Resumo O câncer é uma doença maligna fatal e sua crescente prevalência mundial exige a descoberta de biomarcadores moleculares mais sensíveis e confiáveis. Investigar o nível de expressão de GINS1 e seu valor prognóstico em cânceres humanos distintos, usando uma série de abordagens in silico em várias camadas, pode ajudar a estabelecê-lo como um potencial biomarcador de diagnóstico e prognóstico compartilhado de diferentes subtipos de câncer. O mRNA de GINS1, a expressão da proteína e a metilação do promotor foram analisados usando UALCAN e Human Protein Atlas (HPA), enquanto a expressão de mRNA foi posteriormente validada via GENT2. Os valores prognósticos potenciais de GINS1 foram avaliados por meio do plotter KM. Em seguida, o cBioPortal foi utilizado para examinar as mutações genéticas relacionadas ao GINS1 e as variações do número de cópias (CNVs), enquanto a análise de enriquecimento da via foi realizada usando DAVID. Além disso, uma análise correlacional entre a expressão de GINS1 e células imunes T CD8 + e a construção de uma rede de interação gene-droga foi realizada usando TIMER, CDT e Cytoscape. O GINS1 foi encontrado regulado negativamente em um único subtipo de câncer humano, enquanto comumente regulado positivamente em 23 outros subtipos diferentes. A regulação positiva de GINS1 foi significativamente correlacionada com a sobrevida global pobre (OS) de Carcinoma Hepatocelular de Fígado (LIHC), Adenocarcinoma de Pulmão (LUAD) e Carcinoma de Células Claras Renais de Rim (KIRC). O GINS1 também foi encontrado regulado positivamente em pacientes LIHC, LUAD e KIRC de diferentes características clínico-patológicas. A análise de enriquecimento de vias revelou o envolvimento de GINS1 em duas vias diversas, enquanto poucas correlações interessantes também foram documentadas entre a expressão de GINS1 e seu nível de metilação do promotor, nível de células imunes T CD8 + e CNVs. Além disso, também previmos poucos medicamentos que poderiam ser usados no tratamento de LIHC, LUAD e KIRC, regulando a expressão de GINS1. O perfil de expressão de GINS1 no estudo atual sugeriu que é um novo biomarcador de diagnóstico e prognóstico compartilhado de LIHC, LUAD e KIRC.

Abstract Cancer is a fatal malignancy and its increasing worldwide prevalence demands the discovery of more sensitive and reliable molecular biomarkers. To investigate the GINS1 expression level and its prognostic value in distinct human cancers using a series of multi-layered in silico approach may help to establish it as a potential shared diagnostic and prognostic biomarker of different cancer subtypes. The GINS1 mRNA, protein expression, and promoter methylation were analyzed using UALCAN and Human Protein Atlas (HPA), while mRNA expression was further validated via GENT2. The potential prognostic values of GINS1 were evaluated through KM plotter. Then, cBioPortal was utilized to examine the GINS1-related genetic mutations and copy number variations (CNVs), while pathway enrichment analysis was performed using DAVID. Moreover, a correlational analysis between GINS1 expression and CD8+ T immune cells and a the construction of gene-drug interaction network was performed using TIMER, CDT, and Cytoscape. The GINS1 was found down-regulated in a single subtypes of human cancer while commonly up-regulated in 23 different other subtypes. The up-regulation of GINS1 was significantly correlated with the poor overall survival (OS) of Liver Hepatocellular Carcinoma (LIHC), Lung Adenocarcinoma (LUAD), and Kidney renal clear cell carcinoma (KIRC). The GINS1 was also found up-regulated in LIHC, LUAD, and KIRC patients of different clinicopathological features. Pathways enrichment analysis revealed the involvement of GINS1 in two diverse pathways, while few interesting correlations were also documented between GINS1 expression and its promoter methylation level, CD8+ T immune cells level, and CNVs. Moreover, we also predicted few drugs that could be used in the treatment of LIHC, LUAD, and KIRC by regulating the GINS1 expression. The expression profiling of GINS1 in the current study has suggested it a novel shared diagnostic and prognostic biomarker of LIHC, LUAD, and KIRC.

