Longitudinal outcomes of obeticholic acid therapy in ursodiol-nonresponsive primary biliary cholangitis: Stratifying the impact of add-on fibrates in real-world practice.

BACKGROUND: Suboptimal response to ursodeoxycholic acid occurs in 40% of primary biliary cholangitis (PBC) patients, affecting survival. Achieving a deep response (normalisation of alkaline phosphatase [ALP] and bilirubin ≤0.6 upper limit of normal) improves survival. Yet, the long-term effectiveness of second-line treatments remains uncertain. AIMS: To evaluate the long-term effectiveness of obeticholic acid (OCA) ± fibrates. Focusing on biochemical response (ALP ≤1.67 times the upper limit of normal, with a decrease of at least 15% from baseline and normal bilirubin levels), normalisation of ALP, deep response and biochemical remission (deep response plus aminotransferase normalisation). METHODS: We conducted a longitudinal, observational, multicentre study involving ursodeoxyccholic acid non-responsive PBC patients (Paris-II criteria) from Spain and Portugal who received OCA ± fibrates. RESULTS: Of 255 patients, median follow-up was 35.1 months (IQR: 20.2-53). The biochemical response in the whole cohort was 47.2%, 61.4% and 68.6% at 12, 24 and 36 months. GLOBE-PBC and 5-year UK-PBC scores improved (p < 0.001). Triple therapy (ursodeoxycholic acid plus OCA plus fibrates) had significantly higher response rates than dual therapy (p = 0.001), including ALP normalisation, deep response and biochemical remission (p < 0.001). In multivariate analysis, triple therapy remained independently associated with biochemical response (p = 0.024), alkaline phosphatase normalisation, deep response and biochemical remission (p < 0.001). Adverse effects occurred in 41.2% of cases, leading to 18.8% discontinuing OCA. Out of 55 patients with cirrhosis, 12 developed decompensation. All with baseline portal hypertension. CONCLUSION: Triple therapy was superior in achieving therapeutic goals in UDCA-nonresponsive PBC. Decompensation was linked to pre-existing portal hypertension.

Glycerol is not suitable for the cryopreservation of quail semen.

1. This study investigated the potential use of glycerol as a cryoprotectant for quail sperm cells. Its role in maintaining sperm fertilising ability in vivo and in vitro quality parameters, such as motility, was assessed.2. The data showed that the presence of glycerol in semen samples was associated with infertility, which suggested that removal prior to insemination is mandatory. Removal through serial dilution centrifugation was associated with fewer than 5% of motile sperm cells and resulted in no fertility.3. In conclusion, glycerol alone is not suitable for quail semen cryopreservation, and other approaches need to be investigated to develop cryobanking programmes for this species.

Morphometric evaluation of the linea alba in fresh corpses.

Purpose: Diastasis recti abdominis is an increase in the distance between the medial borders of the two rectus muscles. It is most often triggered after intra-abdominal pressure increases, such as postpartum or in obesity. Most publications are based on radiological studies or are done in certain subgroups, without unanimous reference values of the distance between the rectus abdominis or standardization. Methods: Forty-one cadavers were studied. Exclusion criteria: signs of abdominal trauma, major burns, presence of scar from previous abdominal surgery, clinical signs of abdominal hernia, and identification of hernia during cadaver dissection. Linea alba (LA) length, width, and thickness were measured with a flexible tape measure and digital caliper. Anatomical landmarks were established, and subdivisions were described based on them to compare the cadavers. Results: Sex and age had little effect on LA width, thickness, or length. Obesity (compared to normal weight) was the only variable that promoted an increase in the LA width (p < 0.01). The supraumbilical length varied with the total height of the evaluated cadavers (p < 0.01), but the infraumbilical length did not (p = 0.11). Conclusion: The general statistical results of this study, regarding the evaluation of LA measurements in cadavers, showed that ethnicity, sex, and age have little effect on the width, thickness, or length of the LA. LA width differed significantly with abdominal circumference.

Estrangulated obturatory hernia. Case report of a challenging diagnosis of obstructive syndrome.

