SARS-CoV-2 Specific Nanobodies Neutralize Different Variants of Concern and Reduce Virus Load in the Brain of h-ACE2 Transgenic Mice.

Since the beginning of the COVID-19 pandemic, there has been a significant need to develop antivirals and vaccines to combat the disease. In this work, we developed llama-derived nanobodies (Nbs) directed against the receptor binding domain (RBD) and other domains of the Spike (S) protein of SARS-CoV-2. Most of the Nbs with neutralizing properties were directed to RBD and were able to block S-2P/ACE2 interaction. Three neutralizing Nbs recognized the N-terminal domain (NTD) of the S-2P protein. Intranasal administration of Nbs induced protection ranging from 40% to 80% after challenge with the WA1/2020 strain in k18-hACE2 transgenic mice. Interestingly, protection was associated with a significant reduction in virus replication in nasal turbinates and a reduction in virus load in the brain. Employing pseudovirus neutralization assays, we identified Nbs with neutralizing capacity against the Alpha, Beta, Delta, and Omicron variants, including a Nb capable of neutralizing all variants tested. Furthermore, cocktails of different Nbs performed better than individual Nbs at neutralizing two Omicron variants (B.1.529 and BA.2). Altogether, the data suggest the potential of SARS-CoV-2 specific Nbs for intranasal treatment of COVID-19 encephalitis.

Valor Prognóstico da Escore PRECİSE DAPT em Resultados de Curto e Longo Prazo em Pacientes MINOCA com Síndrome Coronariana Aguda / Prognostic Value of PRECİSE DAPT Score on Short- and Long-Term Outcomes in MINOCA Patients with Acute Coronary Syndrome

Resumo Fundamento O infarto do miocárdio com artérias coronárias não obstrutivas (MINOCA) constitui um subconjunto significativo de infartos agudos do miocárdio (IAM) com marcadores prognósticos incertos. A avaliação precoce do risco é crucial para identificar pacientes MINOCA em risco de resultados adversos. Objetivos Este estudo teve como objetivo avaliar a capacidade preditiva do escore PRECISE-DAPT na avaliação do prognóstico de curto e longo prazo em pacientes MINOCA que apresentam infarto do miocárdio sem supradesnivelamento do segmento ST (IAMSSST) ou com supradesnivelamento do segmento ST (IAMCSST). Métodos Entre 741 pacientes MINOCA, o escore PRECISE-DAPT foi calculado para analisar sua associação com eventos cardiovasculares adversos maiores (MACE) intra-hospitalares e de acompanhamento. Os parâmetros que apresentaram significância nos grupos MACEM (+) foram submetidos à análise estatística: regressão logística univariada para eventos intra-hospitalares e regressão univariada de Cox para eventos de seguimento. Para significância estatística, foi adotado nível pré-definido de α = 0,05. Os parâmetros que demonstraram significância foram submetidos à regressão logística múltipla para eventos intra-hospitalares e à regressão multivariada de Cox para eventos de seguimento. Resultados Os MACE intra-hospitalares ocorreram em 4,1% dos pacientes, enquanto 58% apresentaram MACE no acompanhamento. Os níveis de hemoglobina e o escore PRECISE-DAPT foram identificados como parâmetros independentes para MACE intra-hospitalar. Além disso, a fração de ejeção (FE%) e o escore PRECISE-DAPT surgiram como preditores independentes de MACE no acompanhamento. Conclusões O estudo revelou que um escore PRECISE-DAPT mais alto foi significativamente associada a riscos aumentados de eventos cardiovasculares adversos maiores tanto intra-hospitalares quanto de longo prazo em pacientes MINOCA que apresentam síndrome coronariana aguda (SCA), ressaltando o potencial do escore na estratificação de risco para esta coorte de pacientes.

