A Neutrophil Timer Coordinates Immune Defense and Vascular Protection.

Neutrophils eliminate pathogens efficiently but can inflict severe damage to the host if they over-activate within blood vessels. It is unclear how immunity solves the dilemma of mounting an efficient anti-microbial defense while preserving vascular health. Here, we identify a neutrophil-intrinsic program that enabled both. The gene Bmal1 regulated expression of the chemokine CXCL2 to induce chemokine receptor CXCR2-dependent diurnal changes in the transcriptional and migratory properties of circulating neutrophils. These diurnal alterations, referred to as neutrophil aging, were antagonized by CXCR4 (C-X-C chemokine receptor type 4) and regulated the outer topology of neutrophils to favor homeostatic egress from blood vessels at night, resulting in boosted anti-microbial activity in tissues. Mice engineered for constitutive neutrophil aging became resistant to infection, but the persistence of intravascular aged neutrophils predisposed them to thrombo-inflammation and death. Thus, diurnal compartmentalization of neutrophils, driven by an internal timer, coordinates immune defense and vascular protection.

Vitamin D and monoclonal gammopathy of undetermined significance (MGUS) among U.S. Black women.

PURPOSE: Risk factors for monoclonal gammopathy of undetermined significance (MGUS), the asymptomatic precursor to multiple myeloma, are largely unknown. We hypothesized that low vitamin D levels might be associated with higher MGUS prevalence in a national cohort of U.S. Black women. METHODS: We screened archived serum samples (collected 2014-2017) from 3896 randomly selected participants in the Black Women's Health Study ages 50-79 for evidence of MGUS; samples had been assayed for 25-hydroxyvitamin D [25(OH)D] shortly after blood draw. We used logistic regression to estimate odds ratios (OR) and 95% confidence intervals (CI) for the association between 25(OH)D level and MGUS status, adjusting for age, body mass index, and season of blood draw. RESULTS: We identified 334 MGUS cases (8.6%) in the study population. The adjusted OR comparing women with vitamin D deficiency (25(OH)D < 20 ng/mL) to those with 25(OH)D levels ≥ 30 ng/mL was 1.27 (95% CI: 0.95, 1.72). CONCLUSION: MGUS was more prevalent among Black women with vitamin D deficiency compared to those with 25(OH)D ≥ 30 ng/mL; however, the association was not statistically significant. Future prospective studies are warranted to clarify the possible association between vitamin D and MGUS.

Functional and Structural Analysis Reveals Distinct Biological Roles of Plant Synaptotagmins in Response to Environmental Stress.

Endoplasmic reticulum-plasma membrane contact sites (ER-PM CSs) are evolutionarily conserved membrane domains found in all eukaryotes, where the ER closely interfaces with the PM. This short distance is achieved in plants through the action of tether proteins such as synaptotagmins (SYTs). Arabidopsis comprises five SYT members (SYT1-SYT5), but whether they possess overlapping or distinct biological functions remains elusive. SYT1, the best-characterized member, plays an essential role in the resistance to abiotic stress. This study reveals that the functionally redundant SYT1 and SYT3 genes, but not SYT5, are involved in salt and cold stress resistance. We also show that, unlike SYT5, SYT1 and SYT3 are not required for Pseudomonas syringae resistance. Since SYT1 and SYT5 interact in vivo via their SMP domains, the distinct functions of these proteins cannot be caused by differences in their localization. Interestingly, structural phylogenetic analysis indicates that the SYT1 and SYT5 clades emerged early in the evolution of land plants. We also show that the SYT1 and SYT5 clades exhibit different structural features in their SMP and Ca2+ binding of their C2 domains, rationalizing their distinct biological roles.

A 14-year review (2007-2020) of helminthiasis epidemiology in a hospital in Southern Madrid, Spain.

