07 h; for T and TG, respectively. Sedation was significant when compared to baseline in all groups from 20 or 45 min to 8 h indicating some lag between peak plasma concentration and sedative effects. Physiological variables remained within normal limits. This study concludes that oral trazodone is rapidly absorbed in healthy cats. Addition of gabapentin did not result in more profound sedation, showing no clinical advantage of combining these drugs in this study population.

Near-infrared fluorescence cholangiography in dogs: A pilot study.

OBJECTIVE: To determine the effect of indocyanine green (ICG) dose and timing of administration on near-infrared fluorescence (NIRF) imaging of the normal canine biliary tree. STUDY DESIGN: Preclinical prospective study. ANIMALS: Eight purpose-bred beagles. METHODS: The dogs were randomized to receive two of four intravenous ICG dose (low [L]:0.05 mg/kg or high [H]:0.25 mg/kg)/time (0 and 3 h prior to NIRF) combinations. NIRF images were collected every 10 min for 120 min. Target (cystic duct)-to-background (liver) ratios were calculated for all timepoints and compared. RESULTS: ICG cholangiography was successful in all dogs. The contrast ratio was above 1 in the L0 group by 20 min and reached its peak at 100 min. In the H0 group, the ratio was above 1 by 60 min and reached its peak at 90 min. Contrast ratios above 2 (fluorescence twice as bright in the cystic duct compared to the liver) were maintained from 180 to 300 min for L3 and H3 and was achieved after 80 min for L0. CONCLUSION: Low dose ICG provided better ratios early after injection compared to the high dose which remained highly concentrated in the liver tissue after injection. Both doses provided excellent visualization of the biliary tree at 3 h post injection, low dose ICG provided better ratios from 3 to 5 h post injection. Based on these results, 0.05 mg/kg of ICG administered at anesthetic premedication, or as early as 3 h prior to laparoscopic surgery should yield optimal fluorescence images. CLINICAL SIGNIFICANCE: This study provides guidelines for NIRF cholangiography in clinically normal dogs.

Some cardiopulmonary effects of capnoperitoneum in anesthetized guinea pigs (Cavia porcellus): spontaneous ventilation versus intubation and mechanical ventilation.

OBJECTIVE: To compare cardiopulmonary variables and blood gas analytes in guinea pigs (Cavia porcellus) during anesthesia with and without abdominal carbon dioxide (CO2) insufflation at intra-abdominal pressures (IAPs) 4 and 6 mmHg, with and without endotracheal intubation. STUDY DESIGN: Prospective experimental trial. ANIMALS: A total of six intact female Hartley guinea pigs. METHODS: A crossover study with sequence randomization for IAP and intubation status was used. The animals were sedated with intramuscular midazolam (1.5 mg kg-1) and buprenorphine (0.2 mg kg-1) and anesthetized with isoflurane, and an abdominal catheter was inserted for CO2 insufflation. Animals with endotracheal intubation were mechanically ventilated and animals maintained using a facemask breathed spontaneously. After 15 minutes of insufflation, the following variables were obtained at each IAP: pulse rate, respiratory rate, rectal temperature, oxygen saturation, end-tidal CO2 (intubated only), peak inspiratory pressure (intubated only), noninvasive blood pressure and blood gas and electrolyte values, with a rest period of 5 minutes between consecutive IAPs. After 4 weeks, the procedure was repeated with the guinea pigs assigned the opposite intubation status. RESULTS: Intubated guinea pigs had significantly higher pH and lower partial pressure of CO2 in cranial vena cava blood (PvCO2) than nonintubated guinea pigs. An IAP of 6 mmHg resulted in a significantly higher PvCO2 (65.9 ± 19.0 mmHg; 8.8 ± 2.5 kPa) than at 0 (53.2 ± 17.2 mmHg; 7.1 ± 2.3 kPa) and 4 mmHg (52.6 ± 10.8 mmHg; 7.01 ± 1.4 kPa), mean ± standard deviation, with intubated and nonintubated animals combined. CONCLUSIONS AND CLINICAL RELEVANCE: Although the oral anatomy of guinea pigs makes endotracheal intubation difficult, capnoperitoneum during anesthesia induces marked hypercapnia in the absence of mechanical ventilation. An IAP of 4 mmHg should be further evaluated for laparoscopic procedures in guinea pigs because hypercapnia may be less severe than with 6 mmHg.