Resumo Acacia modesta (AM) e Opuntia monocantha (OM) estão distribuídas no Paquistão, Afeganistão e Índia. Ambas as plantas têm propriedades farmacológicas diferentes. Este estudo foi desenhado para avaliar o potencial anticancerígeno de Acacia modesta (AM) e Opuntia monocantha (OM). A linha celular de câncer de fígado HepG2 foi usada para avaliação da atividade anticâncer. Para a avaliação dos efeitos antiproliferativos, a viabilidade celular e a morte celular em todos os grupos de células foram avaliadas através dos ensaios de MTT, cristal violeta e azul de tripano. Para a avaliação da apoptose foi realizado ELISA de p53. Além disso, o ensaio de LDH para descobrir a capacidade das células malignas de metabolizar o piruvato em lactato e a atividade das enzimas antioxidantes (GSH, CAT e SOD) no final da HPLC foi realizado para encontrar o composto ativo de AM e OM. Citotoxicidade (MTT), ensaios de viabilidade (azul de tripano, viabilidade de cristal, análise MUSE) mostraram mais células mortas e menos células vivas nos grupos tratados com plantas com aumento da concentração. O ensaio de arranhão para o efeito anti-migratório dessas plantas mostrou que os grupos tratados não têm capacidade de cicatrizar arranhão/ferida. ELISA de p53 para apoptose celular mostrou maior liberação de p53 nos grupos tratados. Ensaio antioxidante via glutationa (GSH), superóxido dismutase (SOD), catalase (CAT) mostrou menor potencial antioxidante nos grupos de câncer tratados. O ensaio de LDH mostrou mais liberação de lactato desidrogenase nos grupos tratados em comparação com os não tratados. A análise de HPLC mostrou a presença de fitoquímicos como esteróides, alcalóides, fenóis, flavonóides, saponinas, taninos, antraquinona e aminoácidos nos extratos vegetais AM e OM. Com base em todos esses achados, pode-se concluir que os extratos etanólicos de Acacia modesta e Opuntia monocantha apresentam potencial anticancerígeno promissor.

Abstract Acacia modesta (AM) and Opuntia monocantha (OM) are distributed in Pakistan, Afghanistan and India. Both of these plants have different pharmacological properties. This study was designed to evaluate anticancer potential of Acacia modesta (AM) and Opuntia monocantha (OM). Liver cancer cell line HepG2 was used for assessment of anticancer activity. For the evaluation of anti-proliferative effects, cell viability and cell death in all groups of cells were evaluated via MTT, crystal violet and trypan blue assays. For the evaluation of apoptosis ELISA of p53 performed. Furthermore, LDH assay to find out the ability of malignant cells to metabolize pyruvate to lactate and antioxidant enzymes activity (GSH, CAT and SOD) at the end HPLC was performed to find active compound of AM and OM. Cytotoxicity (MTT), Viability assays (trypan blue, crystal viability, MUSE analysis) showed more dead, less live cells in plant treated groups with increase of concentration. Scratch assay for the anti-migratory effect of these plants showed treated groups have not ability to heal scratch/wound. ELISA of p53 for cellular apoptosis showed more release of p53 in treated groups. Antioxidant assay via glutathione (GSH), superoxide dismutase (SOD), catalase (CAT) showed less anti-oxidative potential in treated cancer groups. LDH assay showed more lactate dehydrogenase release in treated groups compared with untreated. HPLC analysis showed the presence of phytochemicals such as steroids, alkaloids, phenols, flavonoids, saponins, tannins, anthraquinone and amino acids in AM and OM plant extracts. Based on all these findings, it can be concluded that ethanolic extracts of Acacia modesta and Opuntia monocantha have promising anti-cancer potential.

