A high-throughput method for quantifying Drosophila fecundity.

Measurements of Drosophila fecundity are used in a wide variety of studies, such as investigations of stem cell biology, nutrition, behavior, and toxicology. In addition, because fecundity assays are performed on live flies, they are suitable for longitudinal studies such as investigations of aging or prolonged chemical exposure. However, standard Drosophila fecundity assays have been difficult to perform in a high-throughput manner because experimental factors such as the physiological state of the flies and environmental cues must be carefully controlled to achieve consistent results. In addition, exposing flies to a large number of different experimental conditions (such as chemical additives in the diet) and manually counting the number of eggs laid to determine the impact on fecundity is time-consuming. We have overcome these challenges by combining a new multiwell fly culture strategy with a novel 3D-printed fly transfer device to rapidly and accurately transfer flies from one plate to another; the RoboCam, a low-cost, custom built robotic camera to capture images of the wells automatically; and an image segmentation pipeline to automatically identify and quantify eggs. We show that this method is compatible with robust and consistent egg laying throughout the assay period; and demonstrate that the automated pipeline for quantifying fecundity is very accurate (r2 = 0.98 for the correlation between the automated egg counts and the ground truth) In addition, we show that this method can be used to efficiently detect the effects on fecundity induced by dietary exposure to chemicals. Taken together, this strategy substantially increases the efficiency and reproducibility of high throughput egg laying assays that require exposing flies to multiple different media conditions.

The Alignment of the Tibial Component in Total Knee Arthroplasty: Is a Technology-Assisted System Better Than Conventional Instrumentation?

Background Technologies such as navigation and robotics are aimed at improving tibial alignment in total knee arthroplasties (TKA) and eliminating the errors resulting from the use of manual instrumentation. Methods This prospective study analyzed 130 arthroplasties in order to determine whether navigation can improve the frontal mechanical axis of the tibia and whether the postoperative angulation of this axis differs from the preoperative one. The mean patient age was 71.8 years, and the mean BMI was 31.17. Eighty-six patients were female. The same cemented TKA model and the same imageless navigation system were used in all cases. Results The mean postoperative tibial angle following implantation was 87.65°, without any statistically significant differences with respect to the previous angulation. However, navigation was seen to result in a nearly neutral tibial axis, a larger number of cases (41.5%-60.8% {p = 0.002}) aligned within the safe zone (90° ± 3°), a smaller number of outliers, and a clustering of values around the mean. Conclusions Navigation improves the frontal positioning of the tibial component in total knee arthroplasties but does not offer any advantages as compared with conventional instrumentation.

Significance of clinical-immunological patterns and diagnostic yield of biopsies in microscopic polyangiitis and granulomatosis with polyangiitis.

BACKGROUND: Microscopic polyangiitis (MPA) and granulomatosis with polyangiitis (GPA) are the two major antineutrophil cytoplasmic antibody (ANCA)-associated vasculitis (AAV). OBJECTIVES: To characterize a homogenous AAV cohort and to assess the impact of clinicopathological profiles and ANCA serotypes on clinical presentation and prognosis. Clinical differences in GPA patients according to ANCA serotype and the diagnostic yield for vasculitis of biopsies in different territories were also investigated. RESULTS: This retrospective study (2000-2021) included 152 patients with AAV (77 MPA/75 GPA). MPA patients (96.1% myeloperoxidase [MPO]-ANCA and 2.6% proteinase 3 [PR3]-ANCA) presented more often with weight loss, myalgia, renal involvement, interstitial lung disease (ILD), cutaneous purpura, and peripheral nerve involvement. Patients with GPA (44% PR3-ANCA, 33.3% MPO, and 22.7% negative/atypical ANCA) presented more commonly with ear, nose, and throat and eye/orbital manifestations, more relapses, and higher survival than patients with MPA. GPA was the only independent risk factor for relapse. Poor survival predictors were older age at diagnosis and peripheral nerve involvement. ANCA serotypes differentiated clinical features in a lesser degree than clinical phenotypes. A mean of 1.5 biopsies were performed in 93.4% of patients in different territories. Overall, vasculitis was identified in 80.3% (97.3% in MPA and 61.8% in GPA) of patients. CONCLUSIONS: The identification of GPA presentations associated with MPO-ANCA and awareness of risk factors for relapse and mortality are important to guide proper therapeutic strategies in AAV patients. Biopsies of different affected territories should be pursued in difficult-to-diagnose patients based on their significant diagnostic yield.