PURPOSE: Vast majority of helminth diseases remain neglected tropical diseases (NTDs), causing significant morbidity. The widespread and periodic distribution of antiparasitic drugs, remains the cornerstone for controlling these diseases. In Spain, most helminthiasis cases are imported, and suspicion and diagnosis have become increasingly important. Our primary objective is to present the epidemiological landscape of helminthiasis diagnoses within our facility, while also detailing the demographic characteristics of the affected population. METHODS: A retrospective study was conducted at the Hospital Universitario Severo Ochoa (HUSO) from January 1, 2007, to December 31, 2020, encompassing all diagnosed cases of helminthiasis during this period. Comprehensive epidemiological, clinical, and microbiological data were gathered for all diagnosed patients. The study population comprised patients receiving treatment at the HUSO, as well as those receiving treatment at the Leganés and Fuenlabrada Primary Care Units. Subsequently, descriptive and comparative statistics were performed, comparing Spanish and foreign patients. RESULTS: During this period, a total of 952 patients were diagnosed with some form of helminthiasis. Among them, 495 were Spanish, and 457 were foreign. The total number of helminths identified, including patients with multiple infections, was 1,010. Significant differences were observed between Africans and Americans in terms of age distribution, with a higher prevalence among Africans in the 0-15 age range and among Americans in the 31-60 age range. Variations were noted in the distribution of helminths, with S. stercoralis significantly affecting Americans. For Spanish patients, the presence of Trichuris trichiura and S. stercoralis was significantly associated with eosinophilia, whereas among foreign patients, it was associated with Trichuris trichiura, Ascaris lumbricoides among others. Regarding symptoms, skin manifestations were more frequent among Spanish, while digestive were more common among foreigners. CONCLUSIONS: This study offers crucial epidemiological insights into helminth infections observed over time in a Madrid hospital. Although the prevalence of helminth infections has been decreasing, there is still a need for screening and diagnosing foreign patients.

Imported malaria in a non-endemic country: sixteen years of cases in a hospital in the South of Madrid, Spain.

PURPOSE: Malaria keeps on being a serious global health threat, especially in many tropical countries, where it is endemic. Also in non-endemic countries, like Spain, malaria is an issue that requires attention due to the presence of imported cases. METHODS: This is a retrospective study, including all patients diagnosed with malaria at Severo Ochoa University Hospital from 2006 to 2022, being classified according to: (I) their type of stay in an endemic area as visiting friends and relatives (VFR), migrants of recent arrival (MRA), or tourism and business (T&B), and (II) the mode of presentation as microscopic (MM) or submicroscopic (SMM) malaria. RESULTS: In this study, 132 patients (23.7% of all suspected) were diagnosed with malaria. The PCR was the most sensitive technique (99.2%), followed by antigen detection (78.8%) and microscopy (75%), with Plasmodium falciparum being the predominant species (94.7%). VFR was the largest group infected with malaria (69.7%), mostly symptomatic (98.2%) and presenting MM (90.2%). Instead, MRA patients (25%) presented milder (47.4%) or no symptoms (31.6%) and higher cases of SMM (42.4%). Coinfection with another imported pathogen was present in 19 patients (14.4%), being MRA more frequently coinfected (30.3%) CONCLUSION: This study shows the need for establishing systems for VFRs to attend pre-travel consultations to reduce malaria imported risk. In the case of MRA, screening for imported diseases should be conducted upon their arrival. Finally, we highlight two cases of co-infection with imported viruses, showing that presence of symptoms resembling malaria from another imported pathogen does not exclude malaria.

Microwave-Assisted Synthesis of CdS-MOF MIL-101 (Fe) Composite: Characterization and Photocatalytic Performance.

The present study outlines the synthesis of cadmium sulfide-metal-organic framework (CdS-MOF) MIL-101 (Fe) heterojunctions achieved via fast microwave-assisted reactions. Thus, different CdS-MOF MIL-101 (Fe) ratios were prepared to study their effectiveness as photocatalysts. These compounds were employed in the photocatalytic degradation of methylene blue (MB) under UV and visible irradiation. Structural, morphological, textural, compositional, and optical properties of the synthesized compounds determined by X-ray diffraction (XRD), scanning electron microscopy (SEM), X-ray photoelectron spectroscopy (XPS), UV-vis spectroscopy, Raman spectroscopy, and Fourier transform infrared (FTIR) spectroscopy were utilized to characterize the structural, morphological, textural, compositional, and optical properties. Electrochemical impedance spectroscopy (EIS) was also employed to determine photovoltage and photocurrent densities. The resulting valence band offset (Vfb) and band gap energy values were utilized to construct an energy band scheme. Our research revealed that the CdS-MOF MIL-101 (Fe) heterojunction enhances the efficiency of electron-hole pair separation, thereby mitigating charge carrier recombination effects. Moreover, the type I electronic band structure established an efficient reaction mechanism, effectively suppressing the recombination of photogenerated electron-hole pairs. The photocatalytic system demonstrated exceptional behavior, achieving complete MB removal within 30 min of reaction time and exhibiting outstanding stability and reusability after four reaction cycles. These findings highlight the potential of the synthesized compounds in the field of wastewater treatment for organic pollutants, offering a promising alternative to current environmental issues.