INTRODUCTION AND IMPORTANCE: Obturator hernia accounts for less than 1 % of all documented cases of pelvic hernias. It most commonly presents as an obstructive syndrome in elderly, multiparous, slim women, characterized by a wider pelvis that facilitates the passage of the hernia sac through the obturator foramen alongside the obturator nerve. In this case, adhering to the SCARE (Updating Consensus Surgical CAse REport) checklist criteria, we present a typical scenario involving an elderly woman who was initially misdiagnosed with a fecaloma, concealing an obturator hernia. CASE REPORT: An 85-year-old patient, displaying prodromal signs of senile disease, presented for medical attention with incapacitating abdominal pain in the right iliac fossa, accompanied by nausea and vomiting. Rectal examination revealed the presence of a fecaloma, and glycerin administration was performed rectally. The patient's condition worsened with the development of mental confusion and hyperactive delirium. Abdominal Computer tomography scan (CT scan) revealed right obturator hernia with enteral segment insinuation and dilation of the proximal bowel. An infraumbilical laparotomy was performed. The herniation of an ileal segment and the right ovary through the obturator foramen was identified. The content proved irreducible to manual maneuvers, leading to obturator muscle section following the dissection of the Retzius space. The right round ligament of the uterus was sectioned, and we manage to preserve the Obturatory branch of the lumbar plexus throughout dissection. A polypropylene mesh was positioned and secured with non-absorbable sutures on the Cooper's ligament, iliac crest, and obturator muscle and segmental enterectomy with primary anastomosis using a linear stapler was performed. CLINICAL DISCUSSION: This demanding case brings to the spotlight the importance of reevaluating even the usual cases. We provide our experience bringing together an unusual diagnosis after the conduction of a once diagnosed fecaloma that almost went down to a perforated acute abdomen. Hence the importance of suspect obturator hernia in unknown obstructive abdomen in elderly women. CONCLUSION: With this report we aim to raise awareness of careful propaedeutic inquiry of acute abdomen. We provide our experience bringing together the diagnosis that agrees with our literature review. Elderly patients commonly challenge the clinical evaluation, especially those with signs of senile disease. Thereby, inkling hidden diagnosis in typical scenarios can improve the patient's care in emergency settings.

Maternal environmental enrichment protects neonatal brains from hypoxic-ischemic challenge by mitigating brain energetic dysfunction and modulating glial cell responses.

There is evidence that maternal milieu and changes in environmental factors during the prenatal period may exert a lasting impact on the brain health of the newborn, even in case of neonatal brain hypoxia-ischemia (HI). The present study aimed to investigate the effects of maternal environmental enrichment (EE) on HI-induced energetic and metabolic failure, along with subsequent neural cell responses in the early postnatal period. Male Wistar pups born to dams exposed to maternal EE or standard conditions (SC) were randomly divided into Sham-SC, HI-SC, Sham-EE, and HI-EE groups. Neonatal HI was induced on postnatal day (PND) 3. The Na+,K+-ATPase activity, mitochondrial function and neuroinflammatory related-proteins were assessed at 24 h and 48 h after HI. MicroPET-FDG scans were used to measure glucose uptake at three time points: 24 h post-HI, PND18, and PND24. Moreover, neuronal preservation and glial cell responses were evaluated at PND18. After HI, animals exposed to maternal EE showed an increase in Na+,K+-ATPase activity, preservation of mitochondrial potential/mass ratio, and a reduction in mitochondrial swelling. Glucose uptake was preserved in HI-EE animals from PND18 onwards. Maternal EE attenuated HI-induced cell degeneration, white matter injury, and reduced astrocyte immunofluorescence. Moreover, the HI-EE group exhibited elevated levels of IL-10 and a reduction in Iba-1 positive cells. Data suggested that the regulation of AKT/ERK1/2 signaling pathways could be involved in the effects of maternal EE. This study evidenced that antenatal environmental stimuli could promote bioenergetic and neural resilience in the offspring against early HI damage, supporting the translational value of pregnancy-focused environmental treatments.

Production of a Ric c3 hypo-allergen with no IgE binding or anaphylactogenic activity.

