ADRENAL HISTOPLASMOSIS: AN EASTERN INDIAN PERSPECTIVE.

Context: The clinical presentation of histoplasmosis is varied. Due to its propensity for adrenal involvement, histoplasmosis is an important differential diagnosis in any patient presenting with adrenal mass, bilateral in particular. Objective: Data on clinical presentation, pattern of adrenal involvement, radiological appearance and long-term follow-up of adrenal histoplasmosis are relatively sparse; hence we looked at it. Design: This record based single-centre retrospective study was conducted in one of the tertiary care hospitals, situated in eastern India catering the Gangetic delta. Subjects and methods: Data on demographic characters, presenting manifestations, biochemical & hormonal parameters and radiological appearance of confirmed adrenal histoplasmosis cases (n=9), admitted between 2015-2019 have been retrieved. The treatment outcome and condition of patients after 1-4 years of follow-up has also been discussed. Results: Four out of the nine (44.4%) patients had predisposing immunocompromised conditions in the form of diabetes and/or chronic alcoholism while rest were immunocompetent. Seven out of nine patients (77.8 %) had signs and symptoms suggestive of adrenal insufficiency, while two (22.2%) presented with only pyrexia of unknown origin. All of them had bilateral adrenal mass, though the radiologically appearances were different. All patients received anti-fungal agents with/without hydrocortisone and/or fludrocortisone. One patient died (11.1%), while majority responded favourably to treatment. Adrenocortical function did not recover completely. Conclusions: The possibility of adrenal histoplasmosis should always be considered in patients presenting with bilateral adrenal mass, irrespective of adrenal morphology. Treatment is effective, but many of them require supplemental hydrocortisone for quite a long period, if not lifelong. Mineralocorticoid deficiency, however, is not permanent.

A novel synergistic device for joint inflammation - efficacy on ankle sprain cases.

Joint inflammation results from soft-tissue injuries and cartilage damage. PRICE is the standard treatment approach for acute soft tissue injuries like ankle sprain. Electrical stimulation, application of orthotic braces, etc. is also effective for this. In a synergistic device all these components are combined and applied simultaneously. This device was developed for joint inflammation and tested for grade I & II acute ankle joint sprain. To test a synergistic - semirigid device, combining PRICE & electrical stimulation for acute ankle sprains of grades I & II for pain, range of motion and swelling is a case series was the objective. Device was developed using novel concept of synergistic applications of PRICE with electrical stimulation. The joint contour of ankle and specific biomechanical bony surface landmarks were considered. Ethical approval was taken from NTCC committee, AIPT. Recordings were taken from eight patients of acute ankle sprain with - in two days of injury, after getting ethical approval. Elevation to the ankle was provided by keeping the part over the pillow and data was recorded with the help of: 1.VAS scale for pain; 2. Measuring tape; 3. Goniometry. t-test was used to find out the significant difference pre and post the application of device. There was a significant reduction in pain (p = 0.006), edema (p = 0.011), dorsi-flexion (p = 0.015), and plantar flexion (p =0.008). The synegistic device has been effective for acute ankle inflammation - grade I & II ankle sprains in 5 sessions; sufficient for the return of function.

Boswellic acid inhibits inflammatory angiogenesis in a murine sponge model.

The aim of the present study was to investigate the effects of boswellic acid (BA) on key components of inflammatory angiogenesis in the murine cannulated sponge implant angiogenesis model. Polyester-polyurethane sponges, used as a framework for fibrovascular tissue growth, were implanted in Swiss albino mice and BA (12.5 or 25mg/kg/day) was given through installed cannulas for nine days. The implants collected at day 9 post-implantation were processed for the assessment of hemoglobin (Hb). Relevant levels of inflammatory, angiogenic and fibrogenic cytokines were also determined. BA treatment resulted in significant decrease in sponge vascularization (Hb content) and in vascular endothelial growth factor (VEGF) and transforming growth factor (TGF-ß1) at both doses. Further, BA decreased expression of VEGF and CD31 and reduced % microvessel density (MVD) in sponge implants. A regulatory function of BA on multiple parameters of the main components of inflammatory angiogenesis has been revealed giving an insight into the potential therapeutic use underlying the actions of BA.

