Application of real-time PCR for the identification of the endangered species Galemys pyrenaicus through faecal samples.

BACKGROUND: Currently, many micromammals are important targets for study. The endangered Galemys pyrenaicus is an outstanding example. Globally, their populations have suffered a substantial decline in last 20 years. In the surveyed area, the capture of desman is legally forbidden due to the high conservation concerns. Reason by non-invasive sampling through faeces is proposed for its monitoring. Furthermore, the confusion between faeces from desman and Mediterranean water shrews must be considered. Thus, the aim of this study was focused on developing RT-PCR assays to determine the presence of Galemys pyrenaicus and N. a. anomalus from non-invasive samples. METHODS AND RESULTS: The study was conducted in the mountains of the System Central of Extremadura (Spain). A total of 186 samples were collected from 2018 to 2021 by experts where historically reported and/or our previous studies confirmed their presence. RT-PCR assays using hydrolysis probes were designed to detect genetic material from both desman and Mediterranean water shrews and its specificity was confirmed. The reliability of the method was further assessed by PCR sequencing of mitochondrial Cyb and d-loop, resulting fully compatible with the RT-PCR approach. Intraspecific phylogenetic relationship was reported to improve knowledge about mtDNA variability in the desman from the Central System. CONCLUSIONS: We demonstrated that RT-PCR gives a gold opportunity to further map the species using faeces which minimizes disturbance and reports both population status and individual presence. Cost-effective RT-PCR combined with field-collected faeces allows us to better investigate the full range of occurrence of the species.

Non-Invasive Wildlife Disease Surveillance Using Real Time PCR Assays: The Case of the Endangered Galemys pyrenaicus Populations from the Central System Mountains (Extremadura, Spain).

The Iberian desman (Galemys pyrenaicus) is a small semi-aquatic mammal that inhabits mountainous areas from the centre to the north of the Iberian Peninsula and the Pyrenees and is listed as endangered because it has suffered a serious decline. Since 1960, only three species of digeneans (Omphalometra flexuosa, Maritrema pyrenaica and Mathovius galemydis) and two nematodes (Aonchotheca galemydis and Paracuaria hispanica) have been reported from the desman, but no further information on health status and no data from Extremadura has been available. The aim of our study was to characterise the diversity and distribution of parasites and microbiomes of desmans in different areas of the Central System of Extremadura. Between 2019 and 2021 we collected 238 fecal samples and one tissue (intestine) sample that was obtained from a dead desman. DNA templates were processed by commercial or customised real-time PCR using TaqMan probes. Representative data were obtained for Cryptosporidium spp., Omphalometra spp., Eimeria spp., Salmonella spp., Staphylococcus spp. and Leptospira spp. Omphalometra spp. was studied using a newly developed PCR test. The screening of the dead desman allowed us to obtain, for the first time, a partial sequence of the 18SrDNA. This study is the most complete study of the desman, allowing us to identify parasites and the microbiome in populations of G. pyrenaicus using non-invasive sampling.

Resistance to the Bacteriocin Lcn972 Deciphered by Genome Sequencing.

In view of the current threat of antibiotic resistance, new antimicrobials with low risk of resistance development are demanded. Lcn972 is a lactococcal bacteriocin that inhibits septum formation by binding to the cell wall precursor lipid II in Lactococcus. It has a species-specific spectrum of activity, making Lcn972 an attractive template to develop or improve existing antibiotics. The aim of this work was to identify mutations present in the Lcn972-resistant clone Lactococcus cremoris D1-20, previously evolved from the sensitive strain L. cremoris MG1614. Whole-genome sequencing and comparison over the reference genome L. cremoris MG1363 identified several unexpected mutations in the parental strain MG1614, likely selected during in-house propagation. In the Lcn972R clone, two previously identified mutations were mapped and confirmed. Additionally, another transposition event deregulating cellobiose uptake was identified along with three point mutations of unknown consequences for Lcn972 resistance. Two new independent evolution experiments exposing L. cremoris MG1614 to Lcn972 revealed transposition of IS981 into the LLMG_RS12285 locus as the predominant mutation selected by Lcn972. This event occurs early during evolution and was found in 100% of the evolved clones, while other mutations were not selected. Therefore, activation of LLMG_RS12285 coding for a putative anti-ECF (extra-cytoplasmic function) sigma factor is regarded as the main Lcn972 resistance factor in L. cremoris MG1614.

