Biodiversity, distributions and isolation of microplastics pollution in finfish species in the Panjkora River at Lower and Upper Dir districts of Khyber Pakhtunkhwa province of Pakistan / Biodiversidade, distribuição e isolamento de poluições por microplásticos em espécies de peixes no rio Panjkora nos distritos de Lower e Upper Dir da província de Khyber Pakhtunkhwa do Paquistão

The basic aim of this study was aimed to determine the ichthyofaunal diversity of River Panjkora in both upper and lower Dir districts in Khyber Pakhtunkhwa province of Pakistan.Fish samples were collected by using fishnets from March to September 2020. A total of 724 specimens were collected and classified into 5 families, 14 genera, and 18 species. The overall results revealed that most fish fauna of river Panjkora contains 8 species of family Cyprinidae (56.49%) followed by 4 species of Nemacheilidae (24.44%), 2 species of Channidae (10.63%), and Sisoridae (7.04%), and 1 species of Mastacembelidae (1.38%), respectively. Among all kinds of fish species, Schizothorax plagiostomus (16.57%) was highly dominated and followed by Carassius auratus (11.87%) and Racoma labiata (9.66%) and were reported as highly abundant, especially during April, May, and June. The least abundant species were Glyptothorax punjabensis, Glyptothorax sufii, and Mastacembelus armatus, that constituting 2.48%, 2.20%, and 1.38% of the total fish samples. The Overall Simpson's diversity (1-D= 0.919) and Simpson's Reciprocal index values (1/D= 12.3876), and Shannon's index (H= 2.68) were indicating that river Panjkora contains a quite rich and diverse group of fish species. The highest microplastics observed in site 7 compared to other study area. Conservation steps should be taken as a top priority to protect and conserve the marine environment and natural heritage from further loss, extinction and stop or minimize losses incurred through irresponsible fishery practices

O objetivo básico deste estudo foi determinar a diversidade ictiofaunística do rio Panjkora nos distritos de Lower e Upper de Dir, na província de Khyber Pakhtunkhwa, no Paquistão. Amostras de peixes foram coletadas com redes de arrasto de março a setembro de 2020. Foram coletados 724 espécimes, classificados em 5 famílias, 14 gêneros e 18 espécies. Os resultados gerais revelaram que a maioria da ictiofauna do rio Panjkora contém 8 espécies da família Cyprinidae (56,49%), seguidas por 4 espécies de Nemacheilidae (24,44%), 2 espécies de Channidae (10,63%) e Sisoridae (7,04%) e 1 espécie de Mastacembelidae (1,38%). Entre todas as espécies de peixes, Schizothorax plagiostomus (16,57%) foi altamente predominante, seguida por Carassius auratus (11,87%) e Racoma labiata (9,66%), e elas foram relatadas como altamente abundantes, especialmente nos meses de abril, maio e junho. As espécies menos abundantes foram Glyptothorax punjabensis, Glyptothorax sufii e Mastacembelus armatus, que constituíram 2,48%, 2,20% e 1,38%, respectivamente, do total de peixes amostrados. O índice de diversidade de Simpson (1-D = 0,919), o índice recíproco de Simpson (1/D = 12,3876) e o índice de Shannon (H = 2,68) indicaram que o rio Panjkora contém um grupo bastante rico e diversificado de espécies de peixes. Os microplásticos mais altos foram observados no local 7 em comparação com outra área de estudo. Medidas de conservação devem ser tomadas como prioridade máxima para proteger e conservar o ambiente marinho e o patrimônio natural de novas perdas e extinção e para parar ou minimizar as perdas ocorridas por práticas de pesca irresponsáveis.

Growth performance and survivability of the Asian seabass Lates calcarifer (Bloch, 1790) reared under hyper-saline, hypo-saline and freshwater environments in a closed aquaculture system / Desempenho de crescimento e sobrevivência do robalo asiático Lates calcarifer (Bloch, 1790) criado em ambientes hipersalinos, hipossalinos e de água doce em um sistema de aquicultura fechado

Salinity is one of the most critical environmental parameters regarding fish physiology, modifying food intake and growth performance in many fish species. The present study has investigated the effects of different salinity levels on growth performance, feeding and survival of Asian seabass Lates calcarifer juveniles. Asian seabass juveniles were reared at 0 (T1), 5 (T2), 22 (T3), 36 (T4), and 42 (T5) ppt salinity. Approximately eight hundred thirty fish individuals with an average weight of 1.24±0.52 g were randomly distributed (166 fish/Tank) in 5 concrete tanks (each tank 30×6×4 ft, volume 19,122 L) for forty days. Juveniles were initially fed 42% crude protein-containing diets at a rate of 6% of their body weight per day. The results showed that salinity level had a significant effect on the weight gain (WG), average daily weight gain (ADWG), specific growth rate (SGR), feed conversion ratio (FCR), survival rate (SR), total biomass and health indices (p<0.05). The highest WG (39.11±1.49 g), ADWG (1.00±0.12 g), SGR (8.74±0.03% d-1) and lowest FCR (0.96±0.20) were observed with T3 treatment, which was significantly higher compared to other treatment groups (p<0.05). Among the health indices, the highest hepatosomatic index and viscerosomatic index were found with T3 treatment, significantly higher than the other groups (p<0.05). No significant differences were found among the treatments in terms of survival rate (p>0.05), but the maximum survival rate (98.89±0.0%) was observed in the T3 and T2 treatments. The maximum level of crude proteins (19.99±1.4%) was found in the whole-body biochemical composition of Asian seabass juveniles in the T3 treatment group. The second-order polynomial regression showed that 20 ppt salinity is optimum for the best growth of Asian seabass. Thus, the present study recommends 20 to 36 ppt salinity for the commercial farming of Asian seabass under a closed aquaculture system.

