Withaferin A alleviates inflammation in animal models of arthritis by inhibiting the NF-κB pathway and cytokine release.

Withaferin A, a steroid lactone from Withania somnifera, exhibits anti-inflammatory, immunomodulatory, and antioxidant properties. This study investigated the effects of withaferin A on collagen-induced arthritis (CIA) rats, focusing on NF-κB p65 regulation and cytokine release. Withaferin A (50 mg/kg b.wt., orally) or methotrexate (0.25 mg/kg b.wt., i.p., as a reference drug) was given to CIA rats daily for 20 days postarthritis induction. Joints were removed from nonarthritic and arthritic rats to assess the levels of NO, MPO, interleukin (IL)-1ß, IL-6, IL-10, TNF-α, COX-2, and NF-κB via ELISA. Furthermore, the mRNA expression of IL-1ß, IL-10, TNF-α, COX-2, iNOS, and NF-κB was also assessed through qPCR. Treatment with withaferin A significantly inhibited the levels of inflammatory cytokines and the transcription factor NF-κB; suppressed the expression of IL-1ß, IL-10, TNF-α, COX-2, iNOS, and NF-κB in the joint tissue of CIA rats; and reduced cartilage and bone destruction, as shown by H&E staining. To confirm the results obtained from biochemical and molecular studies and to determine the molecular target of withaferin A, we performed a molecular simulation of the potential targets of withaferin A, which identified the NF-κB pathway as its target. These results suggested that withaferin A effectively attenuated rheumatoid arthritis progression by inhibiting the activation of the NF-κB pathway and the downstream secretion of inflammatory cytokines.

Novel Insights in the Hypertension Treatment & Type 2 Diabetics Induced by Angiotensin Receptor Blockers: MD Simulation Studies & Molecular Docking of Some Promising Natural Therapies.

Angiotensin receptor blockers (ARBs) are commonly used to treat hypertension that target the hormonal system (renin-angiotensin system (RAS)), which regulates various physiological functions in the body. ARBs work by blocking the binding of angiotensin II to its receptor, thereby preventing a rise in blood pressure. These drugs not only normalize the overactivation of RAS but also provide protective effects against cardiovascular, renal, and type 2 diabetic patients. Inappropriate RAS activity has been linked to insulin resistance of type 2 diabetes. Olmesartan, as an ARB, was found to have a beneficial role in reducing postprandial glucose levels in type 2 diabetes. However, ARBs can cause side effects, prompting a search for new compounds that have fewer adverse effects. This study explores the potential of natural metabolites, specifically eugenol, gallic acid, myricetin, p-cymene, quercetin, and kaempferol, as ARB inhibitors compared to the current standard, olmesartan. Using in silico studies, the binding affinity of these natural substances to the ARB receptor was evaluated. The results showed that myricetin and kaempferol had affinities higher than those of olmesartan, suggesting that they could serve as promising ARB inhibitors for hypertension treatment. These natural compounds could provide an alternative approach to conventional antihypertensive drugs, which may have fewer side effects. However, more research is needed to validate the efficacy and safety of these natural compounds as antihypertensive drugs. Further in vitro and in vivo studies are needed to confirm their effectiveness and safety. This study provides a promising starting point for future investigations into the potential of natural metabolites as alternative treatments for hypertension. The findings also highlight the importance of exploring natural alternative treatments for hypertension and the protective effects of ARBs on early stage type-2 diabetics.

The Rapidly Changing Patterns in Bacterial Co-Infections Reveal Peaks in Limited Gram Negatives during COVID-19 and Their Sharp Drop Post-Vaccination, Implying Potential Evolution of Co-Protection during Vaccine-Virus-Bacterial Interplay.

SARS-CoV-2 has caused the most devastating pandemic of all time in recent human history. However, there is a serious paucity of high-quality data on aggravating factors and mechanisms of co-infection. This study aimed to identify the trending patterns of bacterial co-infections and types and associated outcomes in three phases of the pandemic. Using quality hospital data, we have investigated the SARS-CoV-2 fatality rates, profiles, and types of bacterial co-infections before, during, and after COVID-19 vaccination. Out of 389 isolates used in different aspects, 298 were examined before and during the pandemic (n = 149 before, n = 149 during). In this group, death rates were 32% during compared to only 7.4% before the pandemic with significant association (p-value = 0.000000075). However, the death rate was 34% in co-infected (n = 170) compared to non-co-infected patients (n = 128), indicating a highly significant value (p-value = 0.00000000000088). However, analysis of patients without other serious respiratory problems (n = 28) indicated that among the remaining 270 patients, death occurred in 30% of co-infected patients (n = 150) and only 0.8% of non-co-infected (n = 120) with a high significant p-value = 0.00000000076. The trending patterns of co-infections before, during, and after vaccination showed a significant decline in Staphylococcus aureus with concomitant peaks in Gram negatives n = 149 before/n = 149 during, including Klebsiella pneumonian = 11/49 before/during, E. coli n = 10/24, A. baumannii n = 8/25, Ps. aeruginosa n = 5/16, and S. aureus 13/1. Nevertheless, in the post-vaccination phase (n = 91), gender-specific co-infections were examined for potential differences in susceptibility. Methicillin-resistant S. aureus dominated both genders followed by E. coli in males and females, with the latter gender showing higher rates of isolations in both species. Klebsiella pneumoniae declined to third place in male patients. The drastic decline in K. pneumoniae and Gram negatives post-vaccination strongly implied a potential co-protection in vaccines. Future analysis would gain more insights into molecular mimicry.

