The coronavirus disease (COVID-19) pandemic, caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), has had a profound impact on global health and socio-economic conditions. To date, various vaccines have been administered worldwide in an effort to curb the spread of the virus. Despite vaccination efforts, there have been complications. Existing antiviral drugs have shown limited effectiveness, prompting the use of computational methods to understand the dynamics of the virus and develop suitable treatments. The current study focuses on using biocompatible para-sulfonato-calix[4]arenes to dock against two key proteins of SARS-CoV-2, namely ribonucleic acid (RNA)-dependent RNA polymerase, and helicase. Docking results indicate a strong binding affinity of these compounds to the target proteins, with higher scores compared to commonly used medications. Molecular dynamics (MD) simulation validates the docking results, showing stable protein-ligand complexes over time. The compounds are also screened for absorption, distribution, metabolism, and excretion properties and toxicity, suggesting their potential as lead candidates for inhibiting the virus’s key proteins. However, further in vivo and in vitro studies are recommended to confirm these findings. COVID19 COVID 19 (COVID-19 pandemic SARSCoV2, SARSCoV2 SARSCoV SARS CoV , (SARS-CoV-2) socioeconomic socio economic conditions date efforts complications effectiveness treatments parasulfonatocalix4arenes parasulfonatocalixarenes para sulfonato calix 4 arenes 2, SARS-CoV-2 RNAdependent dependent polymerase helicase medications MD (MD proteinligand protein ligand time absorption distribution metabolism toxicity viruss s However findings COVID1 1 (COVID-1 (SARS-CoV-2 parasulfonatocalix SARS-CoV- (COVID- (SARS-CoV- SARS-CoV (COVID (SARS-CoV

Introdução: Gliomas são os tumores primários mais comuns do sistema nervoso central e a maioria é formada por glioblastomas, com graus elevados de proliferação e invasão, com heterogeneidade biológica e pouca resposta aos tratamentos atuais. Imunoterapia e geneterapia não têm mostrado eficácia e, por isso, é necessário buscar conhecimento de novas moléculas e genes envolvidos na sua carcinogênese. Objetivos: Avaliar se o receptor do ácido gama-aminobutírico alfa 5 (GABRA5) é potencial alvo terapêutico ou biomarcador para gliomas de diferentes subgrupos, e a correlação entre os níveis de ácido ribonucleico mensageiro (mRNA) de GABRA5 com a sobrevida global. Métodos: Revisão integrativa feita em plataformas virtuais, em português e inglês, realizada por descritores relacionados ao tema, descritos no DeCS como “glioma, sistema nervoso central, glioblastoma” com busca AND ou OR, considerando-se inicialmente o título e/ou resumo. Após, naqueles que tinham maior relação ao tema, foi realizada a leitura na íntegra dos textos. Resultados: Foram incluídos 36 artigos. Conclusão: Os níveis de mRNA de GABRA5 em amostras de gliomas de todos os subgrupos histológicos são menores, em comparação ao controle. Não foi observada correlação entre os níveis de mRNA de GABRA5 com a sobrevida global dos subgrupos avaliados.

Introduction: Gliomas are the most common primary tumors of the central nervous system. The majority are formed by glioblastomas, with high degrees of proliferation and invasion, with biological heterogeneity and little response to current treatments. Immunotherapy and genetherapy have not shown efficacy and, therefore, it is necessary to seek knowledge of new molecules and genes involved in their carcinogenesis. Objectives: To evaluate whether the alpha 5 gamma-aminobutyric acid receptor (GABRA5) is a potential therapeutic target or biomarker for gliomas of different subgroups, and the correlation between GABRA5 messenger ribonucleic acid (mRNA) levels and overall survival. Methods: Integrative review carried out on virtual platforms in Portuguese and English, carried out using descriptors related to the topic, described in DeCS as “glioma, central nervous system, glioblastoma, GABA, GABRA5” with AND or OR search, initially considering the title and/or summary. Afterwards, in those that were most related to the topic, the texts were read in full. Results: 36 articles were included. Conclusion: GABRA5 mRNA levels in glioma samples from all histological subgroups are lower compared to control. No correlation was observed between GABRA5 mRNA levels and overall survival in the evaluated subgroups.

Abstract Background Cutaneous squamous cell carcinoma (cSCC) develops from epithelial keratinocytes by dysregulation of self-renewal and differentiation. Recent studies have found that the size and number of cSCC tumors gradually decrease or even disappear after HPV vaccination. However, the role of the HPV vaccine in the cSCC mechanism is poorly understood. Objective The aim of this study is to investigate the effect and mechanism of the HPV vaccine in cSCC. Methods Immunofluorescence was used to study the immune infiltrating cells in the tumor tissues of patients with cSCC. The effects of the HPV vaccine on cSCC cells and tissues were studied by Cell Culture, Real-time PCR, Western Blot, Cytotoxicity Assay, Enzyme-linked Immunosorbent Assay, m6A Blotting, CCK-8 Assay, m6A Ribonucleic acid Methylation Quantification and tumor transplantation. Results The HPV vaccine enhanced the toxic effect of CD8+T cells on cSCC cells and promoted the secretion of multiple cytokines by CD8+T cells. In addition, HPV vaccines can increase tumor sensitivity to anti-PD-1 therapy by downregulating METTL3 in tumor tissue, with the combination of HPV vaccine and PD-1 monoclonal antibodies producing enhanced immune cell infiltration compared to PD-1 blockade alone. Study limitations It is important to note the limitations of this study, including the small sample size, the construction of the mouse model, and the choice of HPV vaccine and PD-1 monoclonal antibody, which may limit the generalization of our findings to a wider population. Conclusions It is hoped that this research will contribute to a deeper understanding of the role of the HPV vaccine in the treatment of cSCC. HPV vaccine is expected to become an important approach to alleviate the development of cSCC. (cSCC selfrenewal self renewal differentiation vaccination However understood Culture Realtime Real time PCR Blot Assay Enzymelinked Enzyme linked mA m A Blotting CCK8 CCK 8 CCK- transplantation CD8T CDT CD8 T CD addition antiPD1 antiPD anti PD 1 anti-PD- METTL tissue PD1 PD- alone model antibody population anti-PD

