Topical nano-delivery of 5-fluorouracil: Preparation and characterization of water-in-oil nanoemulsion

Purpose: To prepare and characterize a water-in-oil nanoemulsion of 5-fluorouracil (5FU) for enhanced skin penetration.Methods: Nanoemulsions of 5FU were prepared using Capyrol (propylene glycol monocaprylate). Transcutol (highly purified diethylene glycol monoethyl ether) and polyethylene glycol (PEG) 400 as oil, surfactant and co-surfactant, respectively. The optimized formulations were subjected to heating - cooling cycling, centrifugation and freeze - thaw cycling to assess their stability. Particle size distribution and zeta potential of the nanoemulsions were evaluated. Furthermore, in vitro and in vivo skin permeation studies were carried out on the formulations in a rat model. Skin irritation studies were also performed on rats to assess the irritation potential of the formulations. The 1 % w/v of Carbopol 934 gel loaded with 1 % 5FU was used as control (FU gel).Results: The results showed that the mean droplet size of the nanoemulsions was ~100 nm with a zeta potential of ± 15. Significant increase in permeability was also observed for the nanoemulsion formulations compared with control. The steady-state flux (Jss), enhancement ratio and permeability coefficient (Kp) for optimized nanoemulsion formulation were significantly (p < 0.05) higher than those of the conventional gel (control). Both in vitro and in vivo skin retention results indicate higher drug release from the nanoemulsion (292.45 μg/cm2) than for control (121.42 μg/cm2). Mean irritation index for the nanoemulsion was significantly lower than for control.Conclusion: The results suggest that a water-in-oil nanoemulsion is safe and can potentially be used to promote skin penetration of 5FU following topical application on the skin for the treatment of some skin diseases.Keywords: Nanoemulsion, Controlled release, 5-Fluorouracil, Skin penetration, Skin irritatio

Effects of nanosuspension and inclusion complex techniques on the in vitro protease inhibitory activity of naproxen

This study investigated the effects of nanosuspension and inclusion complex techniques on in vitro trypsin inhibitory activity of naproxen—a member of the propionic acid derivatives, which are a group of antipyretic, analgesic, and non-steroidal anti-inflammatory drugs. Nanosuspension and inclusion complex techniques were used to increase the solubility and anti-inflammatory efficacy of naproxen. The evaporative precipitation into aqueous solution (EPAS) technique and the kneading methods were used to prepare the nanosuspension and inclusion complex of naproxen, respectively. We also used an in vitro protease inhibitory assay to investigate the anti-inflammatory effect of modified naproxen formulations. Physiochemical properties of modified naproxen formulations were analyzed using UV, IR spectra, and solubility studies. Beta-cyclodextrin inclusion complex of naproxen was found to have a lower percentage of antitryptic activity than a pure nanosuspension of naproxen did. In conclusion, nanosuspension of naproxen has a greater anti-inflammatory effect than the other two tested formulations. This is because the nanosuspension formulation reduces the particle size of naproxen. Based on these results, the antitryptic activity of naproxen nanosuspension was noteworthy; therefore, this formulation can be used for the management of inflammatory disorders.O objetivo do presente estudo foi investigar a atividade anti-inflamatória in vitro de nanossuspensões e do complexo de inclusão contendo naproxeno. Esse fármaco é derivado de ácido propiônico, com ação analgésica, antipirética e antiinflamatória. A obtenção dessas formulações teve por finalidade o aumento da solubilidade e da atividade anti-inflamatória do fármaco. Os métodos por precipitação em solução aquosa por evaporação e por empastagem foram modificados para a obtenção da nanossuspensão e do complexo de inclusão, respectivamente. Para a avaliação da atividade anti-inflamatória das formulações utilizou-se ensaio in vitro modificado de inibição de tripsina. As propriedades físico-químicas das formulações propostas foram determinadas utilizando espectroscopia UV e de infravermelho, além de estudos de solubilidade. O complexo de inclusão de naproxeno apresentou menor atividade antitripsina, quando comparado ao composto livre e à nanossuspensão. Em conclusão, entre as formulações avaliadas, a nanossuspensão de naproxeno apresentou maior efeito anti-inflamatório. Esse efeito foi devido à redução da dimensão das partículas de naproxeno para a escala nanométrica. Com base nos resultados obtidos, a atividade da nanossuspensão de naproxeno foi notável. Dessa forma, essa formulação apresenta potencial para o tratamento de distúrbios inflamatórios

Modulation of glucose transporter proteins by polyphenolic extract of Ichnocarpus frutescens (L.) W. T. Aiton in experimental type 2 diabetic rats