Abstract Background Myocardial infarction with non-obstructive coronary arteries (MINOCA) constitutes a significant subset of acute myocardial infarctions (AMI) with uncertain prognostic markers. Early risk assessment is crucial to identify MINOCA patients at risk of adverse outcomes. Objectives This study aimed to evaluate the predictive capacity of the PRECISE-DAPT score in assessing short- and long-term prognoses in MINOCA patients presenting with ST-segment elevation myocardial infarction (STEMI) or non-ST-elevation myocardial infarction (NSTEMI). Methods Among 741 MINOCA patients, the PRECISE-DAPT score was computed to analyze its association with in-hospital and follow-up major adverse cardiovascular events (MACE). Parameters showing significance in MACE (+) groups underwent statistical analysis: univariate logistic regression for in-hospital events and univariate Cox regression for follow-up events. For statistical significance, a predefined level of α = 0.05 was adopted. Parameters demonstrating significance proceeded to multiple logistic regression for in-hospital events and multivariate Cox regression for follow-up events. Results In-hospital MACE occurred in 4.1% of patients, while 58% experienced follow-up MACE. Hemoglobin levels and the PRECISE-DAPT Score were identified as independent parameters for in-hospital MACE. Furthermore, ejection fraction (EF%) and the PRECISE-DAPT Score emerged as independent predictors of follow-up MACE. Conclusions The study revealed that a higher PRECISE-DAPT score was significantly associated with increased risks of both in-hospital and long-term major adverse cardiovascular events in MINOCA patients presenting with acute coronary syndrome (ACS), underscoring the score's potential in risk stratification for this patient cohort.

Phytoremediation of toxic heavy metals in polluted soils and water of Dargai District Malakand Khyber Pakhtunkhwa, Pakistan / Fitorremediação de metais pesados ​​tóxicos em solos e água poluídos do distrito de Dargai Malakand Khyber Pakhtunkhwa, Paquistão

Abstract The contamination of natural resources with heavy metals released from steel mills is the primary cause of soil and water pollution in the Dargai Malakand, located on the northern side of Pakistan. Therefore, the present study was aimed to determine the level of heavy metals in soil and water samples of this area. The wild plant growing (nine native plants: Pteris vittata, Populus nigra, Eucalyptus camaldulensis, Persicaria maculosa, Arundo donax, Xanthium strumarium, Verbascum thapsus, Ricinus communis and Parthenium hysterophorus) there were then tested for their phytoremediation capabilities which is an environmentally friendly, generally utilized, and low-cost approach to eliminate heavy metals from polluted soils and water. Soil, water, and effluent samples were taken from the contaminated sites of seven steel mills in Dargai District Malakand and subjected to heavy metals analysis. Based on bioconcentration factor (BCF) and translocation factor (TF) calculated, The highest BCF for zinc was recorded for Pteris vittata roots (3.93), while the lowest value was observed for Verbascum thapsus leaves (0.306). Pteris vittata root showed the highest BCF for iron (1.618), while Ricinus communis leaves showed the lowest (0.023). The highest BCF value for chromium was highest for Populus nigra roots (0.717), while the lowest value was recorded for Persicaria maculosa leaves (0.031). For the selected metals; Fe, Zn and Cr the highest TF were recorded for Pteris vittata (0.988), Verbascum thapsus (0.944) and Xanthium strumairum (0.968) respectively. Therefore, it is recommended that these plants should be grown near to steel mills to reclaim heavy metals from industrial effluent, polluted soil as well as from polluted water.

A contaminação dos recursos naturais com metais pesados ​​liberados pelas siderúrgicas é a principal causa da poluição do solo e da água no Dargai Malakand, localizado no lado norte do Paquistão. Portanto, o presente estudo teve como objetivo determinar o teor de metais pesados ​​em amostras de solo e água desta área. O cultivo de plantas silvestres (nove plantas nativas: Pteris vittata, Populus nigra, Eucalyptus camaldulensis, Persicaria maculosa, Arundo donax, Xanthium strumarium, Verbascum thapsus, Ricinus communis e Parthenium hysterophorus) foi testado quanto às suas capacidades de fitorremediação, que é um meio ambiente, geralmente utilizado, e abordagem de baixo custo para eliminar metais pesados ​​de solos e águas poluídas. Amostras de solo, água e efluentes foram retiradas dos locais contaminados de sete siderúrgicas no distrito de Dargai, Malakand, e submetidas à análise de metais pesados. Com base no fator de bioconcentração (BCF) e fator de translocação (TF) calculados, o maior BCF para zinco foi registrado para as raízes de Pteris vittata (3,93), enquanto o menor valor foi observado para as folhas de Verbascum thapsus (0,306). A raiz de Pteris vittata apresentou o maior BCF para ferro (1,618), enquanto as folhas de Ricinus communis apresentaram o menor (0,023). O maior valor de BCF para cromo foi maior para raízes de Populus nigra (0,717), enquanto o menor valor foi registrado para folhas de Persicaria maculosa (0,031). Para os metais selecionados, Fe, Zn e Cr, os maiores TF foram registrados para Pteris vittata (0,988), Verbascum thapsus (0,944) e Xanthium strumairum (0,968), respectivamente. Portanto, recomenda-se que essas plantas sejam cultivadas próximas a siderúrgicas para recuperar metais pesados ​​de efluentes industriais, solo poluído e água poluída.