Retrospective evaluation of acute hyperkalemia of unknown origin during general anesthesia in dogs.

OBJECTIVE: To report and characterize cases of acute hyperkalemia of unknown origin in dogs under anesthesia. STUDY DESIGN: Multicentric retrospective clinical study. ANIMALS: Medical records of 19 client-owned dogs that developed acute hyperkalemia during anesthesia. METHODS: Anesthetic records of dogs developing acute hyperkalemia from January 2015 to December 2022 were evaluated. Data collected included demographics, duration of anesthesia until the episode, electrolytes and blood gas measurements, electrocardiogram (ECG) abnormalities, drugs used as part of the anesthetic protocol, hyperkalemia treatment and outcome. RESULTS: A total of 13 cases met the inclusion criteria with documented acute hyperkalemia with no apparent underlying cause during anesthesia. Dogs were [mean ± standard deviation (range)] 6.5 ± 5.0 (3-10) years old and weighed 18.0 ± 14.3 (5.1-40.0) kg. All dogs were administered dexmedetomidine and an opioid as part of the premedication. All dogs had inhalation anesthesia of >60 minutes' duration. The first clinical sign was bradycardia that was minimally responsive to anticholinergic administration and was often accompanied by moderate/severe hypotension. These signs were rapidly followed by ECG changes compatible with hyperkalemia and/or cardiac arrest. Rapid identification and treatment for hyperkalemia, with or without dexmedetomidine reversal, resulted in survival of 12 dogs and one fatality. CONCLUSIONS AND CLINICAL RELEVANCE: Unknown origin hyperkalemia is a life-threatening complication that can occur during general anesthesia. In healthy dogs, preanesthetic administration of dexmedetomidine in association with an opioid and followed by inhalation anesthesia of more than 1 hour duration may predispose to this complication. A sudden decrease in heart rate >90 minutes after dexmedetomidine administration, or ECG changes, may warrant measurement of blood potassium concentrations.

Phytostabilization of trace elements and 13C isotope composition of Atriplex atacamensis Phil. cultivated in mine tailings treated with organic amendments.

Mining generates large quantities of mineral processing wastes that are typically stored in mine tailings (MT) ponds. Long-term exposure of the surrounding areas to the material from the tailings ponds has been reported to have adverse effects on both human health and the environment. The purpose of this study was to evaluate the ability of Atriplex atacamensis Phil. to phytostabilize metals (Cu, Fe, Mn, and Zn) and sulfur (S) when grown directly on mine tailings with and without compost (C) and humic substance (HS). The stress status of A. atacamensis Phil. was also evaluated through the 13C isotopic composition of bulk leaves. A 120-day greenhouse experiment was conducted and three treatments were evaluated: (i) MT without any amendments (control), (ii) MT + C (dose: 89 ton ha-1), and (iii) MT + HS (0.72 ton ha-1). Mine tailings material exhibited low salinity, alkaline pH, high extractable S-SO4 concentrations, and low fertility; total Fe, Mn, and Zn concentrations were within the reference range for mine tailings, but total Cu concentrations were high at 1860 ± 236 mg kg-1. The HS had higher pH, EC, CEC, and available concentrations of N, P, and K than compost, while S-SO4 concentrations were similar in both amendments. 13C NMR analysis showed that the HS contained more alkyl, aromatic, and phenolic groups, while the compost was dominated by O-alkyl and carboxyl groups. At the end of the experiment, the MT + C treatment achieved a significant decrease in Cu, Fe, and Mn concentrations in the roots and aboveground parts of A. atacamensis Phil. and an increase in Zn values in both tissues. Both amendments increased the sulfur content in the aboveground parts, while metal concentrations under the HS treatment proved similar to control. Furthermore, the δ13CV-PDB values obtained in this study indicate that the organic amendments did not cause additional physiological stress to the plants compared to the MT treatment. Overall, A. atacamensis Phil. was shown to have the ability to phytostabilize metals and sulfur, making it a potential candidate species for in situ evaluation of the phytostabilization process on mine tailings.

para-Aminobenzoic Acid Biosynthesis Is Required for Listeria monocytogenes Growth and Pathogenesis.