High-Resolution Melting Assay to Detect the Mutations That Cause the Y132F and G458S Substitutions at the ERG11 Gene Involved in Azole Resistance in Candida parapsilosis.

BACKGROUND: Candida parapsilosis is a pathogenic yeast that has reduced susceptibility to echinocandins and ranks as the second or third leading cause of candidaemia, depending on the geographical region. This yeast often causes nosocomial infections, which are frequently detected as outbreaks. In recent years, resistance to azoles in C. parapsilosis has increased globally, primarily due to the accumulation of mutations in the ERG11 gene. OBJECTIVES: In this study, we have developed an assay based on real-time PCR and high-resolution melting (HRM) curve analysis to detect two of the most prevalent mutations at ERG11 that confer resistance to fluconazole (Y132F and G458S). METHODS: We designed allele-specific oligonucleotides that selectively bind to either the wild type or mutated sequences and optimised the conditions to ensure amplification of the specific allele, followed by detection via high-resolution melting (HRM) analysis. RESULTS: The designed oligonucleotides to detect the Erg11Y132F and Erg11G458S mutations produced specific amplification of either WT or mutated alleles. We conducted a duplex real-time PCR combining oligonucleotides for the wild-type sequences in one mix, and oligonucleotides for the mutated alleles in another. Following this, we performed an analysis of the HRM curve to identify the amplified allele in each case. This technique was blindly evaluated on a set of 114 C. parapsilosis isolates, all of which were unequivocally identified using our approach. CONCLUSION: This technique offers a new method for the early detection of azole resistance mechanism in C. parapsilosis.

Protein-Protein Binding Kinetics by Biolayer Interferometry.

The specific kinetics and thermodynamics of protein-protein interactions underlie the molecular mechanisms of cellular functions; hence the characterization of these interaction parameters is central to the quantitative understanding of physiological and pathological processes. Many methods have been developed to study protein-protein interactions, which differ in various features including the interaction detection principle, the sensitivity, whether the method operates in vivo, in vitro, or in silico, the temperature control, the use of labels, immobilization, the amount of sample required, the number of measurements that can be accomplished simultaneously, or the cost. Bio-Layer Interferometry (BLI) is a label-free biophysical method to measure the kinetics of protein-protein interactions. Label-free interaction assays are a broad family of methods that do not require protein modifications (other than immobilization) or labels such as fusions with fluorescent proteins or transactivating domains or chemical modifications like biotinylation or reaction with radionuclides. Besides BLI, other label-free techniques that are widely used for determining protein-protein interactions include surface plasmon resonance (SPR), thermophoresis, and isothermal titration calorimetry (ITC), among others.

A case-control autopsy series of liver pathology associated with novel coronavirus disease (COVID-19).