Gorlin-Like Phenotype in a Young Girl With a De Novo PTCH2 Variant Mutation of Uncertain Significance.

ABSTRACT: Gorlin syndrome, also known as basal cell nevus syndrome, is an autosomal dominant genetic disorder that predisposes humans to tumors. In most cases, this syndrome results from inactivating mutations in the patched homologue 1 gene. Basal cell carcinomas are one of the main characteristics of this syndrome and serve as a major diagnostic criterion. Gorlin syndrome shows a variable phenotype, and recently, other less common mutations in the suppressor of fused homologue or patched homologue 2 genes have been documented in individuals with this syndrome. We present the case of a patient with early-onset basal cell carcinomas and a mild Gorlin syndrome phenotype, attributed to a de novo patched homologue 2 variant of uncertain significance, which has not been previously reported in the literature.

Actinic conjunctivitis and its relationship with prurigo in an indigenous population from the Highlands of Chiapas, Mexico.

PURPOSE: Actinic conjunctivitis (AC), along with cheilitis (AChe), is part of the clinical spectrum of actinic prurigo (AP), a rare photo dermatosis that affects high-risk populations. We analyzed the clinical manifestations and onset of actinic conjunctivitis (AC), and its relationship with prurigo (AP) in a susceptible population. METHODS: This prospective observational cohort study was performed on Indigenous populations from the highlands of Chiapas, Mexico. Thorough dermatological and ophthalmological examinations were performed in patients attending a primary health care center. The clinical features, labor and environmental factors, onset timing, and clinical staging of AC and AP were analyzed. RESULTS: Of the 2913 patients studied, 54 patients (108 eyes) (1.8%) had AC, and 14 patients (25.9%) had AP. The mean age at diagnosis was 36.18 ± 18.52 years (6-70 years). The mean residential altitude was 1884 ± 434.2 m above sea level. Mean self-reported sun exposure was 5.14 ± 3.1 h a day (0.5-12 h). A total of 90.7% reported exposure to biomass fuels during cooking, and 50% to farm animals. AC was the sole manifestation in 70% of the cases. All patients had nasal and temporal photo-exposed conjunctiva. Among the eyes, 12.9% were classified as stage-1, 64.8% as stage-2, and 22.2% stage-3. A total of 83.3% of the patients had hyperpigmented lesions, and 35.1% had evaporative dry eye disease. CONCLUSIONS: AC may be the initial or sole manifestation of AP. Most AC cases (87%) were initially observed at the advanced stages of the disease. Although solar exposure was not associated with late AC stages, a positive association was found with farm animal exposure. Evaporative dry eye associated with meibomian gland dysfunction has not been previously reported in patients with AC.

Psychosocial and vocational impact of an internship program for people diagnosed with severe mental illnesses.

PURPOSE: Unemployment is an obstacle in recovery for people with severe mental illnesses (SMI), both in reducing psychopathological symptoms and in achieving a high quality of life. The aim of this study was to explore the impact of an internship program for people diagnosed with SMI on psychosocial variables using a pre-test and post-test design. METHOD: Four scales were administered to a sample of 127 study participants, all of whom were diagnosed with SMI. The results of standard and sheltered employment outcomes were obtained by comparing the number and type of contracts gained by participants up to one year after program completion. A repeated measures ANOVA analysis was carried out with intervention being an independent intra-subject variable and type of participant diagnosis being a between-subjects variable. RESULTS: A limited increase in social functioning was found, as well as a significant decrease in psychiatric distress for users diagnosed with personality disorders. Participants in the internship program obtained a greater number of employment contracts than their peers who did not participate in the program, albeit with a low effect size. CONCLUSIONS: Though results were positive, extension of internship length and provision of more intensive counselling for participants may be useful.

An internship program for persons with severe mental illnesses yielded modest improvements in social functioning.A decrease in psychiatric distress in participants with personality disorders was found after the internships.Participants in the internship obtained a greater number of employment contracts.An extended internship programme with workplace support is likely to be beneficial.