Determination of genetic diversity in Acacia modesta germplasm using SDS-PAGE / Determinação da diversidade genética em germoplasma de Acacia modesta usando SDS-PAGE

Biochemical markers such as protein are very important to determine genetic diversity among plant species in a given population which in turn is very important for breeders and farmers as they can then easily select the most appropriate variety to grow in a given locality. In this connection, the present study is aimed to evaluate genetic diversity in Acacia modesta germplasm through Sodium Dodecyl Sulphate Polyacrylamide Gel Electrophoresis (SDS-PAGE) technique. About 40 genotypes were subjected to SDS-PAGE analysis where a total of 12 polypeptide bands were observed in electrophoretogram. Out of which 16.67% were monomorphic while the remaining 83.33% were polymorphic. Variation found in B-2, 4, 5, 6, 7, 8, 9, 10, 11 and 12, were 20, 22.50, 32.50, 10, 2.50, 22.50, 15, 5, 2.50 and 75% respectively. Locus contribution toward genetic disagreement was 83.33%. Cluster analysis sorted all the genotypes into 9 clusters. The genotypes in one cluster were identical regarding protein profiling and showed less intra-specific genetic variation whereas differences were find from other genotypes.

Marcadores bioquímicos, como proteínas, são muito importantes para determinar a diversidade genética entre espécies de plantas em determinada população, o que, por sua vez, é muito importante para criadores e agricultores, pois eles podem selecionar facilmente a variedade mais adequada para crescer em certa localidade. Nesse sentido, o presente estudo tem como objetivo avaliar a diversidade genética em germoplasma de Acacia modesta por meio da técnica de Eletroforese em Gel de Poliacrilamida com Dodecil Sulfato de Sódio (SDS-PAGE). Cerca de 40 genótipos foram submetidos à análise SDS-PAGE, em que foi observado um total de 12 bandas polipeptídicas no eletroforetograma. Destes, 16,67% eram monomórficos, enquanto os 83,33% restantes eram polimórficos. As variações encontradas em B-2, 4, 5, 6, 7, 8, 9, 10, 11 e 12 foram de 20, 22,50, 32,50, 10, 2,50, 22,50, 15, 5, 2,50 e 75%, respectivamente. A contribuição do lócus para a discordância genética foi de 83,33%. A análise de agrupamento classificou todos os genótipos em 9 agrupamentos. Os genótipos em um cluster foram idênticos em relação ao perfil de proteínas e apresentaram menor variação genética intraespecífica, enquanto diferenças foram encontradas em outros genótipos.

Autocatalytic Mechanism in the Anaerobic Reduction of Metmyoglobin by Sulfide Species.

The mechanism of the metal centered reduction of metmyoglobin (MbFeIII) by sulfide species (H2S/HS-) under an argon atmosphere has been studied by a combination of spectroscopic, kinetic, and computational methods. Asymmetric S-shaped time-traces for the formation of MbFeII at varying ratios of excess sulfide were observed at pH 5.3 < pH < 8.0 and 25 °C, suggesting an autocatalytic reaction mechanism. An increased rate at more alkaline pHs points to HS- as relevant reactive species for the reduction. The formation of the sulfanyl radical (HS•) in the slow initial phase was assessed using the spin-trap phenyl N-tert-butyl nitrone. This radical initiates the formation of S-S reactive species as disulfanuidyl/ disulfanudi-idyl radical anions and disulfide (HSSH•-/HSS•2- and HSS-, respectively). The autocatalysis has been ascribed to HSS-, formed after HSSH•-/HSS•2- disproportionation, which behaves as a fast reductant toward the intermediate complex MbFeIII(HS-). We propose a reaction mechanism for the sulfide-mediated reduction of metmyoglobin where only ferric heme iron initiates the oxidation of sulfide species. Beside the chemical interest, this insight into the MbFeIII/sulfide reaction under an argon atmosphere is relevant for the interpretation of biochemical aspects of ectopic myoglobins found on hypoxic tissues toward reactive sulfur species.

Binding mechanism of disulfide species to ferric hemeproteins: The case of metmyoglobin.