A salinidade é um dos parâmetros ambientais mais críticos em relação à fisiologia dos peixes, modificando a ingestão de alimentos e o desempenho de crescimento em muitas espécies. O presente estudo investigou os efeitos de diferentes níveis de salinidade no desempenho de crescimento, alimentação e sobrevivência de jovens espécies de robalos asiáticos Lates calcarifer, as quais foram criadas sob salinidade 0 (T1), 5 (T2), 22 (T3), 36 (T4) e 42 (T5) ppt. Aproximadamente 830 indivíduos de peixes com peso médio de 1,24 ± 0,52 g foram distribuídos aleatoriamente (166 peixes / tanque) em cinco tanques de concreto (cada tanque tinha 30 × 6 × 4 pés (9,1 x 1,8 x 1,2 metros), com volume de 19,12 litros) por 40 dias. Os peixes foram inicialmente alimentados com uma dieta contendo 42% de proteína bruta a uma taxa de 6% do seu peso corporal por dia. Os resultados mostraram que o nível de salinidade teve um efeito significativo no ganho de peso (GP), ganho de peso médio diário (GPMD), taxa de crescimento específico (TCE), taxa de conversão alimentar (TCA), taxa de sobrevivência (TS), biomassa total e índices de saúde (p < 0,05). O maior GP (39,11 ± 1,49 g), GPMD (1,00 ± 0,12 g), TCE (8,74 ± 0,03% d-1) e o menor TCA (0,96 ± 0,20) foram observados com o tratamento T3, que foi significativamente superior em comparação com os outros tratamentos (p < 0,05). Entre os índices de saúde, os maiores índices hepatossomáticos e viscerossomáticos foram encontrados no tratamento T3, significativamente superior do que os demais grupos (p < 0,05). Não foram encontradas diferenças significativas entre os tratamentos quanto à TS (p > 0,05), mas a TS máxima (98,89 ± 0,0%) foi observada nos tratamentos T3 e T2. O nível máximo de proteína bruta (19,99 ± 1,4%) foi encontrado na composição bioquímica dos corpos dos jovens robalos asiáticos no grupo T3. A regressão polinomial de segunda ordem mostrou que a salinidade de 20 ppt é ótima para o melhor crescimento do robalo asiático. Assim, o presente estudo recomenda salinidade de 20 a 36 ppt para o cultivo comercial de robalo asiático em sistema de aquicultura fechado.

Study of some morphometric and meristic characteristics of Alepes vari (Cuvier, 1833) collected from the Arabian coast / Estudo de características morfométricas e merísticas de Alepes vari (Cuvier, 1833) coletados na costa da Arábia

The present study examines the correlations between fifteen morphometric and ten meristic characters and total length (TL) of males, females, and combined sexes of Alepes vari (Cuvier, 1833) collected from Karachi fish harbor, West Wharf of Karachi Coast. Statistical analyses of linear regression relationships show mostly strong correlations (r≥0.70; p<0.05) between total length (TL) and most morphometric characters in males, females, and combined sexes, except the height of pectoral-fin (PFH), and pelvic-fin base length (PelFL); whereas, meristic characters were found to be constant and indicate weak or negative type correlations (r≤0.50; p>0.05) with total length (TL). Hence, according to our present results, there is a direct relationship between the total length of fish and all morphometric characters, which were found to be the best indicators of positive allometric pattern growth in fish. Moreover, analysis of the 2-sample t-test revealed (t-test; p>0.05) that no sexual dimorphism was reported in Alepes vari. Thus, our present study could be valuable in systematic classification, sexual dimorphism, and management of this species on the Karachi coast.

O presente estudo examina as correlações entre 15 caracteres morfométricos e 10 caracteres merísticos e comprimento total (CT) de machos, fêmeas e sexos combinados de Alepes vari (Cuvier, 1833), coletados do porto de Karachi, West Wharf, na costa de Karachi. As análises estatísticas das relações de regressão linear mostraram, principalmente, correlações fortes (r ≥ 0,70; p < 0,05) entre o CT e a maioria dos caracteres morfométricos em machos, fêmeas e sexos combinados, exceto a altura da nadadeira peitoral e o comprimento da base da nadadeira pélvica, enquanto os caracteres merísticos foram constantes, indicando correlações fracas ou negativas (r ≤ 0,50; p > 0,05) com o CT. Portanto, de acordo com nossos resultados, existe uma relação direta entre o CT dos peixes e todos os caracteres morfométricos, que foram considerados os melhores indicadores de crescimento do padrão alométrico positivo em peixes. Além disso, a análise do teste t de duas amostras revelou (teste t; p > 0,05) que nenhum dimorfismo sexual foi relatado em A. vari.

Study of some morphometric and meristic characteristics of Alepes vari (Cuvier, 1833) collected from the Arabian coast

Abstract The present study examines the correlations between fifteen morphometric and ten meristic characters and total length (TL) of males, females, and combined sexes of Alepes vari (Cuvier, 1833) collected from Karachi fish harbor, West Wharf of Karachi Coast. Statistical analyses of linear regression relationships show mostly strong correlations (r0.70; p 0.05) between total length (TL) and most morphometric characters in males, females, and combined sexes, except the height of pectoral-fin (PFH), and pelvic-fin base length (PelFL); whereas, meristic characters were found to be constant and indicate weak or negative type correlations (r0.50; p>0.05) with total length (TL). Hence, according to our present results, there is a direct relationship between the total length of fish and all morphometric characters, which were found to be the best indicators of positive allometric pattern growth in fish. Moreover, analysis of the 2-sample t-test revealed (t-test; p>0.05) that no sexual dimorphism was reported in Alepes vari. Thus, our present study could be valuable in systematic classification, sexual dimorphism, and management of this species on the Karachi coast.