Comparative analysis of patellar tendon, achilles tendon and plantar fascia structure in indoor and outdoor football players: a novel cross-sectional pilot study.

Different sport modalities were associate with tendon adaptation or even tendon disturbances, such as volleyball, soccer or basketball. Purpose: the aim of the present study was to determine de difference between indoor and outdoor football players on patellar tendon (PT), Achilles tendon (AT), plantar fascia (FP) and Hoffa's fat pad thickness assessed with ultrasound imaging (USI). A cross-sectional study was developed with a total sample of 30 soccer players divided in two groups: outdoor group (n = 15) and indoor group (n = 15). The thickness of PT, AT, PF and Hoffa's fat pad has been assessed with USI. Hoffa's fat pad reported significant differences for the left side between groups (P = 0.026). The rest of variables did not show any significant difference (P < 0.05). The ultrasonography assessment of the thickness of the PT, AT and PF did not show differences between outdoor and indoor football players. Hoffa's fat pad resulted showed a significant decrease for outdoor soccer players with respect futsal players. Thus, it can be considered that the load stimuli received in both soccer players were not enough to produce structural adaptations in PT, AT and PF tissues.

Maternal and fetal outcomes of pregnancy in women with primary systemic vasculitis: A single-center cohort study of 20 patients and 30 pregnancies.

OBJECTIVES: To analyze pregnancy outcomes of patients with primary systemic vasculitis followed in a third-level referral center. METHODS: Retrospective cohort study of all pregnant women with systemic vasculitis followed between 2009 and 2022 at the High-Risk Pregnancy Clinic of the Department of Systemic Autoimmune Diseases of the Hospital Clínic, Barcelona. RESULTS: Twenty women with primary vasculitis were identified, with a total of 30 pregnancies. Antineutrophil cytoplasmic antibody (ANCA)-associated vasculitis (n = 7) and Behçet disease (n = 4) were the most frequent types of vasculitis. All women had the diagnosis of vasculitis before pregnancy, with a median time between disease diagnosis and pregnancy of 5.8 years (range: 2 months-29 years). Most were in remission at conception (76.7 %). During pregnancy, a vasculitis flare occurred in 4 (13.3 %) patients (one each with Takayasu arteritis, eosinophilic granulomatosis with polyangiitis [EGPA], IgA vasculitis [IgAV], and Behçet disease [BD]). Four (16.7 %) of the successful pregnancies had post-partum relapses (one each with EGPA, granulomatosis with polyangiitis, IgAV, and BD). Eighty percent of pregnancies resulted in live babies. In four cases (13.3 %), medical termination of pregnancy was decided, considering the mother or baby health risk. There were two spontaneous miscarriages, and no stillbirths or neonatal deaths. Preeclampsia was the most frequent maternal complication (25 %). Newborns were preterm in 24 % and low birthweight in 20 % of cases. No maternal deaths occurred. CONCLUSIONS: This cohort study shows that vasculitis relapses during pregnancy and post-partum, together with other pregnancy complications, occur in a considerable number of patients with systemic vasculitides, although a final good pregnancy outcome can be expected in most cases. These findings emphasize the convenience of managing these special situations in expert reference centers.

Self-Diagnosis of SARS-CoV-2 from Saliva Samples at Home: Isothermal Amplification Enabled by Do-It-Yourself Portable Incubators and Laminated Poly-ethyl Sulfonate Membranes.

COVID-19 made explicit the need for rethinking the way in which we conduct testing for epidemic emergencies. During the COVID-19 pandemic, the dependence on centralized lab facilities and resource-intensive methodologies (e.g., RT-qPCR methods) greatly limited the deployment of widespread testing efforts in many developed and underdeveloped countries. Here, we illustrate the development of a simple and portable diagnostic kit that enables self-diagnosis of COVID-19 at home from saliva samples. We describe the development of a do-it-yourself (DIY) incubator for Eppendorf tubes that can be used to conduct SARS-CoV-2 detection with competitive sensitivity and selectivity from saliva at home. In a proof-of-concept experiment, we assembled Eppendorf-tube incubators at our home shop, prepared a single-tube mix of reagents and LAMP primers in our lab, and deployed these COVID-19 detection kits using urban delivery systems (i.e., Rappifavor or Uber) to more than 15 different locations in Monterrey, México. This straightforward strategy enabled rapid and cost-effective at-home molecular diagnostics of SARS-CoV-2 from real saliva samples with a high sensitivity (100%) and high selectivity (87%).