The clinical and genetic spectrum of autosomal-recessive TOR1A-related disorders.

In the field of rare diseases, progress in molecular diagnostics led to the recognition that variants linked to autosomal-dominant neurodegenerative diseases of later onset can, in the context of biallelic inheritance, cause devastating neurodevelopmental disorders and infantile or childhood-onset neurodegeneration. TOR1A-associated arthrogryposis multiplex congenita 5 (AMC5) is a rare neurodevelopmental disorder arising from biallelic variants in TOR1A, a gene that in the heterozygous state is associated with torsion dystonia-1 (DYT1 or DYT-TOR1A), an early-onset dystonia with reduced penetrance. While 15 individuals with AMC5-TOR1A have been reported (less than 10 in detail), a systematic investigation of the full disease-associated spectrum has not been conducted. Here, we assess the clinical, radiological and molecular characteristics of 57 individuals from 40 families with biallelic variants in TOR1A. Median age at last follow-up was 3 years (0-24 years). Most individuals presented with severe congenital flexion contractures (95%) and variable developmental delay (79%). Motor symptoms were reported in 79% and included lower limb spasticity and pyramidal signs, as well as gait disturbances. Facial dysmorphism was an integral part of the phenotype, with key features being a broad/full nasal tip, narrowing of the forehead and full cheeks. Analysis of disease-associated manifestations delineated a phenotypic spectrum ranging from normal cognition and mild gait disturbance to congenital arthrogryposis, global developmental delay, intellectual disability, absent speech and inability to walk. In a subset, the presentation was consistent with foetal akinesia deformation sequence with severe intrauterine abnormalities. Survival was 71%, with higher mortality in males. Death occurred at a median age of 1.2 months (1 week-9 years), due to respiratory failure, cardiac arrest or sepsis. Analysis of brain MRI studies identified non-specific neuroimaging features, including a hypoplastic corpus callosum (72%), foci of signal abnormality in the subcortical and periventricular white matter (55%), diffuse white matter volume loss (45%), mega cisterna magna (36%) and arachnoid cysts (27%). The molecular spectrum included 22 distinct variants, defining a mutational hotspot in the C-terminal domain of the Torsin-1A protein. Genotype-phenotype analysis revealed an association of missense variants in the 3-helix bundle domain to an attenuated phenotype, while missense variants near the Walker A/B motif as well as biallelic truncating variants were linked to early death. In summary, this systematic cross-sectional analysis of a large cohort of individuals with biallelic TOR1A variants across a wide age-range delineates the clinical and genetic spectrum of TOR1A-related autosomal-recessive disease and highlights potential predictors for disease severity and survival.

Mutations, Genes, and Phenotypes Related to Movement Disorders and Ataxias.

Our clinical series comprises 124 patients with movement disorders (MDs) and/or ataxia with cerebellar atrophy (CA), many of them showing signs of neurodegeneration with brain iron accumulation (NBIA). Ten NBIA genes are accepted, although isolated cases compatible with abnormal brain iron deposits are known. The patients were evaluated using standardised clinical assessments of ataxia and MDs. First, NBIA genes were analysed by Sanger sequencing and 59 patients achieved a diagnosis, including the detection of the founder mutation PANK2 p.T528M in Romani people. Then, we used a custom panel MovDisord and/or exome sequencing; 29 cases were solved with a great genetic heterogeneity (34 different mutations in 23 genes). Three patients presented brain iron deposits with Fe-sensitive MRI sequences and mutations in FBXO7, GLB1, and KIF1A, suggesting an NBIA-like phenotype. Eleven patients showed very early-onset ataxia and CA with cortical hyperintensities caused by mutations in ITPR1, KIF1A, SPTBN2, PLA2G6, PMPCA, and PRDX3. The novel variants were investigated by structural modelling, luciferase analysis, transcript/minigenes studies, or immunofluorescence assays. Our findings expand the phenotypes and the genetics of MDs and ataxias with early-onset CA and cortical hyperintensities and highlight that the abnormal brain iron accumulation or early cerebellar gliosis may resembling an NBIA phenotype.