Several studies have been carried out to expand the use of Ricinus communis L. castor bean (Ricinus communis L castor bean.). This oilseed finds appropriate conditions for its development in Brazil, with more than 700 applications. The main allergens of this plant are Ric c1 and Ric c3, that cross-react with various aeroallergens and food allergens such as peanuts, soybeans, corn, and wheat. This study aimed to determine the effect of mutations in Ric c3 amino acid residues known to affect IgE binding and allergy challenges. Based on the Ric c3 structure, B-cell epitopes, and amino acid involved in IgE binding, we produce recombinant mutant protein, mrRic c3, secreted from E. coli. Strategic glutamic acid residues in IgE-biding regions were changed by Leucine. The allergenicity of mrRic c3 was evaluated by determination of IgE, IgG1, and total IgG in immunized Balb/c mice and by degranulation assays of mast cells isolated from Wistar rats. The mrRic c3 presented a percentage of mast cell degranulation close to that seen in the negative control, and the immunization of mice with mrRic c3 presented lower levels of IgE and IgG1 than the group treated with the protein without mutations. The mutant mrRic c3 had an altered structure and reduced ability to stimulate pro-inflammatory responses and bind IgE but retained its ability to induce blocking antibodies. Thus, producing a hypoallergenic mutant allergen (mrRic c3) may be essential in developing new AIT strategies.

Polymorphisms and haplotypes of TOLLIP and MUC5B are associated with susceptibility and survival in patients with fibrotic hypersensitivity pneumonitis.

INTRODUCTION AND OBJECTIVES: Hypersensitivity pneumonitis (HP) is an interstitial lung disease with diverse clinical features that can present a fibrotic phenotype similar to idiopathic pulmonary fibrosis (IPF) in genetically predisposed individuals. While several single nucleotide polymorphisms (SNPs) have been associated with IPF, the genetic factors contributing to fibrotic HP (fHP) remain poorly understood. This study investigated the association of MUC5B and TOLLIP variants with susceptibility, clinical presentation and survival in Portuguese patients with fHP. MATERIAL AND METHODS: A case-control study was undertaken with 97 fHP patients and 112 controls. Six SNPs residing in the MUC5B and TOLLIP genes and their haplotypes were analyzed. Associations with risk, survival, and clinical, radiographic, and pathological features of fHP were probed through comparisons among patients and controls. RESULTS: MUC5B rs35705950 and three neighboring TOLLIP variants (rs3750920, rs111521887, and rs5743894) were associated with increased susceptibility to fHP. Minor allele frequencies were greater among fHP patients than in controls (40.7% vs 12.1%, P<0.0001; 52.6% vs 40.2%, P = 0.011; 22.7% vs 13.4%, P = 0.013; and 23.2% vs 12.9%, P = 0.006, respectively). Haplotypes formed by these variants were also linked to fHP susceptibility. Moreover, carriers of a specific haplotype (G-T-G-C) had a significant decrease in survival (adjusted hazard ratio 6.92, 95% CI 1.73-27.64, P = 0.006). Additional associations were found between TOLLIP rs111521887 and rs5743894 variants and decreased lung function at baseline, and the MUC5B SNP and radiographic features, further highlighting the influence of genetic factors in fHP. CONCLUSION: These findings suggest that TOLLIP and MUC5B variants and haplotypes may serve as valuable tools for risk assessment and prognosis in fibrotic hypersensitivity pneumonitis, potentially contributing to its patient stratification, and offer insights into the genetic factors influencing the clinical course of the condition.

Sustained PGC-1α2 or PGC-1α3 expression induces astrocyte dysfunction and degeneration.