Wet anaerobic digestion of organic fraction of municipal solid waste: experience with long-term pilot plant operation and industrial scale-up.

This paper presents the analysis of a pilot anaerobic digestion plant that operates with organic fraction of municipal solid waste (OFMSW) from a wholesale market and can treat up to 500 kg d-1. The process was monitored for a period of 524 days during which the residue was characterized and the biogas production and methane content were recorded. The organic load rate (OLR) of volatile solids (VS) was 0.89 kg m-3 d-1 and the Hydraulic Retention Time (HRT) was 25 d during the process. The yield was 82 Nm3 tons OFMSW-1 biogas, equivalent to 586 Nm3 tons CH4 VS-1. The results obtained in the pilot plant were used to carry out a technical-economic evaluation of a plant that treats 50 tons of OFMSW from wholesale markets. A production of 3769 Nm3 d-1 of biogas and 2080 Nm3 d-1 of methane is estimated, generating 35.1 MWh d-1 when converted to electricity.

Structural, biochemical and immunochemical characterization of an acidic phospholipase A2 from Lachesis acrochorda (Viperidae: Crotalinae) venom.

Viperids of the genus Lachesis, also known as bushmasters, are capable of injecting great amounts of venom that cause severe envenomation incidents. Since phospholipases type A2 are mainly involved in edema and myonecrosis within the snakebite sites, in this work, the isolation, amino acid sequence and biochemical characterization of the first phospholipase type A2 from the venom of Lachesis acrochorda, named Lacro_PLA2, is described. Lacro_PLA2 is an acidic aspartic 49 calcium-dependent phospholipase A2 with 93% similarity to the L. stenophrys phospholipase. Lacro_PLA2 has a molecular mass of 13,969.7 Da and an experimental isoelectric point around 5.3. A combination of N-terminal Edman degradation and MS/MS spectrometry analyses revealed that Lacro_PLA2 contains 122 residues including 14 cysteines that form 7 disulfide bridges. A predicted 3D model shows a high resemblance to other viperid phospholipases. Nevertheless, immunochemical and phospholipase neutralization tests revealed a notorious level of immunorecognition of the isolated protein by two polyclonal antibodies from viperids from different genus, which suggest that Lacro_PLA2 resembles more to bothropic phospholipases. Lacro_PLA2 also showed significantly high edema activity when was injected into mice; so, it could be an alternative antigen in the development of antibodies against toxins of this group of viperids, seeking to improve commercial polyclonal antivenoms.

Retrospective evaluation of hematological ratios in canine parvovirosis: 401 cases.

BACKGROUND: The utility of neutrophil-to-lymphocyte ratio (NLR), platelet to-lymphocyte ratio (PLR) and monocyte-to-lymphocyte ratio (MLR) as prognostic indicators has not been investigated in canine parvovirosis (CPV). HYPOTHESIS: To evaluate whether these hematological ratios obtained at hospital admission in CPV are associated with outcome or duration of hospitalization. ANIMALS: Four hundred one client-owned dogs presented with CPV. Methods-Retrospective multicenter cohort study. Medical records were reviewed to identify dogs with CPV. Data regarding signalment, complete blood count at admission, duration of hospitalization and outcome were collected. RESULTS: Of the 401 dogs included in the study, 336 (83.8%) survived to discharge. The median (25th and 75th percentiles) PLR in nonsurvivors (336.56 [159.84-635.77]) was significantly higher than in survivors (217.65 [117.67-389.65]) (P = .003). The area under the receiver-operating characteristic curve for nonsurvival was 0.615 (95% CI [0.593-0.691], P = .003). A cut off of 700 showed a 21.5% sensitivity and 90% specificity for nonsurvival. No association was observed between hospitalization duration and either hematological ratios or total WBC counts. The median (25th and 75th percentiles) lymphocyte count was below reference interval in all dogs and was significantly lower in the dogs which died (0.82 × 109 /L [0.5-1.87]) than in survivors (1.27 × 109 /L [0.73-2.22]) (P = .005). The median (25th and 75th percentiles) monocyte count however was lower in survivors (0.38 × 109 /L [0.29-1.59]), than in nonsurvivors (0.73 × 109 /L [0.1-2]) (P = .002). CONCLUSIONS: Evaluation of PLR at hospital admission might be a useful marker of disease severity and could have prognostic value in dogs with CPV.