BACKGROUND: Although severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) responsible for coronavirus disease 2019 (COVID-19) is most well-known for causing pulmonary injury, a significant proportion of patients experience hepatic dysfunction. The mechanism by which SARS-CoV2 causes liver injury is not fully understood. The goal of this study was to describe the hepatic pathology in a large cohort of deceased patients with COVID-19 as compared to a control group of deceased patients without COVID-19. METHODS: Consented autopsy cases at two institutions were searched for documentation of COVID-19 as a contributing cause of death. A group of consecutive consented autopsy cases during the same period, negative for SARS-CoV-2 infection, was used as a control group. The autopsy report and electronic medical records were reviewed for relevant clinicopathologic information. H&E-stained liver sections from both groups were examined for pertinent histologic features. Select cases underwent immunohistochemical staining for CD 68 and ACE2 and droplet digital polymerase chain reaction (ddPCR) assay for evaluation of SARS-CoV2 RNA. RESULTS: 48 COVID-19 positive patients (median age 73, M:F 3:1) and 40 COVID-19 negative control patients (median age 67.5, M:F 1.4:1) were included in the study. The COVID-19 positive group was significantly older and had a lower rate of alcoholism and malignancy, but there was no difference in other comorbidities. The COVID-19 positive group was more likely to have received steroids (75.6 % vs. 36.1 %, p < 0.001). Hepatic vascular changes were seen in a minority (10.6 %) of COVID-19 positive cases. When all patients were included, there were no significant histopathologic differences between groups, but when patients with chronic alcoholism were excluded, the COVID-19 positive group was significantly more likely to have steatosis (80.9 % vs. 50.0 %, p = 0.004) and lobular inflammation (45.7 % vs. 20.7 %, p = 0.03). Testing for viral RNA by ddPCR identified 2 of the 18 (11.1 %) COVID-19 positive cases to have SARS-CoV-2 RNA detected within the liver FFPE tissue. CONCLUSIONS: The most significant findings in the liver of COVID-19 positive patients were mild lobular inflammation and steatosis. The high rate of steroid therapy in this population may be a possible source of steatosis. Hepatic vascular alterations were only identified in a minority of patients and did not appear to play a predominant role in COVID-19 mediated hepatic injury. Low incidence of SARS-CoV-2 RNA positivity in liver tissue in our cohort suggests hepatic injury in the setting of COVID-19 may be secondary in nature.

Determination of change of direction deficit thresholds across a spectrum of angles in basketball players.

Change of direction deficit (CODD) offers valuable insights into a player's balance between linear and multidirectional speed. However, there are still no established reference values for CODD. The objectives of this study were to determine CODD thresholds for various change of direction angles in basketball players according to gender and analyse the relationships between CODD and execution time in speed tests. One hundred and thirty basketball players (46% female; age: 23.7 ± 5.29 years; height: 189.1 ± 11.1 cm; body mass: 84.3 ± 15.7 kg) undertook 10-m linear and change of direction speed test at 45º, 90º and 180º. A k-means cluster analysis was conducted to standardise CODD thresholds and a one-way analysis of variance to identify the differences between clusters. The results revealed angulation-specific CODD thresholds, ranging from 3% to 8%, 17% to 25% and 43% to 51% for 45º, 90º and 180º cutting angles, respectively for the pooled sample. Furthermore, differences inter-clusters (p < 0.05) were observed for execution time at all cutting angles for both genders. Therefore, strength and conditioning coaches are encouraged to assess CODD as a highly valid variable for evaluating change of direction performance and to use current CODD thresholds to tailor training programmes according to each athlete's needs.

Breast density quantification in dual-energy mammography using virtual anthropomorphic phantoms.

PURPOSE: Breast density is a significant risk factor for breast cancer and can impact the sensitivity of screening mammography. Area-based breast density measurements may not provide an accurate representation of the tissue distribution, therefore volumetric breast density (VBD) measurements are preferred. Dual-energy mammography enables volumetric measurements without additional assumptions about breast shape. In this work we evaluated the performance of a dual-energy decomposition technique for determining VBD by applying it to virtual anthropomorphic phantoms. METHODS: The dual-energy decomposition formalism was used to quantify VBD on simulated dual-energy images of anthropomorphic virtual phantoms with known tissue distributions. We simulated 150 phantoms with volumes ranging from 50 to 709 mL and VBD ranging from 15% to 60%. Using these results, we validated a correction for the presence of skin and assessed the method's intrinsic bias and variability. As a proof of concept, the method was applied to 14 sets of clinical dual-energy images, and the resulting breast densities were compared to magnetic resonance imaging (MRI) measurements. RESULTS: Virtual phantom VBD measurements exhibited a strong correlation (Pearson's r > 0.95 $r > 0.95$ ) with nominal values. The proposed skin correction eliminated the variability due to breast size and reduced the bias in VBD to a constant value of -2%. Disagreement between clinical VBD measurements using MRI and dual-energy mammography was under 10%, and the difference in the distributions was statistically non-significant. VBD measurements in both modalities had a moderate correlation (Spearman's ρ $\rho \ $ = 0.68). CONCLUSIONS: Our results in virtual phantoms indicate that the material decomposition method can produce accurate VBD measurements if the presence of a third material (skin) is considered. The results from our proof of concept showed agreement between MRI and dual-energy mammography VBD. Assessment of VBD using dual-energy images could provide complementary information in dual-energy mammography and tomosynthesis examinations.