Resumo O presente estudo examina as correlações entre 15 caracteres morfométricos e 10 caracteres merísticos e comprimento total (CT) de machos, fêmeas e sexos combinados de Alepes vari (Cuvier, 1833), coletados do porto de Karachi, West Wharf, na costa de Karachi. As análises estatísticas das relações de regressão linear mostraram, principalmente, correlações fortes (r 0,70; p 0,05) entre o CT e a maioria dos caracteres morfométricos em machos, fêmeas e sexos combinados, exceto a altura da nadadeira peitoral e o comprimento da base da nadadeira pélvica, enquanto os caracteres merísticos foram constantes, indicando correlações fracas ou negativas (r 0,50; p > 0,05) com o CT. Portanto, de acordo com nossos resultados, existe uma relação direta entre o CT dos peixes e todos os caracteres morfométricos, que foram considerados os melhores indicadores de crescimento do padrão alométrico positivo em peixes. Além disso, a análise do teste t de duas amostras revelou (teste t; p > 0,05) que nenhum dimorfismo sexual foi relatado em A. vari. Assim, o presente estudo pode ser valioso na classificação sistemática, dimorfismo sexual e manejo dessa espécie na costa de Karachi.

Growth performance and survivability of the Asian seabass Lates calcarifer (Bloch, 1790) reared under hyper-saline, hypo-saline and freshwater environments in a closed aquaculture system

Abstract Salinity is one of the most critical environmental parameters regarding fish physiology, modifying food intake and growth performance in many fish species. The present study has investigated the effects of different salinity levels on growth performance, feeding and survival of Asian seabass Lates calcarifer juveniles. Asian seabass juveniles were reared at 0 (T1), 5 (T2), 22 (T3), 36 (T4), and 42 (T5) ppt salinity. Approximately eight hundred thirty fish individuals with an average weight of 1.24±0.52 g were randomly distributed (166 fish/Tank) in 5 concrete tanks (each tank 30×6×4 ft, volume 19,122 L) for forty days. Juveniles were initially fed 42% crude protein-containing diets at a rate of 6% of their body weight per day. The results showed that salinity level had a significant effect on the weight gain (WG), average daily weight gain (ADWG), specific growth rate (SGR), feed conversion ratio (FCR), survival rate (SR), total biomass and health indices (p 0.05). The highest WG (39.11±1.49 g), ADWG (1.00±0.12 g), SGR (8.74±0.03% d-1) and lowest FCR (0.96±0.20) were observed with T3 treatment, which was significantly higher compared to other treatment groups (p 0.05). Among the health indices, the highest hepatosomatic index and viscerosomatic index were found with T3 treatment, significantly higher than the other groups (p 0.05). No significant differences were found among the treatments in terms of survival rate (p>0.05), but the maximum survival rate (98.89±0.0%) was observed in the T3 and T2 treatments. The maximum level of crude proteins (19.99±1.4%) was found in the whole-body biochemical composition of Asian seabass juveniles in the T3 treatment group. The second-order polynomial regression showed that 20 ppt salinity is optimum for the best growth of Asian seabass. Thus, the present study recommends 20 to 36 ppt salinity for the commercial farming of Asian seabass under a closed aquaculture system.

Resumo A salinidade é um dos parâmetros ambientais mais críticos em relação à fisiologia dos peixes, modificando a ingestão de alimentos e o desempenho de crescimento em muitas espécies. O presente estudo investigou os efeitos de diferentes níveis de salinidade no desempenho de crescimento, alimentação e sobrevivência de jovens espécies de robalos asiáticos Lates calcarifer, as quais foram criadas sob salinidade 0 (T1), 5 (T2), 22 (T3), 36 (T4) e 42 (T5) ppt. Aproximadamente 830 indivíduos de peixes com peso médio de 1,24 ± 0,52 g foram distribuídos aleatoriamente (166 peixes / tanque) em cinco tanques de concreto (cada tanque tinha 30 × 6 × 4 pés (9,1 x 1,8 x 1,2 metros), com volume de 19,12 litros) por 40 dias. Os peixes foram inicialmente alimentados com uma dieta contendo 42% de proteína bruta a uma taxa de 6% do seu peso corporal por dia. Os resultados mostraram que o nível de salinidade teve um efeito significativo no ganho de peso (GP), ganho de peso médio diário (GPMD), taxa de crescimento específico (TCE), taxa de conversão alimentar (TCA), taxa de sobrevivência (TS), biomassa total e índices de saúde (p 0,05). O maior GP (39,11 ± 1,49 g), GPMD (1,00 ± 0,12 g), TCE (8,74 ± 0,03% d-1) e o menor TCA (0,96 ± 0,20) foram observados com o tratamento T3, que foi significativamente superior em comparação com os outros tratamentos (p 0,05). Entre os índices de saúde, os maiores índices hepatossomáticos e viscerossomáticos foram encontrados no tratamento T3, significativamente superior do que os demais grupos (p 0,05). Não foram encontradas diferenças significativas entre os tratamentos quanto à TS (p > 0,05), mas a TS máxima (98,89 ± 0,0%) foi observada nos tratamentos T3 e T2. O nível máximo de proteína bruta (19,99 ± 1,4%) foi encontrado na composição bioquímica dos corpos dos jovens robalos asiáticos no grupo T3. A regressão polinomial de segunda ordem mostrou que a salinidade de 20 ppt é ótima para o melhor crescimento do robalo asiático. Assim, o presente estudo recomenda salinidade de 20 a 36 ppt para o cultivo comercial de robalo asiático em sistema de aquicultura fechado.