Praziquantel efficacy in mice infected with PZQ non-susceptible S. mansoni isolate treated with artemether: parasitological, biochemical and immunohistochemical assessment.

Based on the fact that artemether (ART) affects immature schistosomes and that the effect of praziquantel (PZQ) mainly targets mature schistosomes, this work investigates the possible enhanced efficacy of PZQ in combination with ART in mice harboring a PZQ non-susceptible Schistosoma mansoni isolate. Associated schistosomal, inflammatory, hepatic histopathological changes have been investigated by examining the tissue markers expressing apoptosis using FAS (CD95), anti-apoptosis (Bcl2) and angiogenesis [vascular endothelial growth factor (VEGF)]. A batch of Swiss albino mice infected with a PZQ non-susceptible (EE10) S. mansoni isolate was divided into 12 groups. Animals of the first group were left without treatment as infected controls, while groups 2-6 received PZQ in increasing doses. The animals of group 7 received ART in double doses. Those comprising groups 8-12 received combined therapy of PZQ and ART in the same doses and at the same timings postinfection (PI) as those belonging to groups 2-6. Parasitological parameters, liver function, and histopathological and immunohistochemical studies of FAS, Bcl2 and VEGF antibodies were assessed. Combined administration of ART and PZQ reduced the ED(50) (the dose at which the worm burden was decreased by 50%) of PZQ. Typical granulomas were not seen in animals treated with ART alone and combined with PZQ, with least expression of FAS and VEGF and increased expression of Bcl2. The minimal histopathological changes recorded in mice treated with both ART and PZQ could be related to a synergistic/additive effect of ART, markedly reducing the intensity of infection. Improved liver function tests support the less severe histopathological changes under the influence of this treatment protocol. This study encourages human trials especially in areas where malaria is not endemic, and differing combination doses should be investigated in view of the antagonistic effect noticed with some dose regimens.

Immunohistopathological and biochemical changes in Schistosoma mansoni-infected mice treated with artemether.

OBJECTIVE: To investigate immune mechanisms possibly involved in the amelioration of histopathological changes in livers of Schistosoma mansoni-infected mice treated with artemether (ART), including liver functions and apoptotic changes. METHODS: Male CD-1 Swiss albino mice were infected with Schistosoma mansoni and treated with praziquantel (PZQ) 6 weeks post-infection (PI) (500 mg/kg/day x2) and/or ART in double dose (each of 400 mg/kg) 4 and 6 weeks PI. Parasitological parameters, liver functions and histopathological changes including T-lymphocyte profile and apoptotic changes were assessed. RESULTS: Eight weeks PI, although the reduction in worm burden in mice treated with ART plus PZQ was comparable to that in PZQ-treated mice, yet there was complete absence of eggs and typical granulomas. The ratio of T-helper/cytotoxic cells was in favor of T-helper in infected control and in mice treated with both drugs. This ratio was 0.9:1 and 0.7:1 in PZQ and ART-treated groups, respectively, with moderate apoptotic changes in the latter. All biochemical parameters expressing liver function were improved with all treatment regimens. CONCLUSIONS: Administration of ART in addition to PZQ resulted in absence of eggs and typical granulomas with less apoptotic changes than in ART-treated mice. Improved liver functions with higher apoptosis in ART-treated mice may suggest enhanced necrotic cell death/regenerative changes.

Antipyrine clearance in comparison to conventional liver function tests in hepatitis C virus patients.