Resumo Mutações de ocorrência natural na proteína morfogenética 15 (BMP15) estão associadas à diminuição da taxa de ovulação (TO), tamanho da ninhada (TN) e esterilidade. Estudar a BMP15 na raça Cholistani para elevar o status socioeconômico e o conhecimento da criação de ovinos Cholistani no sul de Punjab, Paquistão. Em nosso estudo, 50 ovelhas Cholistani inférteis sem parentesco sanguíneo foram rastreadas para mutações BMP15. Para tanto, um DNA de alta qualidade foi extraído do sangue dessas ovelhas, seguido de concepção do primer, amplificação da reação em cadeia da polimerase (PCR), sequenciamento de DNA e análises in silico. Do total de 50 amostras, 9, incluindo caso 1 (T3), caso 2 (T8), caso 3 (T17), caso 4 (T22), caso 5 (T25), caso 6 (T33), caso 7 (T40), caso 8 (T44) e caso 9 (T47), foram consideradas positivas para uma variedade de mutações BMP15 novas e já relatadas. Mais análises in silico das mutações observadas mostraram o impacto funcional dessas mutações em diferentes características (peso molecular, PI teórico, meia-vida estimada, índice de instabilidade, localização subcelular e confirmação 3D) das proteínas codificadas, possivelmente alterando a funcionalidade normal. Nossos achados confirmaram o possível papel essencial das mutações BMP15 na infertilidade de ovelhas Cholistani.

Abstract Naturally occurring mutations in morphogenetic protein 15 (BMP15) are associated with decreased ovulation rate (OR), litter size (LS), and sterility. It is of a great interest to elucidate BMP15 gene in Cholistani sheep breed to uplift socio-economic status and the knowledge of Cholistani sheep breeding in Southern Punjab, Pakistan. In our study, a total of 50 infertile Cholistani sheep aged between 2-6 years and having no blood relation were screened for BMP15 mutations. For this purpose, a high-quality DNA was extracted from the blood of sheep followed by primer designing, Polymerase Chain Reaction (PCR) amplification, DNA sequencing, and in silico analyses. Out of total 50 samples, 9 samples including case 1 (T3), case 2 (T8), case 3 (T17), case 4 (T22), case 5 (T25), case 6 (T33), case 7 (T40), case 8 (T44), and case 9 (T47) were found positive for a variety of already reported and novel BMP15 mutations. Further in silico analyses of the observed mutations have shown the functional impact of these mutations on different characteristics (molecular weight, theoretical PI, estimated half-life, instability index, sub-cellular localization, and 3D confirmation) of the encoded proteins, possibly altering the normal functionality. In a nutshell, findings of this study have confirmed the possible essential role of the BMP15 mutations in the infertility of the Cholistani sheep.

Abstract The study attempted to assess preparatory year students’ perception towards pharmacists and the pharmacy profession. This cross-sectional survey was conducted between December 2019 and March 2020. The students were invited to complete an anonymous questionnaire via Google Forms®. In total, 244 students, of which 53.7% were female with the mean age of 19.2 ± 0.65, from 12 universities participated in this study. As per our findings, the majority of the respondents (91.8%) regard pharmacy as a well-respected profession, 82.4% thought pharmacists are important decision-makers, 68.4% disagreed that most pharmacists were unkind, and 60.7% did not agree that pharmacy was a low-status occupation. Meanwhile, 95.5% agreed that pharmacists must have a university degree, 88.6% agreed pharmacists must take responsibility for patients, and 82.8% believed pharmacists had to work too hard. Moreover, 62.3% did not think pharmacy was a low-skill occupation, 54.9% did not agree pharmacists must do unpleasant things, and 45.1% disagreed pharmacists only did what physicians requested of them. Lastly, 48.8% had low confidence in choosing pharmacy as a career. The students’ overall perception toward pharmacists and the pharmacy profession was favorable. However, only one-fourth of the students displayed a tendency to choose pharmacy as a future career.