Biodiversity, distributions and isolation of microplastics pollution in finfish species in the Panjkora River at Lower and Upper Dir districts of Khyber Pakhtunkhwa province of Pakistan

Abstract The basic aim of this study was aimed to determine the ichthyofaunal diversity of River Panjkora in both upper and lower Dir districts in Khyber Pakhtunkhwa province of Pakistan.Fish samples were collected by using fishnets from March to September 2020. A total of 724 specimens were collected and classified into 5 families, 14 genera, and 18 species. The overall results revealed that most fish fauna of river Panjkora contains 8 species of family Cyprinidae (56.49%) followed by 4 species of Nemacheilidae (24.44%), 2 species of Channidae (10.63%), and Sisoridae (7.04%), and 1 species of Mastacembelidae (1.38%), respectively. Among all kinds of fish species, Schizothorax plagiostomus (16.57%) was highly dominated and followed by Carassius auratus (11.87%) and Racoma labiata (9.66%) and were reported as highly abundant, especially during April, May, and June. The least abundant species were Glyptothorax punjabensis, Glyptothorax sufii, and Mastacembelus armatus, that constituting 2.48%, 2.20%, and 1.38% of the total fish samples. The Overall Simpsons diversity (1-D= 0.919) and Simpsons Reciprocal index values (1/D= 12.3876), and Shannons index (H= 2.68) were indicating that river Panjkora contains a quite rich and diverse group of fish species. The highest microplastics observed in site 7 compared to other study area. Conservation steps should be taken as a top priority to protect and conserve the marine environment and natural heritage from further loss, extinction and stop or minimize losses incurred through irresponsible fishery practices

Resumo O objetivo básico deste estudo foi determinar a diversidade ictiofaunística do rio Panjkora nos distritos de Lower e Upper de Dir, na província de Khyber Pakhtunkhwa, no Paquistão. Amostras de peixes foram coletadas com redes de arrasto de março a setembro de 2020. Foram coletados 724 espécimes, classificados em 5 famílias, 14 gêneros e 18 espécies. Os resultados gerais revelaram que a maioria da ictiofauna do rio Panjkora contém 8 espécies da família Cyprinidae (56,49%), seguidas por 4 espécies de Nemacheilidae (24,44%), 2 espécies de Channidae (10,63%) e Sisoridae (7,04%) e 1 espécie de Mastacembelidae (1,38%). Entre todas as espécies de peixes, Schizothorax plagiostomus (16,57%) foi altamente predominante, seguida por Carassius auratus (11,87%) e Racoma labiata (9,66%), e elas foram relatadas como altamente abundantes, especialmente nos meses de abril, maio e junho. As espécies menos abundantes foram Glyptothorax punjabensis, Glyptothorax sufii e Mastacembelus armatus, que constituíram 2,48%, 2,20% e 1,38%, respectivamente, do total de peixes amostrados. O índice de diversidade de Simpson (1-D = 0,919), o índice recíproco de Simpson (1/D = 12,3876) e o índice de Shannon (H = 2,68) indicaram que o rio Panjkora contém um grupo bastante rico e diversificado de espécies de peixes. Os microplásticos mais altos foram observados no local 7 em comparação com outra área de estudo. Medidas de conservação devem ser tomadas como prioridade máxima para proteger e conservar o ambiente marinho e o patrimônio natural de novas perdas e extinção e para parar ou minimizar as perdas ocorridas por práticas de pesca irresponsáveis.

[Long Noncoding RNAs MEG3, TUG1, and hsa-miR-21-3p Are Potential Diagnostic Biomarkers for Coronary Artery Disease].

Peripheral blood biomarkers are of particular importance to diagnose certain diseases including coronary artery disease (CAD) due to their non-invasiveness. Investigating the expression of noncoding RNAs (ncRNAs) paves the way to early disease diagnosis, prognosis, and treatment. Consequently, in this research, we aimed to investigate a panel of ncRNAs as potential biomarkers in patients with coronary artery disease. Two different groups have been designed (control and CAD). All participants were subjected to interviews and clinical examinations. Peripheral blood samples were collected, and plasma was extracted. At the same time, target ncRNAs have been selected based on literature review and bioinformatic analysis, and later they underwent investigation using quantitative real-time PCR. The selected panel encompassed the long non-coding RNAs (lncRNAs) MEG3, TUG1, and SRA1, and one related microRNA (miRNA): hsa-miR-21-3p. We observed statistically significant upregulation in MEG3, TUG1, and hsa-miR21-3p in CAD patients compared to control participants (p-value < 0.01). Nevertheless, SRA1 exhibited downregulation with no statistical significance (p-value > 0.05). All ncRNAs under study displayed a significantly strong correlation with disease incidence, age, and smoking. Network construction revealed a strong relationship between MEG3 and TUG1. ROC analysis indicated high potentiality for hsa-miR-21-3p to be a promising biomarker for CAD. Moreover, MEG3 and TUG1 displayed distinguished diagnostic discrimination but less than hsa-miR-21-3p, all of them exhibited strong statistical significance differences between CAD and control groups. Conclusively, this research pinpointed that MEG3, TUG1, and hsa-miR-21-3p are potential biomarkers of CAD incidence and diagnosis.