Peroxisome proliferator-activated receptor-gamma coactivator-1α (PGC-1α) transcriptional coactivators are key regulators of energy metabolism-related genes and are expressed in energy-demanding tissues. There are several PGC-1α variants with different biological functions in different tissues. The brain is one of the tissues where the role of PGC-1α isoforms remains less explored. Here, we used a toxin-based mouse model of Parkinson's disease (PD) and observed that the expression levels of variants PGC-1α2 and PGC-1α3 in the nigrostriatal pathway increases at the onset of dopaminergic cell degeneration. This increase occurs concomitant with an increase in glial fibrillary acidic protein levels. Since PGC-1α coactivators regulate cellular adaptive responses, we hypothesized that they could be involved in the modulation of astrogliosis induced by 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP). Therefore, we analysed the transcriptome of astrocytes transduced with expression vectors encoding PGC-1α1 to 1α4 by massively parallel sequencing (RNA-seq) and identified the main cellular pathways controlled by these isoforms. Interestingly, in reactive astrocytes the inflammatory and antioxidant responses, adhesion, migration, and viability were altered by PGC-1α2 and PGC-1α3, showing that sustained expression of these isoforms induces astrocyte dysfunction and degeneration. This work highlights PGC-1α isoforms as modulators of astrocyte reactivity and as potential therapeutic targets for the treatment of PD and other neurodegenerative disorders.

Diet of crab-eating fox (Cerdocyon thous) in two conservation units of the Amazon rainforest, Brazil

Abstract The present study aimed to evaluate the diet of the free-living crab-eating fox by identifying the stomach contents of the 17 crab-eating foxes (Cerdocyon thous) roadkilled in two conservation units, both located in the Amazon rainforest. The food items were quantified by frequency of occurrence (FO) and percentage of occurrence (PO). The stomach contents were analysed for dry matter (DM), crude protein (CP), crude fibre (CF), ether extract (EE), and mineral matter (MM). Nitrogen-free extractives (NFE), metabolisable energy (ME) values, as well as the energy need for maintenance were estimated. The composition of the diet for the crab-eating fox presented 29 food items from the different taxonomic groups, with a greater diversity of items of animal origin (n=22), although the highest frequency of occurrence was gramineae (Poaceae) (41.18%). Among the items of animal origin, 21% were mammals, 18% reptiles, 10% amphibians, 9% invertebrates and 3% birds. A high content of CF (62.76%) were determined. Nitrogen-free extractive and dry matter averages were 5.91% and 141.82 kcal/100g, respectively. The average maintenance energy was 447.01 kcal/day. These findings suggesting that the crab-eating foxes have a generalist diet with an omnivorous diet in the Amazon basin, feeding on gramineae, fruits, insects, snakes, amphibians, birds and small mammals and have the same feeding habit that present in other Brazilian biomes.

Resumo Este estudo objetivou analisar a dieta do cachorro-do-mato, de vida livre, por meio da identificação do conteúdo estomacal de 17 Cerdocyon thous atropelados em duas unidades de conservação da Floresta Amazônica. O conteúdo estomacal foi analisado e os itens alimentares foram quantificados pela frequência de ocorrência (FO) e percentagem de ocorrência (PO). Também foram avaliados os teores de matéria seca (MS), proteína bruta (PB), fibra bruta (FB), extrato etéreo (EE) e, matéria mineral (MM). Foram estimados o extrativo não-nitrogenado (ENN), a energia metabolizável (EM) e a necessidade energética de manutenção. A composição da dieta do cachorro-do-mato apresentou 29 itens alimentares dos diferentes grupos taxonômicos, tendo uma maior diversidade de itens de origem animal (n=22), ainda que a maior FO tenha sido de gramíneas (Poaceae) (41.18%). Dentre os itens de origem animal, 21% eram mamíferos, 18% répteis, 10% anfíbios, 9% invertebrados e 3% aves. Determinou-se um alto teor de FB (62.76%). A média do ENN e da MS foi 5.91% e 141.82 kcal/100g, respectivamente. A média da energia de manutenção foi 447.01 kcal/dia. Estes achados sugerem que o cachorro-do-mato encontrado na Bacia Amazônica é um animal generalista com uma dieta onívora, se alimentando de gramíneas, frutos, insetos, serpentes, anfíbios, aves e pequenos mamíferos, portanto com o mesmo hábito alimentar relatado a esta espécie quando encontrada em outros biomas brasileiros.