SUMMARY OBJECTIVE: In the hepatitis C virus (HCV) diagnostic algorithm, an anti-HCV screening test is recommended first. In countries with low HCV prevalence, anti-HCV testing can often give false-positive results. This may lead to unnecessary retesting, increased costs, and psychological stress for patients. METHODS: In this study, the most appropriate S/Co (signal-cutoff) value to predict HCV viremia in anti-HCV test(+) individuals was determined, and the effect of genotype differences was evaluated. Of the 96,515 anti-HCV tests performed between 2020 and 2023, 934 were reactive. A total of 332 retests and 65 patients without HCV-ribonucleic acid (RNA) analysis were excluded. Demographic data were calculated for 537 patients, and 130 patients were included in the study. RESULTS: The average age of 537 patients was 55±18 years, and 57.1% were women. The anti-HCV positivity rate was 0.62% (602/96,515), and the actual anti-HCV positivity rate was 0.13% (130/96,515). Anti-HCV levels were higher in HCV-RNA(+) patients than in HCV-RNA-negative individuals (p<0.0001) (Table 1). Receiver operating characteristic curve analysis identified the optimal S/Co value to be 10.86 to identify true positive cases. Sensitivity was 96.1%, specificity was 61.2%, positive predictive value (PPV) was 44.2%, and negative predictive value (NPV) was 98% (Figure 2). A total of 107 (82.3%) of the patients were identified as GT1, and the most common subtype was GT1b (n=100). CONCLUSION: If anti-HCV S/Co is ≥10.86, direct HCV RNA testing may be recommended; However, the possibility of false positivity should be considered in patients with a S/Co value below 10.86. OBJECTIVE (HCV algorithm antiHCV anti first prevalence falsepositive results retesting costs METHODS study SCo S Co signalcutoff signal cutoff (signal-cutoff test+ + test(+ determined evaluated 96515 96 515 96,51 202 2023 93 reactive 33 6 HCVribonucleic ribonucleic (RNA excluded 53 13 RESULTS 5518 55 18 55±1 years 571 57 1 57.1 women 062 0 62 0.62 602/96,515, 60296515 602/96,515 , 602 (602/96,515) 013 0.13 130/96,515. 13096515 130/96,515 . (130/96,515) AntiHCV Anti HCVRNA+ HCVRNA HCV-RNA(+ HCVRNAnegative p<0.0001 p00001 p 0001 (p<0.0001 Table 1. 1) 1086 10 86 10.8 cases 961 96.1% 612 61 2 61.2% PPV (PPV 442 44 44.2% NPV (NPV 98 Figure 2. 2) 82.3% 823 82 3 (82.3% GT1 GT GTb b n=100. n100 n n=100 100 (n=100) CONCLUSION ≥10.86 However test( 9651 9 51 96,5 20 5 551 55± 57. 06 0.6 6029651 602/96,51 60 (602/96,515 01 0.1 1309651 130/96,51 (130/96,515 HCV-RNA( p<0.000 p0000 000 (p<0.000 108 8 10. 96.1 61.2 4 44.2 82.3 (82.3 n10 n=10 (n=100 ≥10.8 965 96, 0. 602965 602/96,5 (602/96,51 130965 130/96,5 (130/96,51 HCV-RNA p<0.00 p000 00 (p<0.00 96. 61. 44. 82. (82. n1 n=1 (n=10 ≥10. 60296 602/96, (602/96,5 13096 130/96, (130/96,5 p<0.0 p00 (p<0.0 (82 n= (n=1 ≥10 6029 602/96 (602/96, 1309 130/96 (130/96, p<0. p0 (p<0. (8 (n= ≥1 602/9 (602/96 130/9 (130/96 p<0 (p<0 ( (n ≥ 602/ (602/9 130/ (130/9 p< (p< (602/ (130/ (p (602 (130 (60 (13 (6 (1