Immunomodulatory and anti-cancer potential of cloves (Syzygium aromaticum) bud extract and its phytogenic silver nanoparticles.

Clove plant (Syzygium aromaticum) is one of the Myrtaceae family. It's a common flavor in food and the traditional medicine. The study's objective was to ascertain whether the clove bud aqueous extract (CAE) and CAE + nanosilver have any biological effects on immune cells and HT-29 colon cancer cell line. Nanosilver was produced through green synthesis approach using CAE. Produced nanosilver was characterized via electron microscope (scanning, SEM) and ultraviolet-visible spectroscopy. CAE and CAE + nanosilver were examined for their active biomolecules using FTIR analysis, p53 contents using real-time PCR, apoptosis and cell cycle arrest power on HT-29 cancer cell line via flow cytometerty and immunomodulatory potential utilizing MTT assay. Results cleared that a spherical nanosilver with a diameter range of 53 nm was formed by CAE. There were several active biomolecules in CAE and CAE + nanosilver. CAE and CAE + nanosilver increased the p53 protein expression and apoptotic cell number in HT-29 colon cancer cells. CAE and CAE + nanosilver could arrest HT-29 cells at the phase G2/M. CAE and CAE + nanosilver stimulated quiescent and PHA-pre-treated splenic cells at higher concentrations, and CAE suppressed quiescent splenic cell when diluted. In conclusion, the safe edible Syzygium aromaticum plant can be utilized to make anti-tumor agent, essentially for colon tumor. As Syzygium aromaticum plant could stimulate immune cells, it can be used as immune-stimulatory agent that can help fight tumor and tumor development.

New Pyrazole-Clubbed Pyrimidine or Pyrazoline Hybrids as Anti-Methicillin-Resistant Staphylococcus aureus Agents: Design, Synthesis, In Vitro and In Vivo Evaluation, and Molecular Modeling Simulation.

Two hybrid series of pyrazole-clubbed pyrimidines 5a-c and pyrazole-clubbed pyrazoline compounds 6a,b and 7 were designed as attractive scaffolds to be investigated in vitro and in vivo for antibacterial activity against methicillin-resistant Staphylococcus aureus (MRSA) and Pseudomonas aeruginosa. From the results of the in vitro antibacterial screening, compound 5c showed excellent activity (minimal inhibitory concentration, MIC = 521 µM) when compared with that of the reference antibiotic levofloxacin (MIC = 346 µM). The inhibition of the target dihydrofolate reductase (DHFR) enzyme by compounds 4 and 5a-c (IC50 = 5.00 ± 0.23, 4.20 ± 0.20, 4.10 ± 0.19, and 4.00 ± 0.18 µM, respectively) was found to be better than the reference drug trimethoprim (IC50 = 5.54 ± 0.28 µM). Molecular modeling simulation results have justified the order of activity of all the newly synthesized compounds as DHFR enzyme inhibitors, and compound 5c exhibited the best binding profile (-13.6169386 kcal/mol). Hence, the most potent inhibitor of the DHFR enzyme, 5c, was chosen to be evaluated in vivo for its activity in treating MRSA-induced keratitis in rats and that, in turn, significantly (P < 0.0001) reduced infection in rats when compared to MRSA-treated group results.

Novel topoisomerase II/EGFR dual inhibitors: design, synthesis and docking studies of naphtho[2',3':4,5]thiazolo[3,2-a]pyrimidine hybrids as potential anticancer agents with apoptosis inducing activity.

Topoisomerases II are ubiquitous enzymes with significant genotoxic effects in many critical DNA processes. Additionally, epidermal growth factor receptor (EGFR) plays pivotal role in tumour growth and angiogenesis. A novel series of naphtho[2',3':4,5]thiazolo[3,2-a]pyrimidine hybrids have been designed, synthesised and evaluated for their topo IIα/EGFR inhibitory and apoptotic inducer activities. Cytotoxicity of the synthesised hybrids was evaluated against MCF-7, A549 and HCT-116 cell lines. Of the synthesised hybrids, 6i, 6a and 6c experienced superior cytotoxic activity compared to doxorubicin and erlotinib against the tested cancer cells. The molecular mechanism of these hybrids revealed their ability to successfully inhibit topo IIα and EGFR activities in micromolar concentration and may serve as topo II catalytic inhibitor. Moreover, these hybrids significantly arrested cell cycle at G2/M phase together with increased p53, caspae-7, caspase-9 levels and Bax/Bcl-2 ratio. The synthesised hybrids showed efficient binding pattern in molecular docking study and have acceptable drug likeness characters.

Innovative challenge for the inhibition of hepatocellular carcinoma progression by combined targeting of HSP90 and STAT3/HIF-1α signaling.