RESUMO: Foi realizada uma avaliação do potencial antifúngico da parte aérea da gramínea alelopática Cenchrus pennisetiformis (Hochst. & Steud.) Wipff. contra Fusarium oxysporum f. sp. lycopersici Snyder & Hansen, um patógeno fúngico que causa a doença de murcha no tomate (Solanum lycopersicum L.). Diferentes concentrações (1% a 6%) do extrato metanólico da folha, do caule e da inflorescência da gramínea reduziram significativamente a biomassa fúngica em 40-88%, 13-89% e 26-76%, respectivamente. O extrato metanólico (folha + caule) foi fracionado, utilizando-se quatro solventes orgânicos: n-hexano, clorofórmio, acetato de etila e n-butanol. Todas as subfrações de extrato metanólico da parte aérea apresentaram significativo potencial antifúngico em graus variáveis. Concentrações diferentes (1,56-200 mg mL-1) da subfração do acetato de etila exibiram a melhor atividade antifúngica, com supressão de 49-100% na biomassa fúngica. A análise CG-EM da subfração do acetato de etila revelou a presença de 10 compostos. O principal composto (30,99%) foi o fenol, 2,4-bis{1,1-dimetiletil}, seguido de ácido hexadecanoico, éster etílico (21,72%), 2,3-di-hidrobenzofurano (10,65%), 1-propanol-2-2-hidroxipropil (10,60%) e 1-eicoseno (8,32%).

ABSTRACT: Antifungal potential of aerial parts of an allelopathic grass Cenchrus pennisetiformis (Hochst. & Steud.) Wipff. was evaluated against Fusarium oxysporum f. sp. lycopersici Snyder & Hansen, a fungal pathogen causing wilt disease in tomato (Solanum lycopersicum L.). Different concentrations (1% to 6%) of methanolic leaf, stem and inflorescence extract of the grass significantly reduced fungal biomass by 40-88%, 13-89%, and 26-76%, respectively. Methanolic shoot (leaf + stem) extract was fractionated using four organic solvents viz. n-hexane, chloroform, ethyl acetate and n-butanol. All the sub-fractions of methanolic shoot extract showed remarkable antifungal potential to variable extents. Different concentrations (1.56-200 mg mL-1) of ethyl acetate sub-fraction exhibited the best antifungal activity resulting in 49-100% suppression in the fungal biomass. GC-MS analysis of ethyl acetate sub-fraction showed the presence of 10 compounds. Phenol, 2,4-bis{1,1-dimethlethyl}- was the major compound (30.99%) followed by hexadecanoic acid, ethyl-ester (21.72%), benzofuran 2,3-dihydro (10.65%), 1-propanol-2-2-hydroxypropxy (10.60%) and 1-eicosene (8.32%).

RESUMO O presente estudo foi conduzido para avaliar o potencial antifúngico da grama alelopática Cenchrus pennisetiformis Hochst. & Steud. contra o fungo patógeno de plantas Fusarium oxysporum f. sp. lycopersici (causador da doença murcha-de-fusário) sob estresse de cromo. Experimentos laboratoriais foram realizados em tubos de ensaio de vidro com volume de 10 mL, cada um contendo 1,0 mL de extrato de caldo de malte em sete concentrações (50, 100, 150, 200, 250, 300 e 350 ppm), para cada Cr(III) e Cr(VI), e duas concentrações de extrato de folha, caule ou raiz metanólica (5% e 6%) de C. pennisetiformis. Um meio alterado de metal + extrato de grama foi inoculado com o patógeno e incubado por sete dias em uma temperatura de 25 ºC. Concentrações diferentes de Cr(III) e CR(VI) demonstraram resultados igualmente inibitórios em reduções de 1084% e 18-89% na biomassa do fungo, respectivamente, em relação ao controle. Extratos metanólicos de folha, caule e raiz de grama reduziram a biomassa do fungo em 12-25%, 14-23% e 46-50%, respectivamente, quando comparados ao controle. Em aplicações combinadas de extratos metanólicos de partes diferentes de C. pennisetiformis e soluções metais, extratos de raiz combinados tanto com Cr(III) quanto com Cr(VI) mostraram os maiores potenciais inibitórios contra o fungo, seguidos pelos extratos de folha e caule. Na combinação com extratos metanólicos de raiz, folha e caule, concentrações diferentes de Cr(III) e Cr(VI) reduziram significativamente a biomassa do fungo em 59-99%, 14-99% e 9-95%, respectivamente, comparando-se ao controle. Esses estudos não foram realizados anteriormente. Os resultados desta pesquisa sugerem que F. oxysporum f. sp. lycopersici, causador da murcha-do-fusário em tomates, pode ser manejado pela aplicação de extratos (ou, alternativamente, biomassa) de C. pennisetiformis em solos contaminados com cromo.