Abstract Introduction Finding biomarkers for highly lethal cancers is a priority. Objective The current study was designed to understand the clinical significance of vascular endothelial growth factor (VEGF), latent membrane protein 1 (LMP1), and tumor necrosis factor-α (TNF-α) expression as the biomarkers, and evaluate their correlation with each other, in nasopharyngeal carcinoma (NPC) in the province of Guilan, North of Iran. Methods Gene expression was evaluated in 25 formalin-fixed paraffin-embedded (FFPE) blocks from cases of confirmed NPC and 20 FFPE samples of non-NPC by quantifying messenger ribonucleic acid (mRNA) and protein levels, using real-time polymerase chain reaction (PCR) and immunohistochemistry (IHC) methods, respectively. Furthermore, the correlations among the protein levels of different genes, along with the patients’ demographic characteristics were assessed. Results Our findings on mRNA and protein levels demonstrated that the expression of the LMP1 gene in the NPC group was significantly elevated compared with that of the non-NPC group. In addition, the protein levels in the NPC group indicated a positive and significant correlation between LMP1 and VEGF expression. It was noted that both protein and mRNA levels showed no significant differences in the expression of TNF-α and VEGF genes between the NPC and control groups. Furthermore, there was no significant relationship between the expression of these proteins and the demographic characteristics of NPC patients. Conclusion Overall, a significant increase in LMP1 expression was observed in NPC patients, which may serve as a diagnostic biomarker for NPC. Also, LMP1 might be involved in NPC progression by inducing VEGF gene expression. priority VEGF, , (VEGF) LMP1, LMP (LMP1) factorα α TNFα TNF (TNF-α other (NPC Guilan Iran 2 formalinfixed formalin fixed paraffinembedded paraffin embedded (FFPE nonNPC non (mRNA realtime real time PCR (PCR IHC (IHC methods respectively Furthermore patients assessed addition groups Overall Also (VEGF (LMP1 (LMP

The origin of humans has been a subject of intense debate for many centuries. The two main approaches to this subject are materialistic and metaphysical. The materialist view suggests that humans evolved from non-human primates through the process of natural selection, while the metaphysical view suggests that humans were created by a divine entity or a supernatural force. According to materialists, their view is based on the scientific theory of evolution, which suggests that all living organisms, including humans, descend from a common ancestor. According to this theory, the process of natural selection drives the development of species over time and those individuals, with traits best suited to their environment, are more likely to survive and reproduce. Recent studies have provided convincing evidence for the descent with modification of humans from non-human primates, including genetic similarities, fossil evidence and anatomical comparisons. In contrast, the metaphysical view states that humans were created by a divine entity or a supernatural force. This view is often associated with religious beliefs and (according to its opponents) is not based on empirical evidence. For the origin of life there is a clear twofold requirement: firstly, a blueprint (plan) that determines how the organism will function (such as deoxyribonucleic acid (DNA) or ribonucleic acid (RNA)) and secondly a mechanism with a metabolic system to interpret the blueprint and functionally organise the proteins as described by the DNA or RNA. This is a classic "chicken-egg" situation, because for life to arise, a blueprint is needed according to which the first organism will be produced, while at the same time an organism is needed to produce the first blueprint. All living matter (biomolecules such as nucleic acids, proteins and lipids) consist of the same macro-nutrients, namely, carbon, hydrogen, nitrogen, oxygen, phosphate, and sulphur, of which carbon and water are the most essential. These shared building materials are signs of a common origin, which is confirmed in genetics by aspects such as shared cell design, genetic code, protein systems, as well as DNA, mRNA (messenger ribonucleic acid), and protein functions. The interconnectedness is especially clear where variations occur, such as changes through mutations which accumulate in the genome, especially in pseudogenes, where shared mutations then indicate shared ancestry; where retroviruses leave a copy (deactivated so-called "junk" genes) in the DNA of infected organisms, from which it is transmitted to the offspring to indicate relationships; and the sharing of parallel genes by some metazoa that indicate relationships. From a total of 6.1 billion prokaryotic protein-coding genes, 355 protein clusters have been identified that are shared by all prokaryotes. According to this, LUCA (last universal common ancestor) could successfully inhabit anaerobic hydrothermal vents on the sea floor in a H2, CO2 and iron-rich environment. This finding is consistent with the oldest evidence of life, namely microbial mats of bacteria and archaea that were the dominant life form during the earliest times of life on earth. Development and specialisation occurred during times of accelerated biodiversity, of which the Cambrian "explosion" was exceptional in scope and depth. DNA comparisons show that the phyla of the animal kingdom split into three groups, about 50 million years before the Cambrian period. The presence of complex DNA in all three groups that shared many characteristics, makes a complex ancestor a necessity. This indicates that the Cambrian biodiversification was driven by selection from the loaded genetic potential space of a single, shared ancestor. As a species, human DNA shows a much lower diversity than the genetic theory predicts, mainly due to its recency as well as the occurrence of a "population bottleneck", as indicated by the presence of a fusion chromosome. This fusion chromosome that once formed in a specific person, has become a shared chromosome of the entire human race. It is therefore currently generally accepted that the entire human population evolved from a very small group less than 160,000 years ago, possibly in Africa. Subsequently, through selection pressure, an identifiable gap, not only in form but especially in function, has arisen in modern humankind. The biblical narrative of Adam and Eve meets all the requirements for the threshold event that has been scientifically distinguished. In the Bible, it is said that God created man from the dust of the earth - in His image He created him. That humans have a common ancestor with monkeys seems to be true, just like with every organism on earth. We share much more than just ancestors with the rest of creation as was already stated, also building material, cells, organs, and a large part of our genome. That humans are unique in nature is, however, undeniable, and contemporary science cannot account for many empirical observations in this regard. It is therefore hypothesised that proto-man was created by guided development from the substance of the earth (carbon, hydrogen, nitrogen, oxygen, phosphate and sulphur). Later, a threshold event occurred (as described in Genesis) during which God transformed the proto-man "in His image". God guided human's minds far beyond survival requirements to develop a language-processed, secondary memory, enabling humankind to receive and internalise love, wisdom, and morality. These are unique attributes of God that make humankind the image of God. Of the change of proto-man's mind and his behaviour to modern humankind during the threshold event, there is ample scientific evidence. This could confirm the paradise narrative as a verifiable reality, as does Eve's creation from Adam. Modern scientific observations regarding the origin of the universe, the development of life and the origin of humans can be best understood when they are interpreted within a religious paradigm. Arno Penzias, physicist, and Nobel laureate is one of many scientists who conclude that the scientific evidence is consistent with the expectations flowing from theism. He writes: "The best data we have is exactly what I would have predicted, had I nothing to go on but the five books of Moses, the Psalms, the Bible as a whole."