The interactions of the heme iron of hemeproteins with sulfide and disulfide compounds are of potential interest as physiological signaling processes. While the interaction with hydrogen sulfide has been described computationally and experimentally, the reaction with disulfide, and specifically the molecular mechanism for ligand binding has not been studied in detail. In this work, we study the association process for disulfane and its conjugate base disulfanide at different pH conditions. Additionally, by means of advanced sampling techniques based on multiple steered molecular dynamics, we provide free energy profiles for ligand migration for both acid/base species, showing a similar behavior to the previously reported for the related H2S/HS¯ pair. Finally, we studied the ligand interchange reaction (H2O/H2S, HS¯ and H2O/HSSH, HSS¯) by means of hybrid quantum mechanics-molecular mechanics calculations. We show that the anionic species are able to displace more efficiently the H2O bound to the iron, and that the H-bond network in the distal cavity can help the neutral species to perform the reaction. Altogether, we provide a molecular explanation for the experimental information and show that the global association process depends on a fine balance between the migration towards the active site and the ligand interchange reaction.

Reduction of metmyoglobin by inorganic disulfide species.

The mechanism of the metal centered reduction of metmyoglobin (MbFeIII) by inorganic disulfide species has been studied by combined spectroscopic and kinetic analyses, under argon atmosphere. The process is kinetically characterized by biexponential time traces, for variable ratios of excess disulfide to protein, in the pH interval 6.6-8.0. Using UV-vis and resonance Raman spectroscopies, we observed that MbFeIII is converted into a low spin hexacoordinated ferric complex, tentatively assigned as MbFeIII(HSS-)/MbFeIII(SS2-), in an initial fast step. The complex is slowly converted into a pentacoordinated ferrous form, assigned as MbFeII according to the resonance Raman records. The reduction is a pH-dependent process, but independent of the initial disulfide concentration, suggesting the unimolecular decomposition of the intermediate complex following a reductive homolysis. We estimated the rate of the fast formation of the complex at pH 7.4 (kon = 3.7 × 103 M-1 s-1), and a pKa2 = 7.5 for the equilibrium MbFeIII(HSS-)/MbFeIII(SS2-). Also, we estimated the rate for the slow reduction at the same pH (kred = 10-2 s-1). A reaction mechanism compliant with the experimental results is proposed. This mechanistic study provides a differential kinetic signature for the reactions of disulfide compared to sulfide species on metmyoglobin, which may be considered in other hemeprotein systems.

Reproductive toxicity of maternal exposure to di(2-ethylhexyl)phthalate and butyl paraben (alone or in association) on both male and female Wistar offspring.

Parabens and phthalates are commonly found as contaminants in human fluids and are able to provoke reproductive toxicity, being considered endocrine disruptors. To evaluate the effects of phthalate and paraben, alone or in combination, on reproductive development of the offspring, female pregnant Wistar rats were allocated in six experimental groups: Three control groups (gavage [CG], subcutaneous [CS], and gavage + subcutaneous) received corn oil as vehicle, and the remaining groups were exposed to di(2-ethylhexyl)phthalate (DEHP) (500 mg/kg, gavage), butyl paraben (BP) (100 mg/kg, subcutaneously), or MIX (DEHP + BP), from Gestational Day 12 until Postnatal Day (PND) 21. The following parameters were assessed on the offspring: anogenital distance and weight at PND 1, nipple counting at PND 13, puberty onset, estrous cycle, weights of reproductive and detoxifying organs, histological evaluation of reproductive organs, and sperm evaluations (counts, morphology, and motility). Female pups from MIX group presented reduced body weight at PND 1, lower AGD, and decreased endometrium thickness. Male animals showed decreased body weight at PND 1 and lower number of Sertoli cells on DEHP and MIX groups, MIX group revealed increase of abnormal seminiferous tubules, DEHP animals presented delayed preputial separation and higher percentage of immotile sperms, and BP males presented diminished number of Leydig cells. In conclusion, the male offspring was more susceptible to DEHP toxicity; even when mixed to paraben, the main negative effects observed seem to be due to antiandrogenic phthalate action. On the other hand, DEHP seems to be necessary to improve the effects of BP on reducing estrogen-dependent and increasing androgen-dependent events.

Phonation Resistance Training Exercises (PhoRTE) With and Without Expiratory Muscle Strength Training (EMST) For Patients With Presbyphonia: A Noninferiority Randomized Clinical Trial.