Hepatocellular carcinoma (HCC) is the third foremost cause of cancer-related deaths. HCC has a very bad prognosis because it is asymptomatic in the early stages, resulting in a late diagnosis, and it is highly resistant to conventional chemotherapy. Such chemotherapies have been proven disappointing because they provide extremely low survival benefits. This study discloses that the STAT3/HIF-1α is an auspicious therapeutic attack site for conceivable repression of HCC development. A site that can be targeted by simultaneous administration of a STAT3 inhibitor in the context of HSP90 inhibition. 17-DMAG binds to HSP90 and constrains its function, resulting in the degradation of HSP90 client proteins HIF-1α and STAT3. Hypoxia recruits STAT3/HIF-1α complex within the VEGF promoter. Additionally, it was acknowledged that STAT3 is an essential mediator of VEGF transcription by direct binding to its promoter. Furthermore, it induces HIF-1α stability and enhances its transcriptional activity. Herein, we revealed that the combination therapy using 17-DMAG and nifuroxazide, a STAT3 inhibitor, repressed the diethylnitrosamine-induced alterations in the structure of the liver. This effect was mediated via decreasing the levels of the HSP90 client proteins HIF-1α and pSTAT3 resulting in the suppression of the STAT3/HIF-1α complex transcriptional activity. To conclude, 17-DMAG/NFXZD combination therapy-induced disruption in the STAT3/HIF-1α loop led to a potential antiangiogenic activity and showed apoptotic potential by inhibiting autophagy and inducing ROS/apoptosis signaling. Additionally, this combination therapy exhibited promising survival prolongation in mice with HCC. Consequently, the use of 17-DMAG/NFXZD renders an inspirational perspective in managing HCC. However, further investigations are compulsory.

Fasudil Ameliorates Methotrexate-Induced Hepatotoxicity by Modulation of Redox-Sensitive Signals.

Methotrexate (MTX) is one of the most widely used cytotoxic chemotherapeutic agents, and it is used in the treatment of different autoimmune disorders. However, the clinical applications of MTX are limited by its hepatic toxicity. Hence, the present study was conducted to evaluate the efficacy of fasudil (Rho-Kinase inhibitor) in the amelioration of MTX hepatotoxicity and the possible underlying mechanisms. Experimentally, 32 male Sprague Dawley rats were used and divided into four groups: control, MTX (20 mg/kg, i.p., single dose), fasudil (10 mg/kg/day i.p.) for one week, and fasudil plus MTX. It was found that MTX significantly induced hepatitis and hepatocellular damage, as shown by abnormal histological findings and liver dysfunction (ALT and AST), with up-regulation of the inflammatory mediators NF-κB-p65 and IL-1ß. Moreover, MTX remarkably disrupted oxidant/antioxidant status, as evidenced by malondialdehyde (MDA) up-regulation associated with the depletion of superoxide dismutase (SOD), catalase, and reduced glutathione (GSH) levels. Moreover, MTX reduced the hepatic expression of B-cell lymphoma 2 (Bcl-2). On the contrary, the i.p. administration of fasudil significantly ameliorated MTX hepatotoxicity by histopathological improvement, restoring oxidant/antioxidant balance, preventing hepatic inflammation, and improving the hepatic anti-apoptotic capability. Furthermore, fasudil hepatic concentration was determined for the first time using the validated RP-HPLC method. In conclusion, the present study revealed that fasudil has a reliable hepatoprotective effect against MTX hepatotoxicity with underlying antioxidant, anti-inflammatory, and anti-apoptotic mechanisms. It also introduced a new method for the determination of fasudil hepatic tissue concentration using the RP-HPLC technique.

Assessment of phytoremediation potential of native plant species naturally growing in a heavy metal-polluted industrial soils.

The present study was carried out in Hayat Abad Industrial Estate located in Peshawar to assess the levels of cadmium (Cd) that were present in the soil as well as the plant parts (Roots and shoots). To evaluate the phytoremediation potential of the plants different factors i.e. Bioconcentration Factor (BCF), Translocation Factor (TF), and Bioaccumulation Coefficient were determined. These plants were grown in their native habitats (BAC). We have analysed, cadmium concentration from soil which are collected from 50 different locations ranged from 11.54 mg/Kg (the lowest) to 89.80 mg/Kg (highest). The maximum concentration (89.80 mg/Kg) of cadmium was found in HIE-ST-16L Marble City and HIE-ST-7 Bryon Pharma (88.51 mg/Kg) while its minimum concentration (12.47 mg/Kg) were detected in the soil of Site (HIE-ST-14L Royal PVC Pipe) and (11.54 mg/Kg) at the site (HIE-ST-11 Aries Pharma). Most plant species showed huge potential for plant based approaches like phyto-extraction and phytoremediation. They also showed the potential for phyto-stabilization as well. Based on the concentration of cadmium the most efficient plants for phytoextraction were Cnicus benedictus, Parthenium hysterophorus, Verbesina encelioides, Conyza canadensis, Xanthium strumarium, Chenopodium album, Amaranthus viridis, Chenopodiastrum murale, Prosopis juliflora, Convolvulus arvensis, Stellaria media, Arenaria serpyllifolia, Cerastium dichotomum, Chrozophora tinctoria, Mirabilis jalapa, Medicago polymorpha, Lathyrus aphaca, Dalbergia sissoo, Melilotus indicus and Anagallis arvensis. The cadmium heavy metals in the examined soil were effectively removed by these plant species. Cerastium dichotomum, and Chenopodium murale were reported to be effective in phyto-stabilizing Cd based on concentrations of selected metals in roots and BCFs, TFs, and BACs values.

Exogenous Ca/Mg quotient reduces the inhibitory effects of PEG induced osmotic stress on Avena sativa L.