Pearls & Oy-sters: Tumefactive Demyelinating Lesions With MOG Antibodies Preceding Late-Infantile Metachromatic Leukodystrophy.

The development of acute neurological dysfunction associated with tumefactive demyelinating lesions (TDL) and mild diffuse involvement of the corpus callosum has been described in children as a sentinel event that may allow diagnosis of juvenile metachromatic leukodystrophy (MLD) at an early and potentially treatable stage. We report a child that developed this clinico-radiological pattern associated with myelin oligodendrocyte glycoprotein (MOG) antibodies several months prior the onset of progressive symptoms of late infantile MLD.

Comparative Study of Bioactive Lipid Extraction from Squid (Doryteuthis gahi) by-Products by Green Solvents.

A novel approach of bioactive lipid extraction by different green solvents was carried out on squid (Doryteuthis gahi) by-products. By-products (viscera, heads, skin, tails, etc.), considered as a single product, were subjected to the following solvent systems: ethanol, acetone, ethyl acetate, 1/1 ethanol/acetone, 1/1 ethanol/ethyl acetate, and 1/1 acetone/ethyl acetate. Analyses carried out included lipid yield, lipid class content, and fatty acid (FA) composition. Results were compared to the lipid extract obtained by the traditional procedure (1/1 chloroform/methanol). Lipid yields obtained by green solvents led to a 33.4−73.2% recovery compared to traditional extraction; the highest values (p < 0.05) were obtained by ethanol-containing systems. Compared to the traditional procedure, ethanol systems showed an 85.8−90.3% recovery of phospholipid compounds and no differences (p > 0.05) in the ω3/ω6 ratio. Green-extracting systems led to higher average values for eicosapentaenoic acid content (15.66−18.56 g·100 g−1 total FAs) and polyene index (1.93−3.29) than chloroform/methanol extraction; differences were significant (p < 0.05) for systems including acetone and ethyl acetate. No differences (p > 0.05) were detected for docosahexaenoic acid content between the traditional procedure and green systems, with all values being included in the 31.12−32.61 g·100 g−1 total FA range. The suitability of EtOH-containing green systems for extraction of bioactive lipid compounds from squid by-products was concluded.

Impact of COVID-19 in Immunosuppressed Children With Neuroimmunologic Disorders.

BACKGROUND AND OBJECTIVES: To investigate whether children receiving immunosuppressive therapies for neuroimmunologic disorders had (1) increased susceptibility to SARS-CoV2 infection or to develop more severe forms of COVID-19; (2) increased relapses or autoimmune complications if infected; and (3) changes in health care delivery during the pandemic. METHODS: Patients with and without immunosuppressive treatment were recruited to participate in a retrospective survey evaluating the period from March 14, 2020, to March 30, 2021. Demographics, clinical features, type of immunosuppressive treatment, suspected or confirmed COVID-19 in the patients or cohabitants, and changes in care delivery were recorded. RESULTS: One hundred fifty-three children were included: 84 (55%) female, median age 13 years (interquartile range [8-16] years), 79 (52%) on immunosuppressive treatment. COVID-19 was suspected or confirmed in 17 (11%) (all mild), with a frequency similar in patients with and without immunosuppressive treatment (11/79 [14%] vs 6/74 [8%], p = 0.3085). The frequency of neurologic relapses was similar in patients with (18%) and without (21%) COVID-19. Factors associated with COVID-19 included having cohabitants with COVID-19 (p < 0.001) and lower blood levels of vitamin D (p = 0.039). Return to face-to-face schooling or mask type did not influence the risk of infection, although 43(28%) children had contact with a classmate with COVID-19. Clinic visits changed from face to face to remote for 120 (79%) patients; 110 (92%) were satisfied with the change. DISCUSSION: In this cohort of children with neuroimmunologic disorders, the frequency of COVID-19 was low and not affected by immunosuppressive therapies. The main risk factors for developing COVID-19 were having cohabitants with COVID-19 and low vitamin D levels.

Comparison between Tissue Adhesive Cyanoacrylate and Suture for Palatal Fibromucosa Healing: A Randomized Controlled Study.