OBJECTIVES: Presbyphonia negatively impacts quality of life in patients with age-related voice changes. A proof-of-concept study showed promise for high vocal intensity exercise to treat presbyphonia, which became the basis for a novel intervention for age-related voice changes known as Phonation Resistance Training Exercises (PhoRTE). Expiratory Muscle Strength Training (EMST) has also been proposed as an additional intervention to target and strengthen the aging respiratory system; however, EMST has undergone limited evaluation as an adjunct treatment for elderly patients undergoing voice therapy for presbyphonia. This study determined if the addition of EMST to PhoRTE voice therapy (PhoRTE + EMST) is at least as effective at voice improvement as PhoRTE alone. STUDY DESIGN: Prospective, randomized, controlled, single-blinded, non-inferiority. MATERIALS AND METHODS: Participants aged 55 years or older with a diagnosis of vocal fold atrophy were randomized to complete PhoRTE therapy or PhoRTE + EMST. The primary outcome was change in Voice Handicap Index-10 (VHI-10). Secondary outcomes included the Aging Voice Index, maximum expiratory pressure, and acoustic and aerodynamic measures of voice. Repeated measures linear mixed models were constructed to analyze outcomes at a significance level of α = 0.10. RESULTS: Twenty-six participants were recruited for the study, and 24 participants were randomized to either treatment arm. Sixteen participants completed the entire study. Both treatment arms showed statistically significant and clinically meaningful improvements in VHI-10 (PhoRTE mean [M] = -8.20, P < 0.001; PhoRTE + EMST M = -9.58, P < 0.001), and PhoRTE + EMST was noninferior to PhoRTE alone (P = 0.069). Both groups experienced a statistically significant pre-post treatment decrease (improvement) in AVI scores (PhoRTE M = -18.40, P = 0.004; PhoRTE + EMST M = -16.28, P = 0.005). PhoRTE+EMST had statistically significantly greater changes in maximum expiratory pressure compared to PhoRTE alone (PhoRTE M = 8.24 cm H2O, PhoRTE + EMST M = 32.63 cm H2O; P= 0.015). Some secondary acoustic and aerodynamic outcomes displayed trends toward improvement. CONCLUSION: This study demonstrates that voice therapy targeting high vocal intensity exercise (eg, PhoRTE) and EMST can play a role in improving voice outcomes for patients with presbyphonia.

Identification of novel variants in retinitis pigmentosa genes by whole-exome sequencing

SUMMARY OBJECTIVE: Retinitis pigmentosa is an inherited degenerative disorder causing severe retinal dystrophy and visual impairment, mainly with onset in the first or second decades. The next-generation sequencing has become an efficient tool to identify disease-causing mutations in retinitis pigmentosa. The aim of this retrospective study was to investigate novel gene variants and evaluate the utility of whole-exome sequencing in patients with retinitis pigmentosa. METHODS: The medical records of 20 patients with retinitis pigmentosa at Eskişehir City Hospital between September 2019 and February 2022 were analyzed retrospectively. Peripheral venous blood was obtained, followed by the extraction of genomic DNAs. The medical and ophthalmic histories were collected, and ophthalmological examinations were performed. Whole-exome sequencing was performed to determine the genetic etiology of the patients. RESULTS: The proportion of genetically solved cases was 75% (15/20) in the patients with retinitis pigmentosa. Molecular genetic testing identified 13 biallelic and 4 monoallelic mutations in known retinitis pigmentosa genes, including 11 novel variants. According to in silico prediction tools, nine variants were predicted as pathogenic or possibly pathogenic. We identified six previously reported mutations to be associated with retinitis pigmentosa. The age of onset of the patients ranged from 3 to 19, with a mean age of onset of 11.6. All patients had a loss of central vision. CONCLUSION: As the first study of the application of whole-exome sequencing among patients with retinitis pigmentosa in a Turkish cohort, our results may contribute to the characterization of the spectrum of variants related to retinitis pigmentosa in the Turkish population. Future population-based studies will enable us to reveal the detailed genetic epidemiology of retinitis pigmentosa.

Feral dogs in Chitral gol national park, Pakistan: a potential threat to the future of threatened Kashmir Markhor (Capra falconeri cashmiriensis) / Cães ferozes no parque nacional Chitral gol, Paquistão: uma ameaça potencial para o futuro do Markhor da Caxemira ameaçado (Capra falconeri cashmiriensis)

Feral dogs are well-organized hunters of ungulates in many parts of the world, causing great damage to wildlife populations and ultimately to the ecosystem. In Pakistan, the impacts of feral dogs on the wildlife have not been documented yet. In a period of fifteen years (2006-2020), feral dogs have killed hundreds of threatened markhor in Chitral gol national park (CGNP), Pakistan. Despite direct predation other impacts including disturbance and competition with other natural predators, could compromise conservation and management efforts. The population of feral dogs seems to have been increased with the increase of dumping sites by communities. Our findings suggest that there are pressing needs of controlling the feral dogs population and eradicating them from the core zone of CGNP and surrounding buffer communities. Conventional culling of dogs should be coupled with modern techniques like castration and sterilization. Communities should be educated regarding the clean environment, proper disposal of home wastes and, biodiversity conservation.