Volgens materialiste kan die ontstaan van die mens verklaar word deur evolusie, waarvoor daar oortuigende bewyse is, soos bv. genetiese ooreenkomste, bewyse uit die fossielrekord sowel as anatomiese vergelykings. Daarteenoor stel die metafisiese siening dat mense deur 'n goddelike entiteit of 'n bonatuurlike krag geskep is. Hierdie siening word dikwels geassosieer met godsdienstige oortuigings en is (volgens die teenstanders daarvan) nie op empiriese bewyse gebaseer nie. As 'n spesie, toon menslike DNS (deoksiribonukleïensuur) 'n baie laer diversiteit as wat die genetiese teorie voorspel, hoofsaaklik as gevolg van die resentheid van die mens sowel as die voorkoms van 'n "bevolkingsbottelnek". Dit dui daarop dat die hele menslike bevolking minder as 160 000 jaar gelede uit 'n baie klein groepie ontwikkel het, waarskynlik in Afrika. Mense deel veel meer as net voorouers met die res van die skepping: ook boumateriaal, selle, organe en 'n groot deel van ons genoom. Dat mense uniek van aard is, is egter onbetwisbaar en kontemporêre wetenskap lewer vele empiriese waarnemings in hierdie verband, wat nie bevredigend verklaar kan word deur 'n onbegeleide, toevallige ontwikkelingsgeskiedenis nie. Die Bybelse narratief van Adam en Eva voldoen aan al die vereistes vir die drempelgebeurtenis in die menslike geskiedenis, wat wetenskaplik bewysbaar is. Die hipotese word daarom voorgehou dat die proto-mens geskep is deur begeleide ontwikkeling uit die stof van die aarde (koolstof, waterstof, stikstof, suurstof, fosfaat en swael). Later het 'n drempelgebeurtenis plaasgevind waartydens God die proto-mens "na Sy beeld" getransformeer het.

Abstract Introduction: Gene expression studies require extraction protocols that allow obtaining high quality RNA, especially when working with tissues rich in polysaccharides, lipids and polyphenols such as pecan nut (Carya illinoinensis [Wangenh.] K. Koch) embryo tissue. Objective: To evaluate the efficiency of eight methods of total RNA extraction from pecan nut embryo tissue. Materials and methods: Eight total RNA extraction protocols based on TRI Reagent®, CTAB (hexadecyltrimethylammonium bromide) buffer and a commercial kit were evaluated. Total RNA yield and quality were determined by spectrophotometry (UV/visible). RNA viability and integrity were analyzed by RT-PCR using actin as a reference gene. Results and discussion: Extraction protocols based on TRI Reagent® provided high concentrations of total RNA, but with a high degree of contamination. The commercial kit was used to extract total RNA, but without the expected optimal purity. Finally, protocols based on CTAB buffer achieved total RNA yields of optimal quality. Conclusions: The quality of total RNA varies according to the efficiency of the method used. The CTAB 4 protocol represents an efficient alternative for the isolation of RNA from embryonic tissues of C. illinoinensis.

Resumen Introducción: Los estudios de expresión génica requieren protocolos de extracción que permitan la obtención de ARN de alta calidad, especialmente cuando se trabaja con tejidos ricos en polisacáridos, lípidos y polifenoles como el tejido embrionario de nuez pecanera (Carya illinoinensis [Wangenh.] K. Koch). Objetivo: Evaluar la eficiencia de ocho métodos de extracción de ARN total a partir de tejido embrionario de nuez pecanera. Materiales y métodos: Se evaluaron ocho protocolos de extracción de ARN total basados en el reactivo TRI Reagent®, buffer CTAB (bromuro de hexadeciltrimetilamonio) y un kit comercial. El rendimiento y calidad de ARN total se determinaron por espectrofotometría (UV/visible). La viabilidad e integridad del ARN se analizó mediante RT-PCR utilizando actina como gen de referencia. Resultados y discusión: Los protocolos de extracción basados en el reactivo TRI Reagent® permitieron la obtención de concentraciones altas de ARN total, pero con grado elevado de contaminación. Mediante el uso del kit comercial fue posible la extracción de ARN total, pero sin la pureza óptima esperada. Finalmente, los protocolos basados en el buffer CTAB consiguieron rendimientos de ARN total de calidad óptima. Conclusiones: La calidad del ARN total varía de acuerdo con la eficiencia del método utilizado. El protocolo CTAB 4 representa una alternativa eficaz para el aislamiento de ARN de tejidos embrionarios de C. illinoinensis.