Anti-ovulatory activity of H2 receptor blockers in albino rabbits--a preliminary study.

OBJECTIVE: To evaluate the anti-ovulatory activity of H(2) receptor blockers (ranitidine, famotidine and roxatidine) in albino rabbits considering the role of histamine in ovulation. METHOD: The drugs were orally administered once daily for three days to adult female rabbits weighing between 1.3-2.0 kg (four groups of three animals). The control group received the 1% weight/volume gum acacia suspension. Thirty minutes after the administration of the last dose, a freshly prepared 0.4 % solution of cupric acetate was administered to each animal intravenously via the marginal ear vein (4 mg/kg body weight) to induce ovulation. To assess ovulation, laparotomy was carried out 48 h after cupric acetate injection. The ovaries were exposed, bleeding points on each ovary were counted, and the ovaries and uteri were subjected to histopathological evaluation. RESULTS: Based on the number of bleeding points (ovulation sites) observed on the ovary, H(2) blockers showed varying degrees of anti-ovulatory activity. Roxatidine exerted the most pronounced activity. Histopathological observations of uterus and ovary confirmed the aforementioned observations. CONCLUSION: H(2) receptor blockers appeared to inhibit the cupric acetate-induced ovulation in albino rabbits. Our results seem to confirm the role of histamine in ovulation reported by other authors.

Standardization of Berberis aristata DC and Nigella sativa L. Using HPTLC and GCMS and Their Antineoplasia Activity in 7,12-Dimethylbenz[a]anthracene-Induced Mouse Models.

Berberis aristata DC and Nigella sativa L. are officially listed in various Indian Pharmacopoeia and AYUSH official documents. Prescribed for different ailments for proven medicinal activities, they thus became part of polyherbal medications. With reverse pharmacology and scientific validation, more than 30 patents are filed on different formulations of B. aristata and granted. Nigella sativa L. has been broadly studied for its therapeutic potential and wide range of activities against cardiovascular, diabetic, cancer, and life style disorders. Thus, this study is aimed at standardizing B. aristata and N. sativa and their antineoplasia activity in 7, 12-dimethylbenz[a]anthracene (DMBA)-induced mouse models. Molecular docking was done using the Schrodinger program Maestro 9.0. Herbal extracts and essential oil (B. aristata and N. sativa) were standardized and quantified using high-performance thin-layer chromatography (HPTLC) (CAMAG) and gas chromatography-mass spectrometry (GCMS) (Agilent 2010GC System) with validated methods. DMBA was administered orally once a week (1mg/200 µL) to each animal except the normal control. Hematology, histopathology, and immunoassays were performed, and data were analyzed and depicted with GraphPad and SPSS. In molecular docking, thymoquinone showed the highest docking score (9.519, 9.211, and 9.042, respectively) in the active site pockets of IL6 (PDB ID: 4CNI and 5FCU), TNF (PDB ID: 2AZ5), and VEGF (PDB ID: 4KZN). Out of all four target sites, thymoquinone and berberine showed good binding affinity with IL6 (PDB ID: 4CNI) compared to α- and ß-pinenes. HPTLC analysis of the hydroalcoholic extract showed the presence of berberine both qualitatively and quantitatively (5.4% berberine), and thymoquinone detected 0.17% in the N. sativa extract. GCMS for essential oil showed 26 compounds including ±pinene. Leukocytes and erythrocytes of N. sativa and B. aristata were analyzed, and significant improvements were recorded (P < 0.05) and graphically presented. Mean survival time was calculated by the Kaplan Meier method (119 days). Immunoassay analyses were conducted, namely, TNF-α and VEGF, and interpreted and marked.

Intraocular pressure-lowering activity of topical application of Aegle marmelos fruit extract in experimental animal models.