Os cães ferozes são caçadores bem organizados de ungulados em muitas partes do mundo, causando grandes danos à população de animais selvagens e, em última instância, ao ecossistema. No Paquistão, os impactos dos cães selvagens na vida selvagem ainda não foram documentados. Em um período de 15 anos (2006-2020), cães selvagens mataram centenas de markhor ameaçados no parque nacional Chitral gol (CGNP), Paquistão. Apesar da predação direta, outros impactos, incluindo distúrbios e competições com outros predadores naturais podem comprometer os esforços de conservação e manejo. A população de cães selvagens parece ter aumentado com o aumento dos locais de despejo pelas comunidades. Nossas descobertas sugerem que há necessidades urgentes para controlar a população de cães selvagens e erradicá-los da zona central do CGNP e das comunidades-tampão vizinhas. O abate convencional de cães deve ser combinado com técnicas modernas como castração e esterilização. As comunidades devem ser educadas sobre o meio ambiente limpo, o descarte adequado de resíduos domésticos e a conservação da biodiversidade.

Molecular detection and prevalence of Rotavirus with acute gastroenteritis among the children of rural and urban areas / Detecção molecular e prevalência de rotavírus com gastroenterite aguda entre as crianças de áreas rurais e urbanas

Rotavirus is the main infective agent of acute gastroenteritis (AGE) in children under the age of five years and causing significant morbidity as well as mortality throughout the world. The study was carried out to detect the prevalence rate, genotypes strain and risk factors of Rotavirus among the children of rural and urban areas of district Bannu Khyber Pakhtunkhwa Pakistan. A total of 180 stool samples were collected from children under the age of 5 years from two major hospitals of Bannu from January to December (2015). The samples were analyzed by Reverse-transcriptase Polymerase Chain Reaction (RT-PCR) for the detection of Rotavirus, positive samples were further processed for genotyping (G and P type) through specific PCR. Of the total, 41 (23%) samples were positive for Rotavirus. The most prevalent G genotypes found were: G3, G8, G9 (each 29%), followed by G10 (15%), and G11 (10%). Whereas the prevalent P genotypes were: P-8 (25%), P-4 and P-10 (each 20%), P-9 (15%), followed by P-6 and P-11 (each 10%). Moreover, Rotavirus infection was more prevalent in summer (23.73%) and winter (22.7%) than spring (20%) and autumn (21.4%). Rotavirus infection exhibited high frequency in June (14%), October (8%) and November (6%). It is concluded that Rotavirus is more prevalent in children and various genotypes (G and P) of Rotavirus are present in the study area. Lack of studies, awareness and rarer testing of Rotavirus are the principal reasons of virus prevalence in district Bannu, Pakistan.

O rotavírus é o principal agente infeccioso da gastroenterite aguda (AGE) em crianças menores de 5 anos e causa de morbidade e mortalidade significativas em todo o mundo. O estudo foi realizado para detectar a taxa de prevalência, cepa de genótipos e fatores de risco de rotavírus entre as crianças de áreas rurais e urbanas do distrito de Bannu Khyber Pakhtunkhwa, Paquistão. Um total de 180 amostras de fezes foi coletada de crianças menores de 5 anos de dois grandes hospitais de Bannu de janeiro a dezembro (2015). As amostras foram analisadas por reação em cadeia da polimerase transcriptase reversa (RT-PCR) para detecção de rotavírus; as amostras positivas foram posteriormente processadas para genotipagem (tipo G e P) através de PCR específica. Do total, 41 (23%) amostras foram positivas para rotavírus. Os genótipos G mais prevalentes encontrados foram: G3, G8, G9 (cada 29%), seguidos de G10 (15%) e G11 (10%). Considerando que os genótipos P prevalentes foram: P-8 (25%), P-4 e P-10 (cada 20%), P-9 (15%), seguido por P-6 e P-11 (cada 10%). Além disso, a infecção por rotavírus foi mais prevalente no verão (23,73%) e inverno (22,7%) do que na primavera (20%) e no outono (21,4%). A infecção por rotavírus apresentou alta frequência em junho (14%), outubro (8%) e novembro (6%). Conclui-se que o rotavírus é mais prevalente em crianças e vários genótipos (G e P) do rotavírus estão presentes na área de estudo. A falta de estudos, conhecimento e testes mais raros de rotavírus são as principais razões da prevalência do vírus no distrito de Bannu, Paquistão.