Cyanoacrylate tissue adhesive is proposed to promote soft tissue healing in oral surgery and minimize complications (pain, inflammation, and bleeding) associated with wound healing by secondary intention. The objective was to compare cyanoacrylate tissue adhesive (test group) with suture (control group) in terms of postoperative complications, operative time, and wound healing in the palatal donor area after harvesting a de-epithelialized gingival graft. A randomized controlled clinical trial was performed in 24 patients randomly assigned to one of two study groups. Data were gathered on wound bleeding, operative time, postoperative pain, inflammation, hyperesthesia, necrosis, and donor area healing time. Operative time was almost 50% shorter in the tissue adhesive cyanoacrylate group, a significant between-group difference (p = 0.003). Spontaneous bleeding in the donor area during the first 24 h was observed in 11.1% of the tissue adhesive cyanoacrylate group versus 88.9% of the suture group-a significant difference. No significant between-group difference was observed in postoperative pain, inflammation, or degree of healing over time. There were no cases of hyperesthesia or wound necrosis. Utilization of tissue adhesive cyanoacrylate rather than suture in palatal de-epithelialized gingival graft harvesting reduces postoperative bleeding during the first 24 h, as well as the operative time.

Unsupervised domain adaptation for the segmentation of breast tissue in mammography images.

BACKGROUND AND OBJECTIVE: Breast density refers to the proportion of glandular and fatty tissue in the breast and is recognized as a useful factor assessing breast cancer risk. Moreover, the segmentation of the high-density glandular tissue from mammograms can assist medical professionals visualizing and localizing areas that may require additional attention. Developing robust methods to segment breast tissues is challenging due to the variations in mammographic acquisition systems and protocols. Deep learning methods are effective in medical image segmentation but they often require large quantities of labelled data. Unsupervised domain adaptation is an area of research that employs unlabelled data to improve model performance on variations of samples derived from different sources. METHODS: First, a U-Net architecture was used to perform segmentation of the fatty and glandular tissues with labelled data from a single acquisition device. Then, adversarial-based unsupervised domain adaptation methods were used to incorporate single unlabelled target domains, consisting of images from a different machine, into the training. Finally, the domain adaptation model was extended to include multiple unlabelled target domains by combining a reconstruction task with adversarial training. RESULTS: The adversarial training was found to improve the generalization of the initial model on new domain data, demonstrating clearly improved segmentation of the breast tissues. For training with multiple unlabelled domains, combining a reconstruction task with adversarial training improved the stability of the training and yielded adequate segmentation results across all domains with a single model. CONCLUSIONS: Results demonstrated the potential for adversarial-based domain adaptation with U-Net architectures for segmentation of breast tissue in mammograms coming from several devices and demonstrated that domain-adapted models could achieve a similar agreement with manual segmentations. It has also been found that combining adversarial and reconstruction-based methods can provide a simple and effective solution for training with multiple unlabelled target domains.

Effect of Propolis Paste and Mouthwash Formulation on Healing after Teeth Extraction in Periodontal Disease.

This study investigated the antimicrobial effects of a mouthwash containing propolis and the effect of a propolis paste formulation on dental healing after teeth extraction in patients with periodontal disease. In the mouthwash experience, the population comprised 40 patients, which were divided as follows: the control mouthwash, 0.2% chlorhexidine (v/v) mouthwash, 2% (w/v) propolis mouthwash, and propolis + chlorhexidine mouthwash. The study of the propolis paste comprised a population of 60 patients with periodontal disease, and a total of 120 symmetric tooth extractions were performed. Propolis showed antimicrobial activity by itself, and especially with the chlorhexidine association. Three days after surgery in the teeth treated with control paste, only 13.4% had completely healed; however, with propolis paste, in 90% of the periodontal sockets, healing was complete. In addition, a reduction in Streptococci mutans and Lactobacilli cfu was observed with propolis, and especially with the association of chlorhexidine + propolis. Propolis mouthwash reduced bacterial proliferation, especially in association with chlorhexidine. Propolis paste is a viable alternative for socket healing after dental extraction. The knowledge gained from these findings will provide a foundation for similar propolis therapies in order to improve the healing process after dental surgery.