AIM: The present study was designed to evaluate the intraocular pressure (IOP)-lowering activity of topical application of the aqueous extract of Aegle marmelos fruit in experimental animal models. MATERIALS AND METHODS: New Zealand white rabbits with normal and experimentally elevated IOP using water loading and steroid-induced models were included in this study. The IOP-lowering effect of A. marmelos fruit extract in rabbits with experimentally elevated IOP was also compared with that of timolol 0.25%. RESULTS: In rabbits with normal IOP, the A. marmelos fruit extract at a concentration of 1% showed the maximum IOP-lowering effect with 22.81% reduction from baseline IOP. The maximum IOP reduction achieved in water loading and steroid-induced models with the same concentration of A. marmelos was 27.57 and 28.41% from baseline, respectively. The efficacy was comparable to that of timolol after 45 min of water loading in the water loading model, and during the first 2 h of treatment in the steroid-induced model. CONCLUSION: A. marmelos fruit extract showed significant IOP-lowering activity in experimental animal models.

Advances in pharmacological strategies for the prevention of cataract development.

Cataractous-opacification of the lens is one of the leading causes of blindness in India. The situation can be managed by surgical removal of the cataractous lens. Various pharmacological strategies have been proposed for the prevention and treatment of cataract. Information on possible benefits of putative anticataract agents comes from a variety of approaches, ranging from laboratory experiments, both in vitro and in vivo , to epidemiological studies in patients. This review deals with the various mechanisms, and possible pharmacological interventions for the prevention of cataract. The article also reviews research on potential anticataractous agents, including aldose reductase inhibitors, glutathione boosters, antiglycating agents, vitamins and various drugs from indigenous sources.

Anti-implantation activity of H2 receptor blockers and meloxicam, a COX-inhibitor, in albino Wistar rats.

OBJECTIVE: To evaluate the anti-implantation activity of H(2) receptor blockers ranitidine and famotidine, and Cox-inhibitor meloxicam, alone and in combination, considering the role of histamine and prostaglandins in implantation. METHOD: The drugs were administered orally to female albino Wistar rats at different dose levels for 1 to 7 days, immediately after confirming copulation by observing sperm in the vaginal smear. A laparotomy was done on the 10th day of pregnancy, the implants and corpora lutea were counted, and the pre- and post implantation losses determined. The mast cell stabilising activity was studied using both in vitro and in vivo methods. RESULTS: Ranitidine 70 mg/kg (75.41%; p < 0.01), and famotidine 80 mg/kg (74.30%; p < 0.001) showed significant anti-fertility activity. No increase in activity was seen at higher doses. Meloxicam showed significant activity at doses of 3, 4, and 5 mg/kg. The combination of meloxicam (4 mg/kg) with ranitidine (70 mg) and famotidine (80 mg/kg) showed 100% anti-fertility activity. CONCLUSION: Our results indirectly confirm the combined involvement of histamine and prostaglandins in the implantation process. The mast cell stabilising property of H(2) blockers appears to be a possible mechanism for their anti-implantation activity.

Aqueous extract of Ficus religiosa linn. reduces oxidative stress in experimentally induced type 2 diabetic rats.

One of the major etiologies in pathogenesis of type 2 diabetes especially complications is oxidative stress. Aqueous extract of Ficus religiosa at a dose of 100 and 200 mg/kg orally decreased the fasting blood glucose in streptozotocin induced type 2 diabetic rats. The drug had enzyme induction effect with respect to catalase (CAT) and glutathione peroxidase (GSH-Px) activity, however decreased the exaggerated activity of superoxide dismutase (SOD) in type 2 diabetic rats. F. religiosa modulated the enzymes of antioxidant defence system to combat oxidative stress. As a result, glutathione (GSH-reduced form) was restored and inhibited the formation of malondialdehyde. Drug at higher dose (200 mg/kg) had more pronounced effect. F. religiosa, a rasayana group of plant drug having anti-diabetic activity along with antioxidant potential was beneficial in treatment of type 2 diabetes.

A novel polyherbal microbicide with inhibitory effect on bacterial, fungal and viral genital pathogens.