Drought is one of the most damaging abiotic stress that hinder plant growth and development. The present study aimed to determine the effects of various Ca/Mg quotients under polyethylene glycol (PEG)-induced osmotic stress on growth, uptake and translocation of Ca and Mg in Avena sativa (L). Plants were grown in nutrient solution supplemented with three different Ca/Mg molar quotients (0.18, 2, and 4). After 30 days plants were exposed to two different PEG (Polyethylene glycol) concentrations (0.6 MPa & 0.2 MPa) for 8 days, and solutions were renewed after 4 days. A solution containing Ca and Mg nutrients has mitigated the negative impact caused via osmotic stress on relative growth rate (RGR), absolute growth rate (AGR), crop growth rate (CGR), leaf area ratio (LAR), Leaf index ratio (LAI), root-shoot ratio (RSR), water use efficiency (WUE) and net assimilation rate (NAR). In addition, it adversely affected germination parameters, including final emergence percentage (FEP), mean germination time (MGT), Timson germination Index (TGI), germination rate index (GRI) and percent field capacity (%FC), of oat (Avena sativa L.). Mg and Ca in shoot and root and Ca translocation factor decreased with increasing Ca in solution, while Mg translocation factor increased with increasing Ca in nutrient solution. In this work, the combined effects of various Ca/Mg quotients and osmotic stress produced by polyethylene glycol (PEG) in different concentrations (0.6 MPa, 0.2 MPa) on the growth and element uptake of Avena sativa L. are examined. As a result, the Ca/Mg Quotient may naturally combat the moderate drought stress experienced by field crops.

Ambroxol, a mucolytic agent, boosts HO-1, suppresses NF-κB, and decreases the susceptibility of the inflamed rat colon to apoptosis: A new treatment option for treating ulcerative colitis.

Ulcerative colitis (UC) is a chronic inflammatory bowel disease of unknown etiology that increases the risk of developing colorectal cancer and imposes a lifelong healthcare burden on millions of patients worldwide. Current treatment strategies are associated with significant risks and have been shown to be fairly effective. Hence, discovering new therapies that have better efficacy and safety profiles than currently exploited therapeutic strategies is challenging. It has been well delineated that NF-κB/Nrf2 crosstalk is a chief player in the interplay between oxidative stress and inflammation. Ambroxol hydrochloride, a mucolytic agent, has shown antioxidant and anti-inflammatory activity in humans and animals and has not yet been examined for the management of UC. Therefore, our approach was to investigate whether ambroxol could be effective to combat UC using the common acetic acid rat model. Interestingly, a high dose of oral ambroxol (200 mg/kg/day) reasonably improved the microscopic and macroscopic features of the injured colon. This was linked to low disease activity and a reduction in the colonic weight/length ratio. In the context of that, ambroxol boosted Nrf2 activity and upregulated HO-1 and catalase to augment the antioxidant defense against oxidative damage. Besides, ambroxol inactivated NF-κB signaling and its consequent target pro-inflammatory mediators, IL-6 and TNF-α. In contrast, IL-10 is upregulated. Consistent with these results, myeloperoxidase activity is suppressed. Moreover, ambroxol decreased the susceptibility of the injured colon to apoptosis. To conclude, our findings highlight the potential application of ambroxol to modify the progression of UC by its anti-inflammatory, antioxidant, and antiapoptotic properties.

Biodiversity, distributions and isolation of microplastics pollution in finfish species in the Panjkora River at Lower and Upper Dir districts of Khyber Pakhtunkhwa province of Pakistan.

The basic aim of this study was aimed to determine the ichthyofaunal diversity of River Panjkora in both upper and lower Dir districts in Khyber Pakhtunkhwa province of Pakistan.Fish samples were collected by using fishnets from March to September 2020. A total of 724 specimens were collected and classified into 5 families, 14 genera, and 18 species. The overall results revealed that most fish fauna of river Panjkora contains 8 species of family Cyprinidae (56.49%) followed by 4 species of Nemacheilidae (24.44%), 2 species of Channidae (10.63%), and Sisoridae (7.04%), and 1 species of Mastacembelidae (1.38%), respectively. Among all kinds of fish species, Schizothorax plagiostomus (16.57%) was highly dominated and followed by Carassius auratus (11.87%) and Racoma labiata (9.66%) and were reported as highly abundant, especially during April, May, and June. The least abundant species were Glyptothorax punjabensis, Glyptothorax sufii, and Mastacembelus armatus, that constituting 2.48%, 2.20%, and 1.38% of the total fish samples. The Overall Simpson's diversity (1-D= 0.919) and Simpson's Reciprocal index values (1/D= 12.3876), and Shannon's index (H= 2.68) were indicating that river Panjkora contains a quite rich and diverse group of fish species. The highest microplastics observed in site 7 compared to other study area. Conservation steps should be taken as a top priority to protect and conserve the marine environment and natural heritage from further loss, extinction and stop or minimize losses incurred through irresponsible fishery practices.

Growth performance and survivability of the Asian seabass Lates calcarifer (Bloch, 1790) reared under hyper-saline, hypo-saline and freshwater environments in a closed aquaculture system.

Salinity is one of the most critical environmental parameters regarding fish physiology, modifying food intake and growth performance in many fish species. The present study has investigated the effects of different salinity levels on growth performance, feeding and survival of Asian seabass Lates calcarifer juveniles. Asian seabass juveniles were reared at 0 (T1), 5 (T2), 22 (T3), 36 (T4), and 42 (T5) ppt salinity. Approximately eight hundred thirty fish individuals with an average weight of 1.24±0.52 g were randomly distributed (166 fish/Tank) in 5 concrete tanks (each tank 30×6×4 ft, volume 19,122 L) for forty days. Juveniles were initially fed 42% crude protein-containing diets at a rate of 6% of their body weight per day. The results showed that salinity level had a significant effect on the weight gain (WG), average daily weight gain (ADWG), specific growth rate (SGR), feed conversion ratio (FCR), survival rate (SR), total biomass and health indices (p<0.05). The highest WG (39.11±1.49 g), ADWG (1.00±0.12 g), SGR (8.74±0.03% d-1) and lowest FCR (0.96±0.20) were observed with T3 treatment, which was significantly higher compared to other treatment groups (p<0.05). Among the health indices, the highest hepatosomatic index and viscerosomatic index were found with T3 treatment, significantly higher than the other groups (p<0.05). No significant differences were found among the treatments in terms of survival rate (p>0.05), but the maximum survival rate (98.89±0.0%) was observed in the T3 and T2 treatments. The maximum level of crude proteins (19.99±1.4%) was found in the whole-body biochemical composition of Asian seabass juveniles in the T3 treatment group. The second-order polynomial regression showed that 20 ppt salinity is optimum for the best growth of Asian seabass. Thus, the present study recommends 20 to 36 ppt salinity for the commercial farming of Asian seabass under a closed aquaculture system.