Comparison of Implant Stability between Regenerated and Non-Regenerated Bone. A Prospective Cohort Study.

Implant stability is one of the main indicators of successful osseointegration. Although it has been measured in numerous studies, there has been little research on implant stability in regenerated bone. The study compares primary and secondary stability between implants placed in regenerated versus native bone and evaluates the influence of bone quality on the results. Sixty implants were placed in 31 patients: 30 implants inserted in native bone (non-regenerated) after a healing period of at least 6 months post-exodontia and 30 inserted in regenerated bone at 6 months after grafting with xenograft. Resonance frequency analysis (RFA) was used to obtain implant stability quotient (ISQ) values at baseline (implant placement), 8 weeks, and 12 weeks. Statistically significant differences were found between implants placed in regenerated bone and those placed in native bone at all measurement time points (p < 0.05). ISQ values were significantly influenced by bone quality at baseline (p < 0.05) but not at 8 or 12 weeks. Greater stability was obtained in implants placed in native bone; however, those placed in regenerated bone showed adequate primary and secondary stability for prosthetic loading. Bone quality influences the primary but not secondary stability of the implants in both native and regenerated bone.

Composición corporal, metabolismo mineral y función endocrina del tejido adiposo: influencia de un suplemento nutricional de propóleo / Body composition, mineral metabolism, and endocrine function of adipose tissue: influence of a nutritional supplement of propolis

Introducción: el propóleo y sus componentes influyen en el metabolismo lipídico; sin embargo, se desconoce su efecto sobre la composición corporal y el metabolismo mineral. Objetivos: determinar el efecto de la suplementación de la dieta con propóleo natural sobre la composición corporal, el metabolismo basal y mineral, y la función endocrina del tejido adiposo. Material y métodos: veinte ratas albinas Wistar macho (8 semanas) se dividieron en dos grupos de 10 animales cada uno. Las ratas fueron alimentadas con dos tipos diferentes de dietas durante 90 días: una dieta estándar para el grupo de control (grupo C) y la misma dieta estándar + un 2 % de propóleo (grupo P). Se determinaron las hormonas tiroideas, la grelina, la leptina, la adiponectina y la insulina, los ácidos grasos no esterificados (AGNE) en el plasma, la composición corporal (masa magra, masa grasa y agua corporal) y el depósito de minerales en órganos diana (bazo, cerebro, corazón, pulmones, testículos, riñones y fémur). Resultados: los niveles plasmáticos de hormona estimulante del tiroides (TSH), triyodotironina (T3) y tiroxina (T4) no mostraron diferencias tras la ingesta del suplemento de propóleo, mientras que los de grelina y adiponectina disminuyeron (p < 0,01 y p < 0,05, respectivamente) y los de insulina (p < 0,01), leptina (p < 0,05) y AGNE (p < 0,05) aumentaron cuando la dieta se suplementó con propóleo al 2 %. Se redujeron el peso y la grasa corporal (p < 0,05), incrementándose la masa magra. Por último, el suplemento de propóleo mejoró el depósito de calcio en el bazo, los pulmones, los testículos y el fémur (p < 0,05). (AU)

Introduction: propolis and its components influence lipid metabolism; however, its effect on body composition and mineral metabolism remains unknown. Objectives: to determine the effect of natural propolis supplementation on body composition, mineral metabolism, and the endocrine function of adipose tissue. Material and methods: twenty albino male Wistar rats (8 weeks old) were divided into two groups of 10 animals each. The rats were fed two different types of diet for 90 days: a standard diet for the control group (group C) and the same standard diet + 2 % propolis (group P). Thyroid hormones, ghrelin, leptin, adiponectin and insulin, non-esterified fatty acids (NEFA) in plasma, body composition (lean mass, fat mass and body water), and mineral deposition in target organs (spleen, brain, heart, lungs, testicles, kidneys and femur) were assessed. Results: thyroid stimulating hormone (TSH), triiodothyronine (T3) and thyroxine (T4) did not show any differences after supplementation with propolis, while ghrelin and adiponectin decreased (p < 0.01 and p < 0.05, respectively) and insulin (p < 0.01), leptin (p < 0.05) and NEFA (p < 0.05) increased when 2 % propolis was supplied, while weight and body fat were reduced (p < 0.05) and lean mass increased. Lastly, the propolis supplement improves calcium deposition in the spleen, lungs, testes, and femur (p < 0.05). (AU)

[Body composition, mineral metabolism, and endocrine function of adipose tissue: influence of a nutritional supplement of propolis]. / Composición corporal, metabolismo mineral y función endocrina del tejido adiposo: influencia de un suplemento nutricional de propóleo.