Biosyntheses of para-aminobenzoic acid (PABA) and its downstream folic acid metabolites are essential for one-carbon metabolism in all life forms and the targets of sulfonamide and trimethoprim antibiotics. In this study, we identified and characterized two genes (pabA and pabBC) required for PABA biosynthesis in Listeria monocytogenes. Mutants in PABA biosynthesis were able to grow normally in rich media but not in defined media lacking PABA, but growth was restored by the addition of PABA or its downstream metabolites. PABA biosynthesis mutants were attenuated for intracellular growth in bone marrow-derived macrophages, produced extremely small plaques in fibroblast monolayers, and were highly attenuated for virulence in mice. PABA biosynthesis genes were upregulated upon infection and induced during growth in broth in a strain in which the master virulence regulator, PrfA, was genetically locked in its active state (PrfA*). To gain further insight into why PABA mutants were so attenuated, we screened for transposon-induced suppressor mutations that formed larger plaques. Suppressor mutants in relA, which are predicted to have higher levels of (p)ppGpp, and mutants in codY, which is a GTP-binding repressor of many biosynthetic genes, partially rescued the plaque defect but, notably, restored the capacity of the mutants to escape from phagosomes and induce the polymerization of host cell actin. However, these suppressor mutant strains remained attenuated for virulence in mice. These data suggest that even though folic acid metabolites exist in host cells and might be available during infection, de novo synthesis of PABA is required for L. monocytogenes pathogenesis.

Comparative genomics and proteomics of Eubacterium maltosivorans: functional identification of trimethylamine methyltransferases and bacterial microcompartments in a human intestinal bacterium with a versatile lifestyle.

Eubacterium maltosivorans YIT is a human intestinal isolate capable of acetogenic, propionogenic and butyrogenic growth. Its 4.3-Mb genome sequence contains coding sequences for 4227 proteins, including 41 different methyltransferases. Comparative proteomics of strain YIT showed the Wood-Ljungdahl pathway proteins to be actively produced during homoacetogenic growth on H2 and CO2 while butyrogenic growth on a mixture of lactate and acetate significantly upregulated the production of proteins encoded by the recently identified lctABCDEF cluster and accessory proteins. Growth on H2 and CO2 unexpectedly induced the production of two related trimethylamine methyltransferases. Moreover, a set of 16 different trimethylamine methyltransferases together with proteins for bacterial microcompartments were produced during growth and deamination of the quaternary amines, betaine, carnitine and choline. Growth of strain YIT on 1,2-propanediol generated propionate with propanol and induced the formation of bacterial microcompartments that were also prominently visible in betaine-grown cells. The present study demonstrates that E. maltosivorans is highly versatile in converting low-energy fermentation end-products in the human gut into butyrate and propionate whilst being capable of preventing the formation of the undesired trimethylamine by converting betaine and other quaternary amines in bacterial microcompartments into acetate and butyrate.

Analysis of physical learning spaces in a university hospital: A case study.

BACKGROUND: Attributes of physical learning spaces can facilitate or hinder learning. There are few studies about this topic in hospitals. The objective of this study was to explore the characteristics of physical learning spaces in a university hospital. METHODS: The setting was a large research-oriented public university hospital in Mexico City, affiliated with the National Autonomous University of Mexico. An intrinsic case study design was conducted with two instruments: a questionnaire to identify physical learning spaces and their attributes; the Learning Space Rating System (LSRS), an instrument used to evaluate spaces' characteristics that promote effective learning. RESULTS: 49 medical students and 60 internal medicine residents responded to the questionnaire. The attributes with the highest importance for students were: instructor physical availability, silence, comfort of the seating furniture, and Internet access. The study authors directly performed an evaluation of the learning spaces using the LSRS tool, the sections' scores were: clinical ward discussion room (74%), external consultation unit (71%), auditorium (70%), classroom (68%), library (66%), and the hospitalization room (61%). CONCLUSIONS: Physical learning spaces in medical training can be formally assessed to identify the attributes that students consider relevant for learning and provide needed information for redesign and reuse of spaces. Medical education scholars and trainees should be involved in the design and evaluation of university and hospital buildings.

A Review on Current Strategies for Extraction and Purification of Hyaluronic Acid.