ABSTRACT The present study was conducted to assess antifungal potential of the allelopathic grass Cenchrus pennisetiformis Hochst. & Steud. against the fungal plant pathogen Fusarium oxysporum f. sp. lycopersici (cause of tomato wilt disease) under chromium stress. Laboratory experiments were performed in 10 mL volume glass test tubes each containing 1.0 mL of malt extract broth with seven concentrations (0, 50, 100, 150, 200, 250, 300 and 350 ppm) of each of Cr(III) and Cr(VI), and two concentrations (5% and 6%) of methanolic leaf, stem or root extract of C. pennisetiformis. A metal + weed extract amended medium was inoculated with the pathogen and incubated for 7 days at 25 C. Different concentrations of Cr(III) and Cr(VI) proved equally inhibitory resulting in 10-84% and 18-89% reduction in fungal biomass, respectively. Methanolic leaf, stem and root extracts of the weed reduced fungal biomass by 12-25%, 14-23% and 46-50%, respectively, over negative control. In combined application of methanolic extracts of different parts of C. pennisetiformis and metal solutions, root extract in combination with either Cr(III) or Cr(VI) showed the highest inhibitory potential against the fungus followed by leaf and stem extracts. In combination with methanolic root, leaf and stem extracts, different concentrations of Cr(III) and Cr(VI) significantly reduced fungal biomass by 54-99%, 14-99% and 9-95%, respectively, over negative control. Such studies have not been carried out previously. Results of the present investigation suggest that F. oxysporum oxysporum f. sp. lycopersici, the cause of Fusarium wilt disease in tomato, can be managed by application of extracts (or alternatively biomass) of C. pennisetiformis in chromium contaminated soils.

Introduction: The treatment of large renal stones in children can be challenging often requiring combination therapy and multiple procedures. The purpose of this video is to describe our technique of robotic nephrolithotomy and pyelolithotomy for complex renal stone disease in children, and to demonstrate the utility of the robotic ultrasound probe to aid with stone localization. Materials and Methods: Robotic nephrolithotomy/pyelolithotomy was carried out in four consecutive patients. A robotic ultrasound probe (Hitachi-Aloka, Tokyo, Japan) under console surgeon control was used in all cases. Results: Two patients underwent robotic pyelolithotomy, one patient underwent robotic nephrolithotomy, whilst the fourth patient underwent robotic pyelolithotomy and nephrolithotomy along with Y-V pyeloplasty for concurrent ureteropelvic junction obstruction. Mean operative time, blood loss and hospital stay was 216 minutes, 37.5 mL and 2 days, respectively. The robotic ultrasound probe aided identification of calculi within the kidney in all cases. For nephrolithotomy it was helpful in planning the incision for nephrotomy. After nephrotomy or pyelotomy, stones were removed using a combination of robotic Maryland forceps, fenestrated grasper or Prograsp. Antegrade nephroscopy introduced through a laparoscopic port was used in all patients for confirmation of residual stone status. Two patients did not require a ureteral stent in the post-operative period. One patient had a minor complication (Clavien Grade 2 - dislodged malecot catheter). All patients were stone free at last follow-up. Conclusions: Robotic nephrolithotomy and pyelolithotomy with utilization of the robotic ultrasound probe offers a one-stop solution for complex renal stones with excellent stone-free rates.