A Video Mosaicing-Based Sensing Method for Chicken Behavior Recognition on Edge Computing Devices.

Chicken behavior recognition is crucial for a number of reasons, including promoting animal welfare, ensuring the early detection of health issues, optimizing farm management practices, and contributing to more sustainable and ethical poultry farming. In this paper, we introduce a technique for recognizing chicken behavior on edge computing devices based on video sensing mosaicing. Our method combines video sensing mosaicing with deep learning to accurately identify specific chicken behaviors from videos. It attains remarkable accuracy, achieving 79.61% with MobileNetV2 for chickens demonstrating three types of behavior. These findings underscore the efficacy and promise of our approach in chicken behavior recognition on edge computing devices, making it adaptable for diverse applications. The ongoing exploration and identification of various behavioral patterns will contribute to a more comprehensive understanding of chicken behavior, enhancing the scope and accuracy of behavior analysis within diverse contexts.

The Effect of Ultrasound on the Extraction and Functionality of Proteins from Duckweed (Lemna minor).

The inclusion of protein in the regular human diet is important for the prevention of several chronic diseases. In the search for novel alternative protein sources, plant-based proteins are widely explored from a sustainable and ecological point of view. Duckweed (Lemna minor), also known as water lentil, is an aquatic plant with potential applications for human consumption due to its protein content and carbohydrate contents. Among all the conventional and novel protein extraction methods, the utilization of ultrasound has attracted the attention of scientists because of its effects on improving protein extraction and its functionalities. In this work, a Box-Behnken experimental design was proposed to optimize the alkaline extraction of protein from duckweed. In addition, an exploration of the effects of ultrasound on the morphological, structural, and functional properties of the extracted protein was also addressed. The optimal extraction parameters were a pH of 11.5 and an ultrasound amplitude and processing time of 60% and 20 min, respectively. These process conditions doubled the protein content extracted in comparison to the value from the initial duckweed sample. Furthermore, the application of ultrasound during the extraction of protein generated changes in the FTIR spectra, color, and structure of the duckweed protein, which resulted in improvements in its solubility, emulsifying properties, and foaming capacity.

Effectiveness of Mindfulness Therapy as a Complement to Pharmacological Treatment in the Control of Blood Pressure in Hypertensive Adults: A Quasi-Experimental Study in Lima, Peru.

OBJECTIVE: To evaluate the effectiveness of mindfulness therapy as an adjunct to pharmacological treatment for improving blood pressure (BP) control in hypertensive adult patients at a health center in Lima, Peru. METHOD: This study employed a quasi-experimental design with a quantitative and longitudinal cohort approach. The participants included 50 adults who were diagnosed with hypertension and who regularly attended the health center in the Villa El Salvador district of Lima, Peru. The data were collected using three instruments: the Depression Anxiety Stress Scale (DASS 21), validated in Peru; a sociodemographic questionnaire; and a BP recording sheet. The Wilcoxon test was utilized to assess the effectiveness of mindfulness therapy on BP control, while the nonparametric Friedman test was used to evaluate the effectiveness of mindfulness therapy considering confounding factors. RESULTS: Therapy resulted in favorable outcomes for BP control. After eight sessions, the average systolic BP decreased from 122.8 to 115.0 mm Hg (a reduction of 7%), and the average diastolic BP decreased from 78.1 to 69.9 mm Hg (a reduction of 11%). CONCLUSIONS: Mindfulness is an effective complementary therapy for controlling BP in hypertensive adults. It also helps reduce confounding factors such as stress, anxiety, and depression.