Diet of crab-eating fox (Cerdocyon thous) in two conservation units of the Amazon rainforest, Brazil / Dieta do cachorro-do-mato (Cerdocyon thous) em duas unidades de conservação da Floresta Amazônica, Brasil

Abstract The present study aimed to evaluate the diet of the free-living crab-eating fox by identifying the stomach contents of the 17 crab-eating foxes (Cerdocyon thous) roadkilled in two conservation units, both located in the Amazon rainforest. The food items were quantified by frequency of occurrence (FO) and percentage of occurrence (PO). The stomach contents were analysed for dry matter (DM), crude protein (CP), crude fibre (CF), ether extract (EE), and mineral matter (MM). Nitrogen-free extractives (NFE), metabolisable energy (ME) values, as well as the energy need for maintenance were estimated. The composition of the diet for the crab-eating fox presented 29 food items from the different taxonomic groups, with a greater diversity of items of animal origin (n=22), although the highest frequency of occurrence was gramineae (Poaceae) (41.18%). Among the items of animal origin, 21% were mammals, 18% reptiles, 10% amphibians, 9% invertebrates and 3% birds. A high content of CF (62.76%) were determined. Nitrogen-free extractive and dry matter averages were 5.91% and 141.82 kcal/100g, respectively. The average maintenance energy was 447.01 kcal/day. These findings suggesting that the crab-eating foxes have a generalist diet with an omnivorous diet in the Amazon basin, feeding on gramineae, fruits, insects, snakes, amphibians, birds and small mammals and have the same feeding habit that present in other Brazilian biomes.

Resumo Este estudo objetivou analisar a dieta do cachorro-do-mato, de vida livre, por meio da identificação do conteúdo estomacal de 17 Cerdocyon thous atropelados em duas unidades de conservação da Floresta Amazônica. O conteúdo estomacal foi analisado e os itens alimentares foram quantificados pela frequência de ocorrência (FO) e percentagem de ocorrência (PO). Também foram avaliados os teores de matéria seca (MS), proteína bruta (PB), fibra bruta (FB), extrato etéreo (EE) e, matéria mineral (MM). Foram estimados o extrativo não-nitrogenado (ENN), a energia metabolizável (EM) e a necessidade energética de manutenção. A composição da dieta do cachorro-do-mato apresentou 29 itens alimentares dos diferentes grupos taxonômicos, tendo uma maior diversidade de itens de origem animal (n=22), ainda que a maior FO tenha sido de gramíneas (Poaceae) (41.18%). Dentre os itens de origem animal, 21% eram mamíferos, 18% répteis, 10% anfíbios, 9% invertebrados e 3% aves. Determinou-se um alto teor de FB (62.76%). A média do ENN e da MS foi 5.91% e 141.82 kcal/100g, respectivamente. A média da energia de manutenção foi 447.01 kcal/dia. Estes achados sugerem que o cachorro-do-mato encontrado na Bacia Amazônica é um animal generalista com uma dieta onívora, se alimentando de gramíneas, frutos, insetos, serpentes, anfíbios, aves e pequenos mamíferos, portanto com o mesmo hábito alimentar relatado a esta espécie quando encontrada em outros biomas brasileiros.

Cellular dynamics as a marker of normal-to-cancer transition in human cells.

Normal-to-cancer (NTC) transition is known to be closely associated to cell´s biomechanical properties which are dependent on the dynamics of the intracellular medium. This study probes different human cancer cells (breast, prostate and lung), concomitantly to their healthy counterparts, aiming at characterising the dynamical profile of water in distinct cellular locations, for each type of cell, and how it changes between normal and cancer states. An increased plasticity of the cytomatrix is observed upon normal-to-malignant transformation, the lung carcinoma cells displaying the highest flexibility followed by prostate and breast cancers. Also, lung cells show a distinct behaviour relative to breast and prostate, with a higher influence from hydration water motions and localised fast rotations upon NTC transformation. Quasielastic neutron scattering techniques allowed to accurately distinguish the different dynamical processes taking place within these highly heterogeneous cellular systems. The results thus obtained suggest that intracellular water dynamics may be regarded as a specific reporter of the cellular conditions-either healthy or malignant.

Bone anchor as fixation method for abdominal wall reconstruction. A case report about a ten times recurred inguinal hernia.