INTRODUCTION: Introduction: propolis and its components influence lipid metabolism; however, its effect on body composition and mineral metabolism remains unknown. Objectives: to determine the effect of natural propolis supplementation on body composition, mineral metabolism, and the endocrine function of adipose tissue. Material and methods: twenty albino male Wistar rats (8 weeks old) were divided into two groups of 10 animals each. The rats were fed two different types of diet for 90 days: a standard diet for the control group (group C) and the same standard diet + 2 % propolis (group P). Thyroid hormones, ghrelin, leptin, adiponectin and insulin, non-esterified fatty acids (NEFA) in plasma, body composition (lean mass, fat mass and body water), and mineral deposition in target organs (spleen, brain, heart, lungs, testicles, kidneys and femur) were assessed. Results: thyroid stimulating hormone (TSH), triiodothyronine (T3) and thyroxine (T4) did not show any differences after supplementation with propolis, while ghrelin and adiponectin decreased (p < 0.01 and p < 0.05, respectively) and insulin (p < 0.01), leptin (p < 0.05) and NEFA (p < 0.05) increased when 2 % propolis was supplied, while weight and body fat were reduced (p < 0.05) and lean mass increased. Lastly, the propolis supplement improves calcium deposition in the spleen, lungs, testes, and femur (p < 0.05). Conclusion: propolis supplementation of the diet (2 %) causes a decrease in the secretion of ghrelin and adiponectin, increasing the release of non-esterified fatty acids and the rate of insulin secretion. In addition, propolis supplementation induces an improvement in calcium deposition in target organs without affecting the rest of minerals, which improves body composition by inducing a reduction in weight and visceral adipose tissue, and improvement in lean mass.

INTRODUCCIÓN: Introducción: el propóleo y sus componentes influyen en el metabolismo lipídico; sin embargo, se desconoce su efecto sobre la composición corporal y el metabolismo mineral. Objetivos: determinar el efecto de la suplementación de la dieta con propóleo natural sobre la composición corporal, el metabolismo basal y mineral, y la función endocrina del tejido adiposo. Material y métodos: veinte ratas albinas Wistar macho (8 semanas) se dividieron en dos grupos de 10 animales cada uno. Las ratas fueron alimentadas con dos tipos diferentes de dietas durante 90 días: una dieta estándar para el grupo de control (grupo C) y la misma dieta estándar + un 2 % de propóleo (grupo P). Se determinaron las hormonas tiroideas, la grelina, la leptina, la adiponectina y la insulina, los ácidos grasos no esterificados (AGNE) en el plasma, la composición corporal (masa magra, masa grasa y agua corporal) y el depósito de minerales en órganos diana (bazo, cerebro, corazón, pulmones, testículos, riñones y fémur). Resultados: los niveles plasmáticos de hormona estimulante del tiroides (TSH), triyodotironina (T3) y tiroxina (T4) no mostraron diferencias tras la ingesta del suplemento de propóleo, mientras que los de grelina y adiponectina disminuyeron (p < 0,01 y p < 0,05, respectivamente) y los de insulina (p < 0,01), leptina (p < 0,05) y AGNE (p < 0,05) aumentaron cuando la dieta se suplementó con propóleo al 2 %. Se redujeron el peso y la grasa corporal (p < 0,05), incrementándose la masa magra. Por último, el suplemento de propóleo mejoró el depósito de calcio en el bazo, los pulmones, los testículos y el fémur (p < 0,05). Conclusión: el suplemento de propóleo al 2 % de la dieta produjo una disminución de la secreción de grelina y adiponectina, incrementando la concentración de AGNE y aumentando la tasa de secreción de insulina. Además, el suplemento de propóleo indujo una mejora del depósito de calcio en los órganos diana sin afectar al resto de minerales, lo que en conjunto mejora la composición corporal al inducir una reducción del peso y del tejido adiposo visceral, mejorando la masa magra.