Feral dogs in Chitral gol national park, Pakistan: a potential threat to the future of threatened Kashmir Markhor (Capra falconeri cashmiriensis)

Abstract Feral dogs are well-organized hunters of ungulates in many parts of the world, causing great damage to wildlife populations and ultimately to the ecosystem. In Pakistan, the impacts of feral dogs on the wildlife have not been documented yet. In a period of fifteen years (2006-2020), feral dogs have killed hundreds of threatened markhor in Chitral gol national park (CGNP), Pakistan. Despite direct predation other impacts including disturbance and competition with other natural predators, could compromise conservation and management efforts. The population of feral dogs seems to have been increased with the increase of dumping sites by communities. Our findings suggest that there are pressing needs of controlling the feral dogs population and eradicating them from the core zone of CGNP and surrounding buffer communities. Conventional culling of dogs should be coupled with modern techniques like castration and sterilization. Communities should be educated regarding the clean environment, proper disposal of home wastes and, biodiversity conservation.

Resumo Os cães ferozes são caçadores bem organizados de ungulados em muitas partes do mundo, causando grandes danos à população de animais selvagens e, em última instância, ao ecossistema. No Paquistão, os impactos dos cães selvagens na vida selvagem ainda não foram documentados. Em um período de 15 anos (2006-2020), cães selvagens mataram centenas de markhor ameaçados no parque nacional Chitral gol (CGNP), Paquistão. Apesar da predação direta, outros impactos, incluindo distúrbios e competições com outros predadores naturais podem comprometer os esforços de conservação e manejo. A população de cães selvagens parece ter aumentado com o aumento dos locais de despejo pelas comunidades. Nossas descobertas sugerem que há necessidades urgentes para controlar a população de cães selvagens e erradicá-los da zona central do CGNP e das comunidades-tampão vizinhas. O abate convencional de cães deve ser combinado com técnicas modernas como castração e esterilização. As comunidades devem ser educadas sobre o meio ambiente limpo, o descarte adequado de resíduos domésticos e a conservação da biodiversidade.

Molecular detection and prevalence of Rotavirus with acute gastroenteritis among the children of rural and urban areas

Abstract Rotavirus is the main infective agent of acute gastroenteritis (AGE) in children under the age of five years and causing significant morbidity as well as mortality throughout the world. The study was carried out to detect the prevalence rate, genotypes strain and risk factors of Rotavirus among the children of rural and urban areas of district Bannu Khyber Pakhtunkhwa Pakistan. A total of 180 stool samples were collected from children under the age of 5 years from two major hospitals of Bannu from January to December (2015). The samples were analyzed by Reverse-transcriptase Polymerase Chain Reaction (RT-PCR) for the detection of Rotavirus, positive samples were further processed for genotyping (G and P type) through specific PCR. Of the total, 41 (23%) samples were positive for Rotavirus. The most prevalent G genotypes found were: G3, G8, G9 (each 29%), followed by G10 (15%), and G11 (10%). Whereas the prevalent P genotypes were: P-8 (25%), P-4 and P-10 (each 20%), P-9 (15%), followed by P-6 and P-11 (each 10%). Moreover, Rotavirus infection was more prevalent in summer (23.73%) and winter (22.7%) than spring (20%) and autumn (21.4%). Rotavirus infection exhibited high frequency in June (14%), October (8%) and November (6%). It is concluded that Rotavirus is more prevalent in children and various genotypes (G and P) of Rotavirus are present in the study area. Lack of studies, awareness and rarer testing of Rotavirus are the principal reasons of virus prevalence in district Bannu, Pakistan.

Resumo O rotavírus é o principal agente infeccioso da gastroenterite aguda (AGE) em crianças menores de 5 anos e causa de morbidade e mortalidade significativas em todo o mundo. O estudo foi realizado para detectar a taxa de prevalência, cepa de genótipos e fatores de risco de rotavírus entre as crianças de áreas rurais e urbanas do distrito de Bannu Khyber Pakhtunkhwa, Paquistão. Um total de 180 amostras de fezes foi coletada de crianças menores de 5 anos de dois grandes hospitais de Bannu de janeiro a dezembro (2015). As amostras foram analisadas por reação em cadeia da polimerase transcriptase reversa (RT-PCR) para detecção de rotavírus; as amostras positivas foram posteriormente processadas para genotipagem (tipo G e P) através de PCR específica. Do total, 41 (23%) amostras foram positivas para rotavírus. Os genótipos G mais prevalentes encontrados foram: G3, G8, G9 (cada 29%), seguidos de G10 (15%) e G11 (10%). Considerando que os genótipos P prevalentes foram: P-8 (25%), P-4 e P-10 (cada 20%), P-9 (15%), seguido por P-6 e P-11 (cada 10%). Além disso, a infecção por rotavírus foi mais prevalente no verão (23,73%) e inverno (22,7%) do que na primavera (20%) e no outono (21,4%). A infecção por rotavírus apresentou alta frequência em junho (14%), outubro (8%) e novembro (6%). Conclui-se que o rotavírus é mais prevalente em crianças e vários genótipos (G e P) do rotavírus estão presentes na área de estudo. A falta de estudos, conhecimento e testes mais raros de rotavírus são as principais razões da prevalência do vírus no distrito de Bannu, Paquistão.