INTRODUCTION: While the use of bone anchor fixation for abdominal wall reconstruction for supra-pubic incisional hernia is well described (Yee et al., 2008 [1]), we show in this case report, written in line with the SCARE criteria (Agha et al., 2020 [2]), a novel use of this tool as an adjunct in the repair of a ten time recurrent inguinal hernia. CASE REPORT: A 65 years old multiparous, diabetic non-obese female, with previous abdominoplasty was submitted for left inguinal hernia for ten times, between multiples complications between infection, more than one mesh excision by anterior approach and laparoscopic approach. The wide range of procedures culminated in a destruction of the abdominal wall, making it impossible for a usual fixation of mesh in the region. Therefore, a multidisciplinary approach was planned for the patient with a bone anchor as a mesh fixation method. With a year follow up we did not observe a local hernia recurrence. CLINICAL DISCUSSION: Hernia itself is a multifactorial disease. As a anatomical defect, surgery is the only effective treatment. Our report brings a novel approach to a challenging case with many previous unsuccessful applications of conventional surgeries. Hence, we stimulate the multidisciplinary discussion for enhancing post operatory outcomes and a better point of care for the patient.

Strength of a Zirconia- Reinforced Glass-Ceramic After Diamond Bur Adjustment.

INTRODUCTION: Adjustments with diamond burs on the ceramic surface can affect its roughness and decrease flexural strength. This study evaluated the effect of polishing or glazing on the surface roughness and biaxial flexural strength of a zirconia-reinforced lithium silicate glass-ceramic upon adjustment with diamond burs. METHODS: Seventy disks were prepared according to ISO 6872 and divided into seven groups (n = 10) according to different adjustments and finishing processes. Surface roughness was measured prior to biaxial flexural strength testing. The topography was analyzed using an atomic force microscope , fracture markings were identified using a stereomicroscope and representative specimens were analyzed using scanning electron microscopy. RESULTS: The use of diamond burs significantly increased the surface roughness and decreased the strength of the evaluated ceramic (p⟨0.05). Polishing reduced the ceramic's roughness but resulted in flexural strength similar to that of the groups with wear (p⟩0.05). The glaze-treated specimens showed flexural strength statistically similar to the control group (p>0.05) but higher roughness, similar to those with wear. CONCLUSIONS: Polishing reduced the surface roughness but did not affect the biaxial flexural strength of the ZLS ceramic. At the same time, the application of glaze after wear increased the strength.

A far-red-emitting ZnAl1.95Cr0.05O4 phosphor for plant growth LED applications.

ZnAl2-xCrxO4 (x = 0 and 0.05) samples were synthesized via a high-temperature solid-state reaction method. The structure, photoluminescence properties, EPR measurements, thermal stability, and chromaticity diagram of the far-red phosphor ZnAl1.95Cr0.05O4 were investigated. These measurements have enabled us to study the Cr3+ transitions and the site symmetry of Cr3+ in the ZnAl2O4 host lattice and examine the suitability of ZnAl1.95Cr0.05O4 for plant growth application. According to optical and EPR measurements, Cr3+ ions substitute Al3+ ions with D3d symmetry in the ZnAl2O4 host. PLE measurement indicates that upon excitation at 390 nm and 530 nm, the far-red phosphor ZnAl1.95Cr0.05O4 exhibited bright far-red emission around 687 nm. Photoluminescence phenomena show a sharp R line origin from the sublevels of the 2Eg(2G) → 4A2(4F) transition in Cr3+ ions. The 2Eg level was split into 2Eg (Eg) and 2Eg (2Ag) levels in the distorted crystal field environment, and the sharp R line in the ZnAl2O4 matrix was split into R1 and R2 lines. In this paper, the temperature-dependent luminescence characteristics of ZnAl1.95Cr0.05O4 have been investigated. As the temperature increased from 300 K to 440 K, a slight decrease in the intensity of the R1 and R2 lines was observed under excitation at 390 nm. The experimental results show that the ZnAl1.95Cr0.05O4 phosphors exhibit a nearly zero-thermal-quenching behavior. The CIE chromaticity coordinates of the ZnAl1.95Cr0.05O4 phosphor were located at the boundary of the chromaticity diagram, signifying that the phosphors possessed high color purity. The emissions of the ZnAl1.95Cr0.05O4 phosphor match well with the PFR absorption of phytochromes in plants. The investigation indicates that ZnAl1.95Cr0.05O4 is a potential far-red phosphor matching ultraviolet (UV) LED chips for plant growth applications.