In this study, 15 healthy volunteers and 96 patients with hepatitis C virus, classified according to Child-Pugh into 36 Child-A, 31 Child-B and 29 Child-C, were examined. All subjects ingested 600 mg antipyrine in the form of hard gelatinous capsules after overnight fasting. One milliliter of saliva was collected at 4 and 24 h after ingestion of antipyrine and analyzed using high-performance liquid chromatography. Blood samples were collected from all subjects for examination, using conventional liver function tests. The pharmacokinetic variables for antipyrine were determined using the two concentration time points selected. A cut-off value of 0.34 ml/min/kg was used to distinguish between cirrhotic and non-cirrhotic patients. Alanine aminotransferase, aspartate aminotransferase and gamma-glutamyl transferase values were significantly higher with significantly lower antipyrine clearance in Child-A, B, and C patients than in normal volunteers. The total protein concentration was significantly lower in Child-B and C patients. Moreover, AST was significantly higher in Child-C patients and antipyrine clearance was lower in Child-B and C patients than in Child-A patients. Antipyrine clearance showed a significant negative correlation with Child-Pugh scores, total protein, the international normalization ratio of prothrombin time and globulin, and a positive correlation with albumin and albumin-to-globulin ratio. Unlike most of the conventional liver function tests, antipyrine clearance, which represents the intrinsic clearance capacity of the liver, measured using saliva, proved to be a sensitive marker of liver function. It was significantly impaired in the Child-Pugh group A patients with the least hepatic impairment. The international normalization ratio of prothrombin time was just as informative as antipyrine clearance in identifying minimal hepatic impairment.

Effect of adamantylamide dipeptide as adjuvant therapy to praziquantel in mice infected with different S. mansoni isolates.

This work investigated the possible use of AdDP as adjuvant therapy to praziquantel (PZQ) in mice infected with PZQ-insusceptible Schistosoma mansoni isolate in a trial to increase the susceptibility of this isolate to the drug. Two batches of C57 BL/6 mice were infected with PZQ-susceptible and -insusceptible S. mansoni isolates, and each batch was divided into five groups. Seven weeks postinfection, the experimental group received AdDP (5 mg/kg) in addition to PZQ in reduced dose (3x100 mg/kg). Three of the remaining four groups were treated controls; they received AdDP, PZQ in reduced dose and in full dose (2x500 mg/kg), and the fourth group was infected untreated. In mice infected with PZQ-susceptible or -insusceptible S. mansoni isolate, praziquantel alone, and in addition to AdDP, reduced worm and egg loads and increased percentage dead eggs. Also, they improved the histopathological changes (reduction in granuloma diameter, percentage fibrotic area with increased percentage degenerated eggs). Inducible nitric oxide synthase (iNOS), nitric oxide (NO) in culture of peritoneal macrophages, and number of CD68-positive cells were decreased with improved alanine amino transaminase. In mice receiving combined therapy AdDP+PZQ, the antischistosomal efficacy and the reductions in the inflammatory granulomatous reactions, NO in cultured peritoneal macrophages, percentage fibrotic areas recorded, were comparable to that in mice receiving full dose of PZQ, with significantly higher reduction in CD68 cells denoting enhanced antischistosomal efficacy and healing of the inflammatory reactions in the liver.

Effect of combined chemotherapy and anti-inflammatory drugs on murine schistosomiasis.

Hepatic granulomas resulting from Schistosoma mansoni infection contain high levels of prostaglandins (PG) and leukotrienes. The present experimental study was conducted to show the effect of murine S. mansoni infection on PGE2 level in the liver granuloma homogenate and to explore the possibility of using non-steroidal anti-inflammatory drugs (NSAIDs) namely, ibuprofen (CAS 15687-27-1) and naproxen (CAS 22204-53-1), either alone or in combination with praziquantel (CAS 55268-74-1) to induce regression of hepatic morbidity or to ameliorate the biochemical and histopathological consequences and intensity of infection. Infection with S. mansoni increased the level of alanine amino-transaminase (ALT), gamma glutamyl transferase (GGT), PGE2, hepatic collagen deposition, antibody titre and circulating schistosomal antigen. The last parameter was reduced significantly by progression of infection in the 11th and 16th weeks. Treatment with praziquantel was found to reduce the number of worms (97%), with complete absence of immature ova and increase in the number of dead ova. Also it reduced all the elevated parameters except PGE2. NSAIDs were found to reduce significantly the level of PGE2 at 9 weeks but not at 11 and 16 weeks post-infection. ALT and GGT were not significantly decreased compared to their corresponding controls. Treatment with ibuprofen or naproxen either alone or in combination with praziquantel or praziquantel alone reduced significantly the granuloma diameters. Collagen deposition and percentage of fibrotic area were significantly decreased compared to infected control but the antibody titre or circulating antigen levels were not affected. Combined therapy of praziquantel with ibuprofen or naproxen improved most of the parameters estimated and maintained the reducing effect on PGE2 at 11 and 16 weeks post-infection. So it can be concluded that in S. mansoni infection treatment with ibuprofen or naproxen is not preferable without treatment of schistosomiasis by using praziquantel.