ABSTRACT Introduction: Knockdown of fat mass and obesity-associated gene (FTO) can induce N6-methyladenosine (m 6A) ribonucleic acid (RNA) methylation. The objective of this study was to explore the effect of m 6A RNA methylation on atherosclerotic vulnerable plaque by FTO knockdown. Methods: A total of 50 New Zealand white rabbits were randomly divided into pure high-fat group, sham operation group, vulnerable plaque group, empty load group, and FTO knockdown group (10 rabbits/group). Results: Flow cytometry showed that helper T (Th) cells in the FTO knockdown group accounted for a significantly higher proportion of lymphocytes than in the vulnerable plaque group and empty load group (P<0.05). Th cells were screened by cell flow. The level of m 6A RNA methylation in the FTO knockdown group was significantly higher than in the vulnerable plaque group and empty load group (P<0.05). The levels of total cholesterol, triglyceride, and low-density lipoprotein C were higher at the 12th week than at the 1st week, but the high-density lipoprotein C level was lower at the 12th week than at the 1st week. At the 12th week, the interleukin-7 level was significantly lower in the adeno-associated virus-9 (AVV9)-FTO short hairpin RNA group than in the control and AVV9-green fluorescent protein groups (P<0.001). Conclusion: After successfully establishing a vascular parkinsonism rabbit model, m 6A RNA methylation can decrease Th cells and vulnerable atherosclerotic plaques. Introduction obesityassociated obesity associated (FTO N6methyladenosine Nmethyladenosine N6 methyladenosine N (RNA Methods 5 highfat high 10 (1 rabbits/group. rabbitsgroup rabbits/group . rabbits/group) Results (Th P<0.05. P005 P P<0.05 0 05 (P<0.05) flow cholesterol triglyceride lowdensity low density th st highdensity interleukin7 interleukin 7 interleukin- adenoassociated adeno virus9 virus 9 virus- AVV9FTO AVVFTO AVV9 AVV AVV9green AVVgreen green P<0.001. P0001 P<0.001 001 (P<0.001) Conclusion model plaques 1 ( P00 P<0.0 (P<0.05 P000 P<0.00 00 (P<0.001 P0 P<0. (P<0.0 (P<0.00 P<0 (P<0. P< (P<0 (P< (P

Resumo Os conflitos militares têm sido obstáculos significativos na detecção e tratamento de doenças infecciosas devido à diminuição da infraestrutura de saúde pública, resultando na endemicidade da malária. Uma variedade de incidentes violentos e destrutivos foi vivida pelas FATA (áreas tribais administradas pelo governo federal). Foi uma luta buscar uma análise epidemiológica devido ao conflito contínuo e à talibanização. Isolados clínicos foram coletados de agências Bajaur, Mohmand, Khyber e Orakzai, de maio de 2017 a maio de 2018. Para a coloração de Giemsa, amostras de sangue completo com EDTA foram coletadas de participantes sintomáticos. Isolados de microscopia positivos para malária foram colocados em papéis de filtro para futura detecção molecular plasmódica por reação em cadeia da polimerase aninhada (nPCR) de primers específicos de genes de subunidade ribossômica de ácido ribonucleico (ssrRNA). Desde a reconfirmação do nPCR, um estudo malariométrico de 762 pacientes encontrou 679 casos positivos de malária. Plasmodium vivax foi 523 (77%), Plasmodium falciparum 121 (18%), 35 (5%) eram com infecção de espécies mistas (P. vivax mais P. falciparum) e 83 foram declarados negativos por PCR. Entre as cinco agências da FATA, a agência Khyber tem a maior incidência de malária (19%), seguida por P. vivax (19%) e P. falciparum (4,1%). Em contraste, Kurram tem cerca de 14%, incluindo 10,8% casos de P. vivax e 2,7% P. falciparum, a epidemiologia de malária mais baixa. Surpreendentemente, não há diferenças significativas na distribuição da infecção de espécies mistas entre todas as cinco agências. P. falciparum e P. vivax foram duas espécies prevalentes de malária FATA na área devastada pela guerra no Paquistão. Para superar essa incidência crescente de malária, este estudo recomenda que o início de campanhas de conscientização sobre a malária na escola deve ser apoiado por agências de saúde pública e educação relacionada com a malária localmente, visando crianças e pais.

Abstract Military conflicts have been significant obstacles in detecting and treating infectious disease diseases due to the diminished public health infrastructure, resulting in malaria endemicity. A variety of violent and destructive incidents were experienced by FATA (Federally Administered Tribal Areas). It was a struggle to pursue an epidemiological analysis due to continuing conflict and Talibanization. Clinical isolates were collected from Bajaur, Mohmand, Khyber, Orakzai agencies from May 2017 to May 2018. For Giemsa staining, full blood EDTA blood samples have been collected from symptomatic participants. Malaria-positive microscopy isolates were spotted on filter papers for future Plasmodial molecular detection by nested polymerase chain reaction (nPCR) of small subunit ribosomal ribonucleic acid (ssrRNA) genes specific primers. Since reconfirming the nPCR, a malariometric study of 762 patients found 679 positive malaria cases. Plasmodium vivax was 523 (77%), Plasmodium falciparum 121 (18%), 35 (5%) were with mixed-species infection (P. vivax plus P. falciparum), and 83 were declared negative by PCR. Among the five agencies of FATA, Khyber agency has the highest malaria incidence (19%) with followed by P. vivax (19%) and P. falciparum (4.1%). In contrast, Kurram has about (14%), including (10.8%) P. vivax and (2.7%) P. falciparum cases, the lowest malaria epidemiology. Surprisingly, no significant differences in the distribution of mixed-species infection among all five agencies. P. falciparum and P. vivax were two prevalent FATA malaria species in Pakistan’s war-torn area. To overcome this rising incidence of malaria, this study recommends that initiating malaria awareness campaigns in school should be supported by public health agencies and malaria-related education locally, targeting children and parents alike.