Broad Protection against Invasive Fungal Disease from a Nanobody Targeting the Active Site of Fungal ß-1,3-Glucanosyltransferases.

Invasive fungal disease accounts for about 3.8 million deaths annually, an unacceptable rate that urgently prompts the discovery of new knowledge-driven treatments. We report the use of camelid single-domain nanobodies (Nbs) against fungal ß-1,3-glucanosyltransferases (Gel) involved in ß-1,3-glucan transglycosylation. Crystal structures of two Nbs with Gel4 from Aspergillus fumigatus revealed binding to a dissimilar CBM43 domain and a highly conserved catalytic domain across fungal species, respectively. Anti-Gel4 active site Nb3 showed significant antifungal efficacy in vitro and in vivo prophylactically and therapeutically against different A. fumigatus and Cryptococcus neoformans isolates, reducing the fungal burden and disease severity, thus significantly improving immunocompromised animal survival. Notably, C. deneoformans (serotype D) strains were more susceptible to Nb3 and genetic Gel deletion than C. neoformans (serotype A) strains, indicating a key role for ß-1,3-glucan remodelling in C. deneoformans survival. These findings add new insight about the role of ß-1,3-glucan in fungal biology and demonstrate the potential of nanobodies in targeting fungal enzymes to combat invasive fungal diseases.

Progressive multifocal leukoencephalopathy: a retrospective study of the last 12 years in a tertiary-care hospital.

Our study aims to report on the demographic, incidence rate (IR), clinical, and microbiological characteristics of PML patients diagnosed in our tertiary-care hospital over the past 12 years. In this retrospective observational study, we reviewed all requests for JCPyV PCR in CSF from patients with suspected PML. We collected demographic, clinical, and microbiological data of patients diagnosed with PML. Since 2018, real-time quantitative PCR has been used, whereas prior to 2018, samples were sent to our National Reference Center for qualitative diagnosis. Thirteen patients were diagnosed with PML, with 10 of them having a definitive diagnosis and 3 classified as a possible diagnosis with negative PCR results. Eleven patients had advanced HIV, one had non-Hodgkin's lymphoma, and one had systemic lupus erythematosus. Most of the white matter lesions were located at the cerebral level, although the parenchyma and cerebellum were also affected. The most frequent symptoms were behavioral disorders and hemiparesis. The viral load of JCPyV in cerebrospinal fluid was < 1000 copies/mL in three patients. Six patients received compassionate treatment, and all six patients with definitive PML diagnosis died. Although advanced HIV patients were the most affected by PML in our study, it should also be considered in patients with other underlying diseases. While current PCR tests offer high sensitivity and specificity, false negatives can occur. The prognosis of the disease remains poor, and early multidisciplinary diagnosis-including clinical, microbiological, and neuroimaging assessments-remains crucial for improving neurological damage and prognosis.

Subgingival host-microbiome metatranscriptomic changes following scaling and root planing in grade II/III periodontitis.

AIM: To assess the effects of scaling and root planing (SRP) on the dynamics of gene expression by the host and the microbiome in subgingival plaque samples. MATERIALS AND METHODS: Fourteen periodontitis patients were closely monitored in the absence of periodontal treatment for 12 months. During this period, comprehensive periodontal examination and subgingival biofilm sample collection were performed bi-monthly. After 12 months, clinical attachment level (CAL) data were compiled and analysed using linear mixed models (LMM) fitted to longitudinal CAL measurements for each tooth site. LMM classified the sites as stable (S), progressing (P), or fluctuating (F). After the 12-month visit, subjects received SRP, and at 15 months they received comprehensive examination and supportive periodontal therapy. Those procedures were repeated at the 18-month visit, when patients were also sampled. Each patient contributed with one S, one P, and one F site collected at the 12- and 18-month visits. Samples were analysed using Dual RNA-Sequencing to capture host and bacterial transcriptomes simultaneously. RESULTS: Microbiome and host response behaviour were specific to the site's progression classification (i.e., S, P, or F). Microbial profiles of pre- and post-treatment samples exhibited specific microbiome changes, with progressing sites showing the most significant changes. Among them, Porphyromonas gingivalis was reduced after treatment, while Fusobacterium nucleatum showed an increase in proportion. Transcriptome analysis of the host response showed that interleukin (IL)-17, TNF signalling pathways, and neutrophil extracellular trap formation were the primary immune response activities impacted by periodontal treatment. CONCLUSIONS: SRP resulted in a significant "rewiring" of host and microbial activities in the progressing sites, while restructuring of the microbiome was minor in stable and fluctuating sites.