Allium-Based Phytobiotic Enhances Egg Production in Laying Hens through Microbial Composition Changes in Ileum and Cecum.

Phytobiotics (bioactive compounds extracted from plants) are one of the explored alternatives to antibiotics in poultry and livestock due to their antimicrobial activity and its positive effects on gut microbiota and productive properties. In this study, we supplemented a product based on garlic and onion compounds in the diet to laying hens at the beginning of their productive life (from 16 to 20 weeks post-hatching). The experimental group showed a significant increase in the number of eggs laid and in their size, produced in one month compared to the control. This increase in production was accompanied by microbiota changes in the ileum and cecum by means of high throughput sequencing analyses. These bacterial shifts in the ileum were mainly the result of compositional changes in the rare biosphere (unweighted UniFrac), while in the cecum, treatment affected both majority and minority bacterial groups (weighted and unweighted UniFrac). These changes in the microbiota suggest an improvement in food digestibility. The relative abundance of Lactococcus in the ileum and Lactobacillus in the cecum increased significantly in the experimental group. The relative abundance of these bacterial genera are known to have positive effects on the hosts. These results are very promising for the use of these compounds in poultry for short periods.

Mucinous ascites secondary to peritoneal pseudomyxoma of an uncertain origin.

Peritoneal pseudomyxoma (PP) is a rare malignant tumor, which is characterized by the presence of mucinous ascites. Abdominal ultrasound as the first diagnostic method of this entity allows the visualization of ascites on the one hand and its analysis through a diagnostic paracentesis on the other.

Identification of a Novel Variant in EARS2 Associated with a Severe Clinical Phenotype Expands the Clinical Spectrum of LTBL.

The EARS2 nuclear gene encodes mitochondrial glutamyl-tRNA synthetase, a member of the class I family of aminoacyl-tRNA synthetases (aaRSs) that plays a crucial role in mitochondrial protein biosynthesis by catalyzing the charging of glutamate to mitochondrial tRNA(Glu). Pathogenic EARS2 variants have been associated with a rare mitochondrial disorder known as leukoencephalopathy with thalamus and brainstem involvement and high lactate (LTBL). The targeted sequencing of 150 nuclear genes encoding respiratory chain complex subunits and proteins implicated in the oxidative phosphorylation (OXPHOS) function was performed. The oxygen consumption rate (OCR), and the extracellular acidification rate (ECAR), were measured. The enzymatic activities of Complexes I-V were analyzed spectrophotometrically. We describe a patient carrying two heterozygous EARS2 variants, c.376C>T (p.Gln126*) and c.670G>A (p.Gly224Ser), with infantile-onset disease and a severe clinical presentation. We demonstrate a clear defect in mitochondrial function in the patient's fibroblasts, suggesting the molecular mechanism underlying the pathogenicity of these EARS2 variants. Experimental validation using patient-derived fibroblasts allowed an accurate characterization of the disease-causing variants, and by comparing our patient's clinical presentation with that of previously reported cases, new clinical and radiological features of LTBL were identified, expanding the clinical spectrum of this disease.

Mutations Selected After Exposure to Bacteriocin Lcn972 Activate a Bce-Like Bacitracin Resistance Module in Lactococcus lactis.