Resumo A resistência do Plasmodium falciparum à cloroquina (CQ) é uma causa significativa de mortalidade e morbidade em todo o mundo. Há uma escassez de dados documentados sobre a prevalência de haplótipos mutantes CQ-resistentes dos genes Pfcrt e Pfmdr1 da guerra endêmica da malária em áreas tribais administradas pelo governo federal do Paquistão. O objetivo deste estudo foi investigar a prevalência de resistência a CQ de P. falciparum nesta área. Isolados clínicos foram coletados entre maio de 2017 e maio de 2018 nas agências do Waziristão do Norte e do Waziristão do Sul da Área de Ensaio Administrada Federalmente. Posteriormente, esfregaços de sangue corados com Giemsa foram examinados para detectar Plasmodium falciparum. A extração do DNA da malária foi feita a partir de amostras de P. falciparum positivas para microscopia, e as infecções por P. falciparum foram confirmadas por nested PCR (visando genes de ácido ribonucleico ribossômico de subunidade pequena de Plasmodium (ssrRNA)). Todas as amostras de P. falciparum confirmadas por PCR foram sequenciadas por pirosequenciamento para descobrir a mutação no gene Pfcrt no códon K76T e em pfmdr1 nos códons N86Y, Y184F, N1042D e D1246Y. De 121 microscopias de casos positivos de P. falciparum, 109 amostras foram positivas para P. falciparum por nested PCR. A mutação Pfcrt K76T foi encontrada em 96% dos isolados, a mutação Pfmdr1 N86Y foi observada em 20% e 11% abrigou a mutação Y184F. Todas as amostras eram do tipo selvagem para o códon N1042D e D1246Y de Pfmdr1. No FATA, Paquistão, a frequência do alelo resistente 76T permaneceu alta apesar da remoção de CQ. No entanto, as descobertas atuais do estudo sugerem a fixação completa do genótipo resistente a CQ de P. falciparum na área de estudo.

Abstract Plasmodium falciparum resistance to Chloroquine (CQ) is a significant cause of mortality and morbidity worldwide. There is a paucity of documented data on the prevalence of CQ-resistant mutant haplotypes of Pfcrt and Pfmdr1 genes from malaria-endemic war effected Federally Administered Tribal Areas of Pakistan. The objective of this study was to investigate the prevalence of P. falciparum CQ-resistance in this area. Clinical isolates were collected between May 2017 and May 2018 from North Waziristan and South Waziristan agencies of Federally Administrated Trial Area. Subsequently, Giemsa-stained blood smears were examined to detect Plasmodium falciparum. Extraction of malarial DNA was done from microscopy positive P. falciparum samples, and P. falciparum infections were confirmed by nested PCR (targeting Plasmodium small subunit ribosomal ribonucleic acid (ssrRNA) genes). All PCR confirmed P. falciparum samples were sequenced by pyrosequencing to find out mutation in Pfcrt gene at codon K76T and in pfmdr1 at codons N86Y, Y184F, N1042D, and D1246Y. Out of 121 microscopies positive P. falciparum cases, 109 samples were positive for P. falciparum by nested PCR. Pfcrt K76T mutation was found in 96% of isolates, Pfmdr1 N86Y mutation was observed in 20%, and 11% harboured Y184F mutation. All samples were wild type for Pfmdr1 codon N1042D and D1246Y. In the FATA, Pakistan, the frequency of resistant allele 76T remained high despite the removal of CQ. However, current findings of the study suggest complete fixation of P. falciparum CQ-resistant genotype in the study area.

Abstract Bacteria responsible for causing infections are common in hospital environments, water, soil, and food products. The infection risk is intensified by the absence of public sanitation, poor quality of life, and food scarcity. These external factors promote the dissemination of pathogens by direct contamination or biofilm formation. In this work, we identified bacterial isolates obtained from intensive care units in the southern region of Tocantins, Brazil. We compared matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) techniques and 16S ribosomal ribonucleic acid (rRNA) molecular analysis; we also performed phenotypic characterization. Fifty-six isolates characterized using morphotinctorial tests were classified as gram-positive (80.4%; n = 45) and gram-negative (19.6%; n = 11) and were resistant to several antibiotic classes; notably, we identified the blaOXA-23 resistance gene in the ILH10 isolate. Microbial identification using MALDI-TOF MS resulted in the identification of Sphingomonas paucimobilis and Bacillus circulans. 16S rRNA sequencing revealed four isolates belonging to the genera Bacillus and Acinetobacter. The similarity was superior to 99% for Acinetobacter schindleri in the Basic Local Alignment Search Tool (BLAST), grouped in the clade superior to 90%. Several strains isolated from intensive care units (ICU) were resistant to various antibiotic classes. These techniques allowed for the identification of several microorganisms of importance in public health, enabling improvements in human infection control and proving the quality of inputs, food, and water. environments water soil products sanitation life scarcity formation work Tocantins Brazil matrixassisted matrix assisted timeofflight time flight MALDITOF MALDI TOF S (rRNA analysis characterization Fiftysix Fifty six grampositive gram positive 80.4% 804 80 4 (80.4% 45 gramnegative negative 19.6% 196 19 6 (19.6% 11 classes notably blaOXA23 blaOXA 23 blaOXA-2 ILH ILH1 isolate circulans 99 BLAST, BLAST , (BLAST) 90 90% ICU (ICU health inputs 80.4 8 (80.4 19.6 1 (19.6 blaOXA2 2 blaOXA- 9 (BLAST 80. (80. 19. (19. (80 (19 (8 (1 (