172-180Traditionally, in India, the decoction of Black creeper, Ichnocarpus frutescens (L.) W. T. Aiton leaves is used to treatment diabetes mellitus. However, its molecular mechanisms of antihyperglycemic effects have not been completely studied. Due to the potential antidiabetic effect of I. frutescens, we hypothesized that the polyphonic extract might add to glucose uptake through improvement in the expression of genes of the glucose transporter (GLUT) family messenger RNA (mRNA) in the liver and adipose tissues. Experimentally, diabetes mellitus was induced in Wistar rats through i.p. injection of freshly prepared solution of streptozotocin (45 mg/kg). This was done 15 minutes after the administration of nicotinamide (120 mg/kg, ip). Serum level of insulin and C-peptide were analyzed using standard methods. Glucose metabolism by the hepatocytes and adipocytes were also analyzed by quantitative RT-PCR mRNA expression levels of phosphoenolpyruvate carboxykinase 1 (PCK1), GLUT2 in the hepatocytes, and GLUT4 in the adipocytes. The hemidiaphragm were also isolated and processed to study in-vitro peripheral glucose utilization. Results of the present investigation suggest that STZ-NA induced diabetes is associated with hyperglycemia, altered levels of PCK1 and glucose transporters gene expression as well as decreased levels of insulin and C-peptide. On the other hand, the outcome of the daily oral administration of PPE to STZ-NA induced diabetic rats at different doses (150 and 300 mg/kg bodywt.) for 30 days supports our hypothesis by showing significant improvement of insulin levels, C-peptide level, downregulation of PCK1 and upregulation of GLUT (2, 4) mRNA expression levels when compared to those of diabetic rats. The administration of PPE had also increased the uptake of glucose by rat hemidiaphragm significantly. Findings from this study demonstrate that PPE enhances peripheral glucose uptake through glycogenesis pathway, mediated by upregulation of GLUT2 and GLUT4, and downregulation of PCK1. Our study suggests that the leaf of I. frutescens is a rich source of polyphenolic compounds, including those with an insulin-sensitizing function that may have the potential for treating or managing diabetes or insulin resistance

Hepatoprotective effect of combined extracts of Andrographis paniculata, Boerhaviadiffusa, Eclipta alba and Picrorhiza kurroa on carbon tetrachloride andparacetamol-induced hepatotoxicity in rats

Extracts obtained from Andrographis paniculata, Boerhavia diffusa, Eclipta alba and Picrorhiza kurroa were mixed in an equal ratio and the combined extract was evaluated for the hepatoprotective activity against carbon tetrachloride and paracetamol induced hepatotoxicity in rats. The hepatotoxicants increased the levels of transaminases, total bilirubin, triglycerides and decreased the level of albumin in the serum of rat. Liver homogenate of the rats showed an increase in the lipid peroxidation level and reduction in the levels of glutathione. The rats administered with the combined extract reversed the biochemical parameters reflecting the liver function. The combined extract showed significant (P < 0.05) dose dependent hepatoprotective activity against the toxicants as it was evident from the values of biochemical parameters when compared to silymarin administered animals. The results showed that the recovery exhibited by the combined extract at 400 mg/kg dose level was highest among other doses tested and was well comparable to silymarin treated rats

Anti-Urolithiatic Activity of Melia Azedarach Linn Leaf Extract in Ethylene Glycol-Induced Urolithiasis in Male Albino Rats

Purpose: To investigate the anti-urolithiatic activity of the aqueous and alcoholic extracts of Melia azedarach Linn leaves in calcium oxalate urolithiasis in male albino rats.Methods: The effect of oral administration of aqueous and ethanol extracts of Melia azedarach Linn leaves on calcium oxalate urolithiasis has been investigated. Lithiasis was induced by oral adminstration of ethylene glycol (0.75 %v/v) in male albino rats for 28 days. Each of the extract (250 mg/kg) was administered orally day 0 as a prophylactic regimen and from day 15 as a curative regimen. Regular administration of ethylene glycol caused hyperoxaluria in ethylene glycol-fed animals, leading to increased renal retention and excretion of oxalate, calcium and phosphate. Histopathological study, urine microscopy, serum analysis and biochemical analysis of kidney homogenate were performed.Results: Oxalate and calcium excretion in urine increased (p < 0.01) to 3.68 ± 0.01 and 4.5 ± 0.01 mg/24 h, respectively, in lithiatic control animals compared to (0.37 ± 0.01 and 1.27 ± 0.12 mg/24 h) for the normal control group. Treatment with aqueous or ethanol extract (250 mg/kg, p.o.) significantly (p <0.01) reduced the elevated levels of calcium, oxalate and phosphate excretion in urine to 0.79 ± 0.01 and 1.09 ± 0.04 mg/24 h, respectively. Following treatment with the ethanol extract (250mg/kg), serum creatinine excretion was restored from 0.95 ± 0.01 mg/24 h to the normal level of 0.87 ± 0.01 mg/24 h. The results were comparable to those of the standard drug, allopurinol (50 mg/kg p.o.).Histopathological data for the kidney supported the foregoing results.Conclusions: The results demonstrate that the aqueous and ethanol extracts of Melia azedarach Linn leaves have potent antiurolithiatic activity against ethylene glycol-induced calcium oxalate urolithiasis in male albino rats.Keywords: Melia azedarach, Antiurolithiatic, Ethylene glycol, Urolithiasis, Excretion, Kidne