Nifuroxazide-loaded cubosomes exhibit an advancement in pulmonary delivery and attenuate bleomycin-induced lung fibrosis by regulating the STAT3 and NF-κB signaling: A new challenge for unmet therapeutic needs.

Pulmonary fibrosis (PF) is a chronic progressive disease that portends a very poor prognosis. It has been suggested that STAT3 is a potential target in PF. This study highlights the importance of cubosomes as a drug delivery system in enhancing the bioavailability of nifuroxazide (NXZD), a poorly soluble STAT3 inhibitor. NXZD-loaded cubosomes (NXZD-LC) were in vitro and in vivo evaluated. In vitro, cubosomes presented a poly-angular nanosized particles with a mean size and zeta potential of 223.73 ± 4.73 nm and - 20.93 ± 2.38 mV, respectively. The entrapment efficiency of nifuroxazide was 90.56 ± 4.25%. The in vivo pharmacokinetic study and the lung tissue accumulation of NXZD were performed by liquid chromatography-tandem mass spectrometry after oral administration to rats. The nanoparticles exhibited a two-fold increase and 1.33 times of bioavailability and lung tissue concentration of NXZD compared to NXZD dispersion, respectively. In view of this, NXZD-LC effectively attenuated PF by targeting STAT3 and NF-κB signals. As a result, NXZD-LC showed a potential anti-inflammatory effect as revealed by the significant decrease in MCP-1, ICAM-1, IL-6, and TNF-α and suppressed fibrogenic mediators as indicated by the significant reduction in TGF-ß, TIMP-1, and PDGF-BB in lung tissues. Besides, NXZD-LC improved antioxidant defense mechanisms and decreased LDH and BALF total protein. These effects contributed to decreased collagen deposition. To conclude, cubosomes represent an advantageous pharmaceutical delivery system for enhancing pulmonary delivery of poorly soluble drugs. Additionally, repurposing NXZD as an antifibrotic agent is a promising challenge and new therapeutic approach for unmet therapeutic needs.

Coomassie brilliant blue G-250 dye attenuates bleomycin-induced lung fibrosis by regulating the NF-κB and NLRP3 crosstalk: A novel approach for filling an unmet medical need.

Pulmonary fibrosis (PF) is a life-threatening disorder with a very poor prognosis. Because of the complexity of PF pathological mechanisms, filling such an unmet medical need is challenging. A number of pulmonary diseases have been linked to the activation of NF-κB and the NLRP3 inflammasome. Coomassie brilliant blue G-250 (CBBG) is proved to be a safe highly selective P2×7R antagonist with promising consequent inactivation of NLRP3 inflammasome. This is the first report to investigate the effect of CBBG on the bleomycin-induced lung fibrosis in rats. Our findings revealed that CBBG resulted in a significant improvement in histological features and oxidative status biomarkers of bleomycin-exposed lung tissue. Additionally, CBBG repressed collagen deposition as indicated after the analysis of hydroxyproline, TGF-ß, PDGF-BB, TIMP-1, MMP-9, Col1a1, SMA and ICAM-1. It also exhibited anti-inflammatory potential as revealed by the determination of TNF-α, IL-1ß, IL-18, MCP-1 in the lung tissue. In the bronchoalveolar lavage, the total protein and the LDH activity were substantially reduced. The lung protective effects of CBBG might be attributed on the one hand to the inhibition of NLRP3 inflammasome and on the other hand to the inactivation of NF-κB. Decreased levels of phospho-p65 and its DNA-binding activity as well as the analysis of TLR4 confirmed NF-κB inactivation. Caspase-1 activity is suppressed as a consequence of inhibiting NLRP3 inflammasome assembly. To conclude, CBBG may act as a primary or adjuvant therapy for the management of PF and therefore it may pose an opportunity for a novel approach to an unmet medical need.

Study of some morphometric and meristic characteristics of Alepes vari (Cuvier, 1833) collected from the Arabian coast.

The present study examines the correlations between fifteen morphometric and ten meristic characters and total length (TL) of males, females, and combined sexes of Alepes vari (Cuvier, 1833) collected from Karachi fish harbor, West Wharf of Karachi Coast. Statistical analyses of linear regression relationships show mostly strong correlations (r≥0.70; p<0.05) between total length (TL) and most morphometric characters in males, females, and combined sexes, except the height of pectoral-fin (PFH), and pelvic-fin base length (PelFL); whereas, meristic characters were found to be constant and indicate weak or negative type correlations (r≤0.50; p>0.05) with total length (TL). Hence, according to our present results, there is a direct relationship between the total length of fish and all morphometric characters, which were found to be the best indicators of positive allometric pattern growth in fish. Moreover, analysis of the 2-sample t-test revealed (t-test; p>0.05) that no sexual dimorphism was reported in Alepes vari. Thus, our present study could be valuable in systematic classification, sexual dimorphism, and management of this species on the Karachi coast.