Resumo As bactérias responsáveis por causar infecções são comuns em ambientes hospitalares, água, solo e produtos alimentícios. O risco de infecção é intensificado pela ausência de saneamento público, má qualidade de vida e escassez de alimentos. Esses fatores externos promovem a disseminação de patógenos por contaminação direta ou formação de biofilme. Neste trabalho, identificamos isolados bacterianos obtidos de unidades de terapia intensiva na região sul do Tocantins, Brasil. Comparamos técnicas de espectrometria de massa de tempo de voo com ionização por dessorção a laser assistida por matriz (MALDI-TOF MS) e análise molecular de ácido ribonucleico ribossômico 16S (rRNA); também realizamos caracterização fenotípica. Cinquenta e seis isolados caracterizados por testes morfotintoriais foram classificados como gram-positivos (80,3%; n = 45) e gram-negativos (19,6%; n = 11) e foram resistentes a várias classes de antibióticos; notavelmente, identificamos o gene de resistência blaOXA-23 no isolado de ILH10. A identificação microbiana usando MALDI-TOF MS resultou na identificação de Sphingomonas paucimobilis e Bacillus circulans. O sequenciamento do 16S rRNA revelou quatro isolados pertencentes aos gêneros Bacillus e Acinetobacter. A similaridade foi superior a 99% para Acinetobacter schindleri no BLAST, agrupado no clado superior a 90%. Várias cepas isoladas de ICU foram resistentes a várias classes de antibióticos. Essas técnicas permitiram a identificação de diversos microrganismos de importância em saúde pública, possibilitando melhorias no controle de infecções humanas e comprovando a qualidade dos insumos, alimentos e água. hospitalares água alimentícios público biofilme trabalho Tocantins Brasil MALDITOF MALDI TOF S (rRNA) fenotípica grampositivos gram positivos 80,3% 803 80 3 (80,3% 45 gramnegativos negativos 19,6% 196 19 6 (19,6% 11 antibióticos notavelmente blaOXA23 blaOXA 23 blaOXA-2 ILH10 ILH circulans 99 BLAST 90 90% pública insumos (rRNA 80,3 8 (80,3 4 19,6 1 (19,6 blaOXA2 2 blaOXA- ILH1 9 80, (80, 19, (19, (80 (19 (8 (1 (

ABSTRACT The growth differentiation factor 9 (GDF9) gene plays a vital role in the growth and maturation of ovarian follicles in laying hens. However, its messenger ribonucleic acid (mRNA) expression levels in preovulatory ovarian follicles of indigenous chickens remain poorly understood. The study aimed to identify the association between egg weight and egg quality traits, ovarian follicles morphology, and mRNA expression levels of the GDF9 gene in pre-ovulatory ovarian follicles of the South African Potchefstroom Koekoek chicken breed. The correlation results showed that egg weight (EW) had a positively high significant correlation (p<0.01) with egg width (EWD), yolk weight (YW), shell surface area (SSA), albumen weight (AW), albumen ratio (AR) and egg volume (EV), and a positive significant correlation (p<0.05) with egg length (EL). The Student’s T-test results revealed that the numbers of large yellow follicles were significantly lower (p<0.05) than those of small yellow follicles. ANOVA findings showed that there was a significant difference (p<0.05) in the average weight of the large yellow follicles. The quantitative Reverse Transcription Polymerase Chain Reaction (RT-qPCR) findings indicated that there were significant differences (p<0.05) in the mRNA expression levels of the GDF9 gene in preovulatory ovarian follicles of the Potchefstroom Koekoek chicken breed. The mRNA expression was more abundant in F1 and F4 than in other ovarian follicles. GDF (GDF9 hens However (mRNA understood traits morphology pre ovulatory breed EW (EW p<0.01 p001 p 0 01 (p<0.01 EWD, EWD , (EWD) YW, YW (YW) SSA, SSA (SSA) AW, AW (AW) AR (AR EV, EV (EV) p<0.05 p005 05 (p<0.05 EL. EL . (EL) Students Student s Ttest T test RTqPCR RT qPCR (RT-qPCR F (GDF p<0.0 p00 (p<0.0 (EWD (YW (SSA (AW (EV (EL p<0. p0 (p<0. p<0 (p<0 p< (p< (p

ABSTRACT Introduction: Thoracic aortic aneurysm is a potentially fatal disease with a strong genetic contribution. The dysfunction of vascular smooth muscle cells (VSMCs) contributes to the formation of this aneurysm. Although previous studies suggested that long non-coding ribonucleic acid (RNA) hypoxia inducible factor 1 α-antisense RNA 1 (HIF1A-AS1) exerted a vital role in the progression and pathogenesis of thoracic aortic aneurysm, we managed to find a new regulatory mechanism of HIF1A-AS1 in VSMCs via transcriptomics. Methods: Cell viability was detected by the cell counting kit-8 assay. Cell apoptosis was assessed by Annexin V-fluorescein isothiocyanate/propidium iodide double staining. Transwell migration assay and wound healing assay were performed to check the migration ability of HIF1A-AS1 on VSMCs. The NextSeq XTen system (Illumina) was used to collect RNA sequencing data. Lastly, reverse transcription-quantitative polymerase chain reaction confirmed the veracity and reliability of RNA-sequencing results. Results: We observed that overexpressing HIF1A-AS1 successfully promoted apoptosis, significantly altered cell cycle distribution, and greatly attenuated migration in VSMCs, further highlighting the robust promoting effects of HIF1A-AS1 to thoracic aortic aneurysm. Moreover, transcriptomics was implemented to uncover its underlying mechanism. A total of 175 differently expressed genes were identified, with some of them enriched in apoptosis, migration, and cell cycle-related pathways. Intriguingly, some differently expressed genes were noted in vascular development or coagulation function pathways. Conclusion: We suggest that HIF1A-AS1 mediated the progression of thoracic aortic aneurysm by not only regulating the function of VSMCs, but also altering vascular development or coagulation function. Introduction contribution (VSMCs noncoding non coding (RNA αantisense α antisense HIF1AAS1 HIFAAS HIF1A AS1 HIF AS (HIF1A-AS1 HIF1A-AS Methods kit8 kit 8 kit- Vfluorescein V fluorescein isothiocyanatepropidium isothiocyanate propidium staining Illumina (Illumina data Lastly transcriptionquantitative transcription quantitative RNAsequencing results Results distribution Moreover 17 identified cyclerelated related pathways Intriguingly Conclusion AAS HIF1AAS HIFA (HIF1A-AS