Resistance against antimicrobial peptides (AMPs) is often mediated by detoxification modules that rely on sensing the AMP through a BceAB-like ATP-binding cassette (ABC) transporter that subsequently activates a cognate two-component system (TCS) to mount the cell response. Here, the Lactococcus lactis ABC transporter YsaDCB is shown to constitute, together with TCS-G, a detoxification module that protects L. lactis against bacitracin and the bacteriocin Lcn972, both AMPs that inhibit cell wall biosynthesis. Initially, increased expression of ysaDCB was detected by RT-qPCR in three L. lactis resistant to Lcn972, two of which were also resistant to bacitracin. These mutants shared, among others, single-point mutations in ysaB coding for the putative Bce-like permease. These results led us to investigate the function of YsaDCB ABC-transporter and study the impact of these mutations. Expression in trans of ysaDCB in L. lactis NZ9000, a strain that lacks a functional detoxification module, enhanced resistance to both AMPs, demonstrating its role as a resistance factor in L. lactis. When the three different ysaB alleles from the mutants were expressed, all of them outperformed the wild-type transporter in resistance against Lcn972 but not against bacitracin, suggesting a distinct mode of protection against each AMP. Moreover, P ysaD promoter fusions, designed to measure the activation of the detoxification module, revealed that the ysaB mutations unlock transcriptional control by TCS-G, resulting in constitutive expression of the ysaDCB operon. Finally, deletion of ysaD was also performed to get an insight into the function of this gene. ysaD encodes a secreted peptide and is part of the ysaDCB operon. YsaD appears to modulate signal relay between the ABC transporter and TCS-G, based on the different response of the P ysaD promoter fusions when it is not present. Altogether, the results underscore the unique features of this lactococcal detoxification module that warrant further research to advance in our overall understanding of these important resistance factors in bacteria.

[Positive effect of a propolis supplement on lipid profile, glycemia, and hepatic antioxidant status in an experimental animal model]. / Efecto positivo de un suplemento de propóleo sobre el perfil lipídico, la glucemia y el estado antioxidante hepático en un modelo animal experimental.

INTRODUCTION: Introduction: the use of natural nutritional supplements can be used as a coadjuvant therapy since they have several phytochemicals with potential antioxidant effects that could influence lipid and glycemic metabolism. Objectives: to determine the effect of natural propolis supplementation on lipid metabolism and liver antioxidant activity. Material and methods: 20 male Wistar albino rats (8 weeks) were divided into two groups of 10 animals each. Subsequently, they were fed two different types of diet for 90 days: a standard diet for the control group (diet C) and a standard diet + 2 % propolis (diet P). Hematological and biochemical parameters were assessed. The livers were extracted and washed with saline solution, and the cytosolic fractions were prepared fresh for additional analysis of antioxidant enzymes catalase, superoxide dismutase, glutathione peroxidase, and glutathione reductase. Results: after consuming the diet supplemented with propolis, we found a reduction in glucose (p < 0,01), total cholesterol (p < 0,001), GOT (p < 0,05) and GPT (p < 0,01), whereas the propolis supplement induced an increase in the hepatic activity of SOD (p < 0,001), CAT (p < 0,01) and GR (p < 0,05). Conclusion: the present study reveals that a dietary propolis supplement (2 %) can improve the lipid and glycemic profiles, also increasing antioxidant enzimes and reducing the release of liver transaminases.

INTRODUCCIÓN: Introducción: el uso de suplementos nutricionales naturales puede ser una terapia complementaria ya que tienen fitoquímicos con posibles efectos antioxidantes que pueden mejorar la regulación del metabolismo lipídico y glucémico. Objetivos: determinar el efecto de la suplementación con propóleo natural sobre el metabolismo lipídico y la actividad antioxidante hepática. Material y métodos: 20 ratas albinas Wistar macho (8 semanas) se dividieron en dos grupos de 10 animales cada uno y se sometieron a un período de 90 días en el que se alimentaron con dos tipos diferentes de dietas: estándar para el grupo de control (dieta C) y dieta estándar + 2 % de propóleo (dieta P). Se determinaron los parámetros hematológicos y bioquímicos. Se extrajeron los hígados, se lavaron con solución salina y las fracciones citosólicas se prepararon frescas para análisis adicionales de las enzimas antioxidantes catalasa, superóxido-dismutasa, glutatión-peroxidasa y glutatión-reductasa. Resultados: tras el consumo de la dieta con suplemento de propóleo, encontramos una reducción de la glucosa (p < 0,01), el colesterol total (p < 0,001), la GOT (p < 0,05) y la GPT (p < 0,01), mientras que dicha dieta indujo un incremento en la actividad hepática de SOD (p < 0,001), CAT (p < 0,01) y GR (p < 0,05). Conclusión: el presente estudio revela que el suplemento nutricional de propóleo al 2 % puede mejorar significativamente el perfil lipídic, el glucémico y las enzimas antioxidantes, y reducir la liberación de GOT y GPT hepáticas.