Since it is known that hyaluronic acid contributes to soft tissue growth, elasticity, and scar reduction, different strategies of producing HA have been explored in order to satisfy the current demand of HA in pharmaceutical products and formulations. The current interest deals with production via bacterial and yeast fermentation and extraction from animal sources; however, the main challenge is the right extraction technique and strategy since the original sources (e.g., fermentation broth) represent a complex system containing a number of components and solutes, which complicates the achievement of high extraction rates and purity. This review sheds light on the main pathways for the production of HA, advantages, and disadvantages, along with the current efforts in extracting and purifying this high-added-value molecule from different sources. Particular emphasis has been placed on specific case studies attempting production and successful recovery. For such works, full details are given together with their relevant outcomes.

Description and validation of a new descriptive and multiparametric numeric rating scale to assess sedation in cats.

The objective of this study was to design and assess the validity and reliability of a new feline multiparametric sedation scale (FMSS). A total of 89 household cats were recruited, enabling a total of 534 sedation assessments. Every assessment was performed by 3 blinded observers with varying expertise levels (Level 1: Student; Level 2: RVT; Level 3: ACVAA diplomate or senior resident). For comparison purposes, a visual analogue scale (VAS) and a Simple Qualitative Scale (SQS) were also used concurrently, with the VAS considered the gold standard. The new scale had excellent inter-observer agreement among experience groups with weighted Kappa scores of 0.84 (Levels 1 versus 2), 0.82 (Levels 2 versus 3), and 0.84 (Levels 1 versus 3), with P < 0.0001 for all comparisons. There was a high degree of association between FMSS and VAS (r = 0.90, P < 0.0001) and between FMSS and SQS (r = 0.89, P < 0.0001). Final FMSS numerical values were paired with levels of sedation with None = 0 (0 to 5), Mild = 4 (1 to 7), Moderate = 6 (2 to 10), and Profound = 12 (7 to 12); furthermore, differences were detected between pre- and post-sedation evaluations (P = 0.001). This scale demonstrated internal consistency and sensitivity even when evaluating drugs or doses with minimal sedative effects and there was very strong interrater reliability, independent of experience level. Based on this clinical study, we concluded that the use of this sedation scale is appropriate when objective numerical sedation quantification is required, in either a clinical or research setting.

Description et validation d'une nouvelle échelle d'évaluation numérique descriptive et multiparamétrique pour évaluer la sédation chez le chat. L'objectif de cette étude était de concevoir et d'évaluer la validité et la fiabilité d'une nouvelle échelle de sédation multiparamétrique féline (FMSS). Un total de 89 chats domestiques a été recruté, permettant un total de 534 évaluations de sédation. Chaque évaluation a été effectuée par trois observateurs en aveugle avec différents niveaux d'expertise (Niveau 1 : étudiant; Niveau 2 : RVT; Niveau 3 : diplomate de l'ACVAA ou résident senior). À des fins de comparaison, une échelle visuelle analogique (VAS) et une échelle qualitative simple (SQS) ont également été utilisées simultanément, VAS étant considérée comme l'étalon. La nouvelle échelle présentait un excellent accord inter-observateurs parmi les groupes d'expérience avec des scores Kappa pondérés de 0,84 (niveaux 1 versus 2), 0,82 (niveaux 2 versus 3) et 0,84 (niveaux 1 versus 3), avec P < 0,0001 pour toutes les comparaisons. Il y avait un degré élevé d'association entre FMSS et VAS (r = 0,90, P < 0,0001) et entre FMSS et SQS (r = 0,89, P < 0,0001). Les valeurs numériques FMSS finales ont été appariées avec les niveaux de sédation avec Aucun = 0 (0 à 5), Léger = 4 (1 à 7), Modéré = 6 (2 à 10) et Profond = 12 (7 à 12); en outre, des différences ont été détectées entre les évaluations pré- et post-sédation (P = 0,001). Cette échelle a démontré une cohérence interne et une sensibilité même lors de l'évaluation de médicaments ou de doses avec des effets sédatifs minimes et il y avait une très forte fiabilité inter-évaluateur, indépendamment du niveau d'expérience. Sur la base de cette étude clinique, nous avons conclu que l'utilisation de cette échelle de sédation est appropriée lorsqu'une quantification numérique objective de la sédation est requise, dans un cadre clinique ou de recherche.(Traduit par Dr Serge Messier).