Análisis de los factores pronósticos de mortalidad en los pacientes con enfermedad renal crónica en hemodiálisis

Introducción: La enfermedad renal crónica es una de las principales causas de mortalidad en todo el mundo. La estratificación del riesgo a través del análisis de factores pronósticos podría generar un cambio de paradigma. Objetivo: Analizar los factores pronósticos de mortalidad en los pacientes con enfermedad renal crónica en hemodiálisis. Métodos: Se realizó un estudio no experimental, longitudinal de cohorte retrospectivo en los pacientes con enfermedad renal crónica en hemodiálisis en el Hospital General Docente: "Dr. Ernesto Guevara de la Serna" durante el período del 1 de enero de 2017 al 31 de diciembre de 2021. En general, se analizaron los factores pronósticos de mortalidad mediante el análisis multivariado de regresión logística binaria y se determinó el porcentaje correcto de clasificación del modelo de regresión. Resultados: Se analizaron como variables pronosticas de mortalidad la enfermedad cardiovascular [B = 3,831; p = 0,000; Exp (B) = 46,118], Albúmina 17 mmol/L [B = 1,326; p = 0,027; Exp (B) = 3,767], glucemia < 4 mmol/L [B = 1,600; p = 0,015; Exp (B) = 4,955] y ganancia de peso interdialítica excesiva [B = 2,243; p = 0,001; Exp (B) = 9,420]. El porcentaje global de clasificación del modelo de regresión logística binaria fue de 89,5 %. Conclusiones: Se analizó el modelo predictivo de regresión logística que presentó una buena precisión con los factores de pronósticos asociados a la mortalidad en los pacientes en hemodiálisis.

Introduction: Chronic kidney disease is one of the main causes of mortality worldwide. Risk stratification through the analysis of prognostic factors could generate a paradigm shift. Objective: To analyze the prognostic factors of mortality in patients with chronic kidney disease on hemodialysis. Methods: A non-experimental, longitudinal retrospective cohort study was carried out on patients with chronic kidney disease on hemodialysis at Dr. Ernesto Guevara de la Serna General Teaching Hospital from January 2017 to December 31, 2021. The prognostic factors of mortality were analyzed using multivariate binary logistic regression analysis and the correct percentage of classification of the regression model was determined. Results: Prognostic variables of mortality were analyzed, such as cardiovascular disease [B = 3.831; p = 0.000; Exp (B) = 46.118], albumin 17 mmol/L [B = 1.326; p = 0.027; Exp (B) = 3.767], blood glucose < 4 mmol/L [B = 1.600; p = 0.015; Exp (B) = 4.955] and excessive interdialytic weight gain [B = 2.243; p = 0.001; Exp(B) = 9.420]. The overall classification percentage of the binary logistic regression model was 89.5%. Conclusions: The logistic regression predictive model was analyzed, which showed good precision with the prognostic factors associated with mortality in hemodialysis patients.

Acute and Chronic Effects of Blood Flow Restriction Training in Physically Active Patients With Anterior Cruciate Ligament Reconstruction: A Systematic Review.

CONTEXT: Muscle atrophy and loss of knee function are common findings after anterior cruciate ligament (ACL) reconstruction. Rehabilitation through blood flow restriction (BFR) has gained clinical relevance when combined with low loads to improve these disorders in recent years. OBJECTIVE: To evaluate the rehabilitation effectiveness of ACL reconstruction with the use of BFR on pain, functionality, strength, and muscle mass in physically active people. DATA SOURCES: A search of PubMed, Web of Science, and MEDLINE was performed on March 31, 2023, following the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) 2020 guidelines. STUDY SELECTION: Randomized clinical trials with active adults who underwent ACL surgery were included. They had to compare conventional treatments with the use of BFR, reporting values of pain, functionality, strength, or cross-sectional area (CSA). Articles whose participants presented concomitant injuries and whose intervention combined the use of BFR with treatments other than resistance training were excluded. STUDY DESIGN: Systematic review. LEVEL OF EVIDENCE: Level 2. DATA EXTRACTION: Study design, population, cuff pressure, and main outcomes including strength, quadriceps CSA, pain, and functionality. RESULTS: Six studies out of a total of 389 were included (152 participants; 90 men and 62 women). These included studies showed no differences on CSA or strength when comparing BFR training with high loads exercise. BFR has demonstrated improvements in knee functionality and pain compared with other interventions such as immobilization or high loads training. CONCLUSION: The use of low loads combined with BFR improves pain, strength, functionality, and CSA. In addition, knee pain reduction and functionality are greater with BFR compared with the use of high loads or immobilization.