Feral dogs in Chitral gol national park, Pakistan: a potential threat to the future of threatened Kashmir Markhor (Capra falconeri cashmiriensis) / Cães ferozes no parque nacional Chitral gol, Paquistão: uma ameaça potencial para o futuro do Markhor da Caxemira ameaçado (Capra falconeri cashmiriensis)

Abstract Feral dogs are well-organized hunters of ungulates in many parts of the world, causing great damage to wildlife populations and ultimately to the ecosystem. In Pakistan, the impacts of feral dogs on the wildlife have not been documented yet. In a period of fifteen years (2006-2020), feral dogs have killed hundreds of threatened markhor in Chitral gol national park (CGNP), Pakistan. Despite direct predation other impacts including disturbance and competition with other natural predators, could compromise conservation and management efforts. The population of feral dogs seems to have been increased with the increase of dumping sites by communities. Our findings suggest that there are pressing needs of controlling the feral dogs population and eradicating them from the core zone of CGNP and surrounding buffer communities. Conventional culling of dogs should be coupled with modern techniques like castration and sterilization. Communities should be educated regarding the clean environment, proper disposal of home wastes and, biodiversity conservation.

Resumo Os cães ferozes são caçadores bem organizados de ungulados em muitas partes do mundo, causando grandes danos à população de animais selvagens e, em última instância, ao ecossistema. No Paquistão, os impactos dos cães selvagens na vida selvagem ainda não foram documentados. Em um período de 15 anos (2006-2020), cães selvagens mataram centenas de markhor ameaçados no parque nacional Chitral gol (CGNP), Paquistão. Apesar da predação direta, outros impactos, incluindo distúrbios e competições com outros predadores naturais podem comprometer os esforços de conservação e manejo. A população de cães selvagens parece ter aumentado com o aumento dos locais de despejo pelas comunidades. Nossas descobertas sugerem que há necessidades urgentes para controlar a população de cães selvagens e erradicá-los da zona central do CGNP e das comunidades-tampão vizinhas. O abate convencional de cães deve ser combinado com técnicas modernas como castração e esterilização. As comunidades devem ser educadas sobre o meio ambiente limpo, o descarte adequado de resíduos domésticos e a conservação da biodiversidade.

Importance of targeted next-generation sequencing in pediatric patients with developmental epileptic encephalopathy

SUMMARY OBJECTIVE: Childhood epilepsy is a common neurological disorder with a prevalence of 300-600 cases per 100,000 people. It is associated with refractory epilepsies, global developmental delay, and epileptic encephalopathies, causing epileptic syndromes characterized by cognitive and behavioral disorders. METHODS: In this retrospective cohort study, patients with refractory epilepsy and global developmental delay, defined as epileptic encephalopathy, who applied to the Aydın 7Maternity and Children's Hospital Genetic Diagnosis Center and were followed in the pediatric neurology clinic of our hospital, between July 2018 and July 2021, were included. RESULTS: Targeted next-generation sequencing molecular genetics results were reviewed, and 3 ALDH7A1, 1 AARS, 3 CACNA1A, 1 CTNNB1, 1 DCX, 2 DBH, 2 DOCK7, 1 FOLR1, 2 GABRB3, 2 GCH1, 1 VGRIN2B, 1 GUF1, 3 KCNQ2, 2 KCNT1, 1 NECAP1, 1 PCDH19, 1 PNPO, 1 SCN8A, 1 SCN9A, 4 SCN1A, 2 SLC25A22, 1 SLC2A1, 2 SPTAN1, 2 SZT2, 4 TBC1D24, 2 TH, and 1 PCDH19 (X chromosome) mutations were detected in three of the patients using the next-generation sequencing method. CONCLUSION: Although the development of gene panels aids in diagnosis, there are still unidentified disorders in this illness category, which is highly variable in genotype and phenotype. Understanding the genetic etiology is vital for genetic counseling and, maybe, the future development of remedies for the etiology.