Avian sperm-borne RNAs: optimisation of a new isolation protocol.

1. Sperm-borne RNAs are involved in sperm and embryonic protein translation, the regulation of early development and the epigenetic inheritance of the paternal phenotype. Sperm-borne RNA purification protocols generally include a cell purification stage to discard contamination by somatic cells. In avian species, no protocol is currently available to isolate all the populations composing sperm-borne RNAs.2. This study evaluated the presence of somatic cells in semen samples of chickens and quails using visual examination after fluorescent nuclei staining. The efficiency of somatic cell lysis buffer (SCLB) on chicken liver cells and its impacts on chicken sperm cell integrity was explored. Three different approaches were tested to isolate RNA: two developed for mammalian sperm cells and a commercial kit for somatic cells. The efficiency and reliability of each approach was determined based on RNA quality and purity. Eventually, the presence of miRNA and mRNA in purified avian sperm-borne RNAs was investigated by RT-(q)PCR.3. No somatic cells were found in chicken and quail semen. The SCLB totally lysed chicken liver cells but also induced sperm cell necrosis. Consequently, this treatment wasn't performed on samples prior to RNA isolation. Among the tested RNA purification protocols, the commercial one was the least variable and isolated RNA with the highest purity levels. No DNA contamination was observed. Furthermore, the samples contained miRNA and mRNA already known as present in mammalian sperm cells (gga-miR-100-5p, gga-miR-191-5p, GAPDH and PLCZ1), but mRNAs associated with leucocytes (CD4) and Sertoli cells (SOX4, CLDN11) were not detected. This protocol was successfully applied to quail sperm cells.4. Altogether, the study reveals that it is unnecessary to pre-treat samples to remove somatic cell contamination before RNA purification and successfully describes an isolation protocol for sperm-borne RNAs, including small non-coding and long coding RNAs, in two distinct avian species highly valuable as biological models.

Clinical standards for the management of adverse effects during treatment for TB.

BACKGROUND: Adverse effects (AE) to TB treatment cause morbidity, mortality and treatment interruption. The aim of these clinical standards is to encourage best practise for the diagnosis and management of AE.METHODS: 65/81 invited experts participated in a Delphi process using a 5-point Likert scale to score draft standards.RESULTS: We identified eight clinical standards. Each person commencing treatment for TB should: Standard 1, be counselled regarding AE before and during treatment; Standard 2, be evaluated for factors that might increase AE risk with regular review to actively identify and manage these; Standard 3, when AE occur, carefully assessed and possible allergic or hypersensitivity reactions considered; Standard 4, receive appropriate care to minimise morbidity and mortality associated with AE; Standard 5, be restarted on TB drugs after a serious AE according to a standardised protocol that includes active drug safety monitoring. In addition: Standard 6, healthcare workers should be trained on AE including how to counsel people undertaking TB treatment, as well as active AE monitoring and management; Standard 7, there should be active AE monitoring and reporting for all new TB drugs and regimens; and Standard 8, knowledge gaps identified from active AE monitoring should be systematically addressed through clinical research.CONCLUSION: These standards provide a person-centred, consensus-based approach to minimise the impact of AE during TB treatment.

Physical activity measured with an accelerometer in dogs following extracapsular stabilisation to treat cranial cruciate ligament rupture.