Haemato-biochemical characterization of equine piroplasmosis asymptomatic carriers and seropositive, real-time PCR negative horses.

Equine piroplasmosis (EP) is caused by Theileria equi and Babesia caballi, transmitted by tick vectors. Horses can suffer an acute, subacute, and chronic forms of the disease, with clinical signs such as poor performance, fever, pale mucosal membranes, and jaundice. The diagnosis of EP subclinical cases is complex due to the sensitivity of real-time PCR and the limited parasite load in some carriers, making it challenging to differentiate them from seropositive, PCR negative (S+PCR-) individuals. This study aimed to describe haematological and biochemical changes in asymptomatic EP carriers, EP S+PCR- horses and control horses (EP seronegative and PCR negative). It also investigated potential haemato-biochemical markers to aid in distinguishing true EP carriers alongside molecular and serological tests. A comprehensive haematology and biochemistry profile was conducted on 410 sera and EDTA blood samples, comprising 130 EP positives by real-time PCR and competitive ELISA (cELISA) (carriers), 130 EP negatives by real-time PCR but positive to cELISA (S+PCR-) and 150 EP negative horses to real-time PCR and c-ELISA (controls). Our study confirmed that a haematological and biochemistry profile could help to differentiate between EP carriers/S+PCR- from healthy horses. Carriers and S+PCR- horses showed significant increases in the white blood cell count (WBC), high total proteins (TP) and total globulins (GLOB) concentration, and liver function markers compared to controls. Additionally, the evaluation of uric acid (UA) suggested oxidative stress in carrier horses. However, no useful haemato-biochemical diagnostic markers were identified to aid the challenging differentiation of EP carriers and S+PCR- horses, highlighting the need for improvement in molecular/serological diagnosis for these horses.

In inflammatory myopathies, dropped head/bent spine syndrome is associated with scleromyositis: an international case-control study.

BACKGROUND: Some myopathies can lead to dropped head or bent spine syndrome (DH/BS). The significance of this symptom has not been studied in inflammatory myopathies (IM). OBJECTIVES: To assess the significance of DH/BS in patients with IM. METHODS: Practitioners from five IM networks were invited to report patients with IM suffering from DH/BS (without other known cause than IM). IM patients without DH/BS, randomly selected in each participating centre, were included as controls at a ratio of 2 to 1. RESULTS: 49 DH/BS-IM patients (DH: 57.1%, BS: 42.9%) were compared with 98 control-IM patients. DH/BS-IM patients were older (65 years vs 53 years, p<0.0001) and the diagnosis of IM was delayed (6 months vs 3 months, p=0.009). Weakness prevailing in the upper limbs (42.9% vs 15.3%), dysphagia (57.1% vs 25.5%), muscle atrophy (65.3% vs 34.7%), weight loss (61.2% vs 23.5%) and loss of the ability to walk (24.5% vs 5.1%) were hallmarks of DH/BS-IM (p≤0.0005), for which the patients more frequently received intravenous immunoglobulins (65.3% vs 34.7%, p=0.0004). Moreover, DH/BS-IM patients frequently featured signs and/or complications of systemic sclerosis (SSc), fulfilling the American College of Rheumatology/European Alliance of Associations for Rheumatology criteria for this disease in 40.8% of the cases (vs 5.1%, p<0.0001). Distribution of the myopathy, its severity and its association with SSc were independently associated with DH/BS (p<0.05). Mortality was higher in the DH/BS-IM patients and loss of walking ability was independently associated with survival (p<0.05). CONCLUSION: In IM patients, DH/BS is a marker of severity and is associated with SSc (scleromyositis).