Cowdria polymorphic gene 1 (cpg1, Erum2510, ERUM_RS01380) has been shown to induce 30% and 100% protection in sheep immunised by deoxyribonucleic acid (DNA) prime combined with DNA boost and DNA prime combined with protein boost, respectively, against heartwater infection via needle challenge. To localise its antigenic regions for inclusion in a multi-epitope DNA vaccine against heartwater, Erum2510 was cleaved into five overlapping subfragments. These subfragments were expressed individually in an Escherichia coli host expression system and evaluated for their ability to induce proliferative responses, Th1 and Th2 cytokines (interferon gamma [IFN-γ] and interleukin 4 [IL-4]) via enzyme-linked immunospot (ELISpot), quantitative real time polymerase chain reaction (qRT-PCR) and flow cytometry. Recombinant (r)proteins 3 and 4 were shown to induce immunodominant Th1 and Th2 immune responses characterised by the secretion of effector cytokines IFN-γ and IL-4 in addition to differential messenger ribonucleic acid (mRNA) expression of tumour necrosis factor (TNF), IL-2, IL-1, IL-18, IL-10, transforming growth factor (TGF), granulocyte-macrophage colony-stimulating factor (GM-CSF) and inducible nitric oxide synthase (iNOS). Thirty-seven overlapping synthetic peptides (16 mer) spanning the lengths of these immunodominant rproteins were synthesised and assayed. A peptide pool comprising p9 and p10 derived from rprotein 3 induced a Th1-biased immune response. A peptide pool comprising p28 and p29 derived from rprotein 4 induced a mixed Th1 and Th2 immune response characterised by secretion of IFN-γ and differential mRNA expression of IL-1, IL-2, IL-10, IL-12, iNOS, TGF, TNF and GM-CSF. Only one of the peptides (p29) induced secretion of IL-4. Phenotypic analysis showed significant activation of cluster of differentiation 8+ (CD8+), cluster of differentiation 4+ (CD4+) and B+ lymphocyte populations. Findings suggest that Erum2510 rproteins and synthetic peptides can induce both cellular and humoral immune responses, thereby implicating their importance in protection against heartwater. CONTRIBUTION: This study will facilitate the design of an effective multi-epitope DNA vaccine against heartwater that will contribute to control this economically important disease in sub-Saharan Africa and beyond.

ABSTRACT Background: COVID-19 pandemic situation made the pharmaceutical companies develop the vaccine with different formulations in a short period. Objectives: The main objective of the review is to focus on different types of vaccine formulations available globally and the importance of technology transfer in vaccine development associated with potential risks. Results: Research on vaccine development led to various types of vaccines, such as Inactivated vaccines, Live Attenuated vaccines, Ribonucleic acid (RNA) and Deoxyribonucleic acid (DNA) vaccines, viral vector vaccines, and Protein Subunit Vaccines for COVID-19. But the process of vaccine development and technology transfer is lined with various risks and challenges. Through risk assessment, we found some major potential risks involved in product development; this leads to a smoother and more efficient method to develop safe vaccines available for public health. Conclusions: This review will explain the significance of technology collaboration for the faster development of various formulations of vaccines globally.

RESUMEN Antecedentes: La situación de pandemia de COVID-19 hizo que las empresas farmacéuticas desarrollaran la vacuna con diferentes formulaciones en un corto período. Objetivos: El objetivo principal de la revisión es centrarse en los diferentes tipos de formulaciones de vacunas disponibles a nivel mundial y la importancia de la transferencia de tecnología en el desarrollo de vacunas asociado con los riesgos potenciales. Resultados: La investigación sobre el desarrollo de vacunas condujo al desarrollo de varios tipos de vacunas, como vacunas inactivadas, vacunas vivas atenuadas, vacunas de ácido ribonucleico (ARN) y ácido desoxirribonucleico (ADN), vacunas de vectores virales y vacunas de subunidades de proteínas para COVID-19. Pero el proceso de desarrollo de vacunas y transferencia de tecnología está lleno de varios riesgos y desafíos. A través de la evaluación de riesgos, encontramos algunos riesgos potenciales importantes involucrados en el desarrollo de productos, lo que conduce a un método más fluido y eficiente para desarrollar vacunas seguras disponibles para la salud pública. Conclusiones: Esta revisión dará una idea de la importancia de la colaboración tecnológica para el desarrollo más rápido de varias formulaciones de vacunas a nivel mundial.