OBJECTIVES: To quantify the physical activity levels in dogs with cranial cruciate ligament rupture before and after lateral fabellar suture stabilisation surgery. MATERIALS AND METHODS: Seventeen dogs (mean weight, 12.3±5.1 kg) with unilateral cranial cruciate ligament rupture were fitted with an accelerometer for seven consecutive days at four different time points: before surgery (T0), one (T1), three (T3) and six (T6) months after surgery. The total activity and times spent in sedentary activity, light to moderate activity and vigorous activity were recorded by the accelerometer, and preoperative and postoperative data were compared. At all time points, dogs underwent clinical evaluations (lameness score, stifle pain score and thigh circumference) and their owners were asked to respond to questionnaires to subjectively score the physical activity and quality of life of the dogs. RESULTS: At the four time points, the dogs spent between 21.2 and 21.4 hours on sedentary behaviour, 2.3 and 2.5 hours performing light to moderate activity, and 13 to 15 minutes performing vigorous activity. There was no increase in physical activity variables or decrease in sedentary behaviour over time. Lameness scores, pain score and dogs' quality of life improved significantly during the postoperative period. At T6, 17 (100%) of 17 dogs presented no lameness, 16 (94%) of 17 dogs presented no stifle pain, 16 (94%) of 17 owners rated the quality of life as very good and excellent, and 16 (100%) of 16 owners reported a total return to normal activity levels. CLINICAL SIGNIFICANCE: The clinical recovery after extracapsular stabilisation of the stifle joint was not associated with a spontaneous increase in physical activity or a decrease in sedentary behaviour.

Computationally modelling cell proliferation: A review.

Cell proliferation is vital for the development and homeostasis of the human body. For such to occur, cells go through the cell cycle during which they replicate their genetic material and ultimately complete cellular division, when one cell divides into two new cells with equal genetic material. However, if there are some errors or abnormalities during the cell cycle that disrupt the balance between cell death and proliferation, severe problems can occur, such as tumour development, which is currently one of the leading causes of death in the world. Nowadays, mathematical and computational models are used to understand and study several biological mechanisms and processes, namely cellular proliferation. Over the last forty-five years, several models have attempted to describe cell proliferation and its regulation. Due to the complexity of the process, numerous assumptions and simplifications have been considered. This work presents a review of some of these models, focusing mainly on mammalian or generic eukaryotic models. Previously published continuum, discrete and hybrid approaches are presented and compared, in order to understand and highlight the relevance and capabilities of these models, their shortcomings and future challenges.

Conductive carbon paint: An efficient way to electrodeposit metallic nanowires by using porous membranes.

The present work demonstrates that conductive carbon paint, used for sample preparation in electron microscopy, can be a more straightforward and as-effective substitute for the metallic layer deposition usually used for the electrodeposition of metallic nanowires within porous membranes. AFM images demonstrated the good surface quality of the carbon layer. Raman spectroscopy confirmed the high crystallinity of carbon and high density of π-electrons. The electrical conductivity of the carbon layer was estimated using the linear sweep voltammetry technique. This new cathode was employed to grow continuous (Ni) and composition-modulated (Ni/Cu) nanowires within alumina templates, starting from aqueous solutions of Ni2+ and Cu2+ mixed salts. The obtention of metallic copper and nickel, and their separation can be readily observed by scanning electron microscopy and elemental mapping by EDS.

Carbamazepine, venlafaxine, tramadol, and their main metabolites: Toxicological effects on zebrafish embryos and larvae.

Pharmaceutical compounds and their metabolites are found in natural and wastewater. However, investigation of their toxic effects on aquatic animals has been neglected, especially for metabolites. This work investigated the effects of the main metabolites of carbamazepine, venlafaxine and tramadol. Zebrafish embryos were exposed (0.1-100 µg/L) for 168hpf exposures to each metabolite (carbamazepine-10,11-epoxide, 10,11-dihydrocarbamazepine, O-desmethylvenlafaxine, N-desmethylvenlafaxine, O-desmethyltramadol, N-desmethyltramadol) or the parental compound. A concentration-response relationship was found for the effects of some embryonic malformations. Carbamazepine-10,11-epoxide, O-desmethylvenlafaxine and tramadol elicited the highest malformation rates. All compounds significantly decreased larvae responses on a sensorimotor assay compared to controls. Altered expression was found for most of the 32 tested genes. In particular, abcc1, abcc2, abcg2a, nrf2, pparg and raraa were found to be affected by all three drug groups. For each group, the modelled expression patterns showed differences in expression between parental compounds and metabolites. Potential biomarkers of exposure were identified for the venlafaxine and carbamazepine groups. These results are worrying, indicating that such contamination in aquatic systems may put natural populations at significant risk. Furthermore, metabolites represent a real risk that needs more scrutinising by the scientific community.