INVESTIGATION ON ANTIDIARRHOEAL ACTIVITY OF ARISTOLOCHIA INDICA LINN. ROOT EXTRACTS IN MICE

Background: The present study aimed at investigating the effect of ethanolic extract (EtAI), and aqueous extract (AqAI) of Aristolochia indica Linn roots on castor oil-induced diarrhoea and study on small intestinal transit. Phytochemical analysis of extracts was performed as per standard procedure. Materials and Methods: The oral toxicity study using Swiss albino mice was performed in accordance with OECD guidelines. The EtAI and AqAI extracts of Aristolochia indica Linn were studied for antidiarrhoeal property using castor oil-induced diarrhoeal model and charcoal-induced gastrointestinal motility test in Swiss albino mice. Results: Among the tested doses of 200 and 400 mg/kg body weight, the extracts reduced the frequency and severity of diarrhoea in test animals throughout the study period. At the same doses, the extract delayed the intestinal transit of charcoal meal in test animals as compared to the control and the results were statistically significant. Conclusion: Experimental findings showed that ethanol extract of Aristolochia indica Linn root possess significant antidiarrheal activity and may be a potent source of anti-diarrhoeal drug in future

Genome editing and cancer: How far has research moved forward on CRISPR/Cas9?

Financiado para publicación en acceso aberto: Universidade de Vigo/CISUGCancer accounted for almost ten million deaths worldwide in 2020. Metastasis, characterized by cancer cell invasion to other parts of the body, is the main cause of cancer morbidity and mortality. Therefore, understanding the molecular mechanisms of tumor formation and discovery of potential drug targets are of great importance. Gene editing techniques can be used to find novel drug targets and study molecular mechanisms. In this review, we describe how popular gene-editing methods such as CRISPR/Cas9, TALEN and ZFNs work, and, by comparing them, we demonstrate that CRISPR/Cas9 has superior efficiency and precision. We further provide an overview of the recent applications of CRISPR/Cas9 to cancer research, focusing on the most common cancers such as breast cancer, lung cancer, colorectal cancer, and prostate cancer. We describe how these applications will shape future research and treatment of cancer, and propose new ways to overcome current challengesKing Khalid University in Abha, Saudi Arabia | Ref. RGP: 52/2/144

Codon Usage Is Influenced by Compositional Constraints in Genes Associated With Dementia

Dementia is a clinical syndrome characterized by progressive cognitive decline, and the symptoms could be gradual, persistent, and progressive. In the present study, we investigated 47 genes that have been linked to dementia. Compositional, selectional, and mutational forces were seen to be involved. The influence of these two compositional constraints on codon usage bias (CUB) was positive for nucleotide A and negative for GC. Nucleotide A also experienced the highest mutational force, and GC-ending codons were preferred over AT-ending codons. A high bias towards GC-ending codons enhanced the gene expression level, evidenced by the positive association between CAI and GC-ending codons. The unusual behavior of TTG codon showing an inverse relationship with GC-ending codon and negative influence of gene expression, a behavior contrary to all other GC-ending codons, shows operative selectional force. Furthermore, parity analysis, higher translational selection value, preference of GC-ending codons over AT-ending codons, and the association of gene length with gene expression refer to the dominant role of selection pressure with compositional constraint and mutational force shaping codon usage

Therapeutic implications of current Janus kinase inhibitors as anti-COVID agents: A review

Severe cases of COVID-19 are characterized by hyperinflammation induced by cytokine storm, ARDS leading to multiorgan failure and death. JAK-STAT signaling has been implicated in immunopathogenesis of COVID-19 infection under different stages such as viral entry, escaping innate immunity, replication, and subsequent inflammatory processes. Prompted by this fact and prior utilization as an immunomodulatory agent for several autoimmune, allergic, and inflammatory conditions, Jakinibs have been recognized as validated small molecules targeting the rapid release of proinflammatory cytokines, primarily IL-6, and GM-CSF. Various clinical trials are under investigation to evaluate Jakinibs as potential candidates for treating COVID-19. Till date, there is only one small molecule Jakinib known as baricitinib has received FDA-approval as a standalone immunomodulatory agent in treating critical COVID-19 patients. Though various meta-analyses have confirmed and validated the safety and efficacy of Jakinibs, further studies are required to understand the elaborated pathogenesis of COVID-19, duration of Jakinib treatment, and assess the combination therapeutic strategies. In this review, we highlighted JAK-STAT signalling in the pathogenesis of COVID-19 and clinically approved Jakinibs. Moreover, this review described substantially the promising use of Jakinibs and discussed their limitations in the context of COVID-19 therapy. Hence, this review article provides a concise, yet significant insight into the therapeutic implications of Jakinibs as potential anti-COVID agents which opens up a new horizon in the treatment of COVID-19, effectively