Neurofilament light chain in spinal fluid and plasma in multiple system atrophy: a prospective, longitudinal biomarker study.

PURPOSE: There is a critical need for reliable diagnostic biomarkers as well as surrogate markers of disease progression in multiple system atrophy (MSA). Neurofilament light chain (NfL) has been reported to potentially meet those needs. We therefore sought to explore the value of NfL in plasma (NfL-p) in contrast to cerebrospinal fluid (NfL-c) as a diagnostic marker of MSA, and to assess NfL-p and NfL-c as markers of clinical disease progression. METHODS: Well-characterized patients with early MSA (n = 32), Parkinson's disease (PD; n = 21), and matched controls (CON; n = 15) were enrolled in a prospective, longitudinal study of synucleinopathies with serial annual evaluations. NfL was measured using a high-sensitivity immunoassay, and findings were assessed by disease category and relationship with clinical measures of disease progression. RESULTS: Measurements of NfL-c were highly reproducible across immunoassay platforms (Pearson, r = 0.99), while correlation between NfL-c and -p was only moderate (r = 0.66). NfL was significantly higher in MSA compared with CON and PD; the separation was essentially perfect for NfL-c, but there was overlap, particularly with PD, for NfL-p. While clinical measures of disease severity progressively increased over time, NfL-c and -p remained at stable elevated levels within subjects across serial measurements. Neither change in NfL nor baseline NfL were significantly associated with changes in clinical markers of disease severity. CONCLUSIONS: These findings confirm NfL-c as a faithful diagnostic marker of MSA, while NfL-p showed less robust diagnostic value. The significant NfL elevation in MSA was found to be remarkably stable over time and was not predictive of clinical disease progression.

Metronidazole for Prevention of Pelvic Cellulitis and Abscess after Laparoscopic Hysterectomy: A Triple-blinded, Randomized, Placebo-controlled Clinical Trial.

STUDY OBJECTIVE: To determine whether a postoperative 5-day treatment schedule with vaginal metronidazole added to conventional antibiotic prophylaxis with 2 g cefazolin modifies the risk of pelvic cellulitis (PC) and pelvic abscess (PA) after total laparoscopic hysterectomy (TLH). DESIGN: A randomized, controlled, triple-blind, multicenter clinical trial. SETTING: Two centers dedicated to minimally invasive gynecologic surgery in Colombia. PATIENTS: A total of 574 patients were taken to TLH because of benign diseases. INTERVENTION: Patients taken to TLH were divided into 2 groups (treatment group, cefazolin 2 g intravenous single dose before surgery + metronidazole vaginal ovules for 5 days postoperatively, control group: cefazolin 2 g intravenous single dose + placebo vaginal ovules for 5 days postoperatively). MEASUREMENTS AND MAIN RESULTS: The absolute frequency (AF) of PC and PA and their relationship with the presence of bacterial vaginosis (BV) were measured. There was no difference in AF of PC (AF, 2/285 [0.7%] vs 5/284 [1.7%] in the treatment and placebo groups, respectively; risk ratio, 1.75; 95% confidence interval, 0.54-5.65; p = .261), nor for PA (AF, 0/285 [0%] vs 2/289 [0.7%]; p = .159, in the treatment and placebo groups, respectively). The incidence of BV was higher in the metronidazole group than the placebo group (42.5% vs 33.4%, p = .026). CONCLUSION: The use of vaginal metronidazole ovules during the first 5 days in postoperative TLH added to conventional cefazolin prophylaxis does not prevent the development of PC or PA, regardless of the patient's diagnosis of BV.