A polyherbal cream (Basant) has been formulated using diferuloylmethane (curcumin), purified extracts of Emblica officinalis (Amla), purified saponins from Sapindus mukorossi, Aloe vera and rose water along with pharmacopoeially approved excipients and preservatives. Basant inhibits the growth of WHO strains and clinical isolates of Neisseria gonorrhoeae, including those resistant to penicillin, tetracycline, nalidixic acid and ciprofloxacin. It has pronounced inhibitory action against Candida glabrata, Candida albicans and Candida tropicalis isolated from women with vulvovaginal candidiasis, including three isolates resistant to azole drugs and amphotericin B. Basant displayed a high virucidal action against human immunodeficiency virus HIV-1NL4.3 in CEM-GFP reporter T and P4 (Hela-CD4-LTR-betaGal) cell lines with a 50% effective concentration (EC50) of 1:20000 dilution and nearly complete (98-99%) inhibition at 1:1000 dilution. It also prevented the entry of HIV-1(IIIB) virus into P4-CCR5 cells (EC50 approximately 1:2492). Two ingredients, Aloe and Amla, inhibited the transduction of human papillomavirus type 16 (HPV-16) pseudovirus in HeLa cells at concentrations far below those that are cytotoxic and those used in the formulation. Basant was found to be totally safe according to pre-clinical toxicology carried out on rabbit vagina after application for 7 consecutive days or twice daily for 3 weeks. Basant has the potential of regressing vulvovaginal candidiasis and preventing N. gonorrhoeae, HIV and HPV infections.

Lowering of intraocular pressure by topical application of Daucus carota seed extract in rabbits.

In normotensive rabbits topical application of Daucus carota seed extract at the concentration of 0.3, 0.6 and 1.2% resulted in mean IOP reduction of 19.33. 23.20 and 25.61% respectively from baseline. As no significant difference was observed between the change in IOP in 0.6 and 1.2% extract treated groups, 0.6% concentration was chosen for further evaluation in rabbits with experimentally elevated IOP. In water loaded rabbits, maximum mean IOP reduction with 0.6% extract was 29.39%, which was comparable to pilocarpine. In steroid pretreated rabbits, maximum mean IOP reduction was 30.27% from baseline, which was significantly higher than pilocarpine. The extract showed a comparatively slower onset of action however, the duration of action was comparable to pilocarpine in all the experimental models.

Comparative efficacy of pilocarpine, timolol and latanoprost in experimental models of glaucoma.

Intraocular pressure (IOP)-lowering effects of investigational antiglaucoma drugs often need comparison with existing drugs, but detailed data showing comparative efficacy of antiglaucoma drugs with different mechanism of action has not been reported so far. This study was designed to establish baseline information of the IOP-lowering effect of three currently used antiglaucoma drugs in three experimental models in rabbits, so that they act as a benchmark for the efficacy evaluation of the future experimental antiglaucoma drugs. The IOP-lowering effect of single-drop application of pilocarpine, timolol and latanoprost was studied in normotensive, water loading and steroid-induced models of glaucoma in rabbits. The noncontact tonometer was used for the first time to estimate IOP in rabbits. The peak IOP-lowering effect of pilocarpine, timolol and latanoprost in normotensive rabbit eye was 18.23%, 20% and 22.56%, respectively. In water-loading model, the maximum protection against the rise in IOP was shown by latanoprost (40.27%), followed by timolol (31.39%) and pilocarpine (28.91%). In steroid-pretreated rabbit eyes, peak IOP-lowering effects of pilocarpine, timolol and latanoprost were 25.65%, 34.21% and 35.06%, respectively. Therefore, the latanoprost was found to be most effective in all three models followed by timolol and pilocarpine. The results of this study can be used for future preclinical investigations for the assessment of IOP-lowering activity of potential antiglaucoma drugs.

Estimation of intraocular pressure in rabbits using noncontact tonometer: a comparative evaluation with Schiotz tonometer.

Estimation of intraocular pressure in rabbits is often done by indentation tonometry using Schiotz tonometer. The use of Schiotz tonometer for estimation of intraocular pressure requires the use of topical anesthesia, poses risk of infection and injury to cornea and puts the animal under stress. Therefore, in this study noncontact tonometer was used to estimate intraocular pressure in rabbits and the technique is described for the first time. Twenty-five rabbits were subjected to intraocular pressure estimation using Schiotz as well as noncontact tonometer by two independent observers. All intraobserver and interobserver differences were within two standard deviation from the mean, indicating good repeatability and reproducibility of the two methods. A good linear correlation was observed between measurements done by Schiotz and noncontact tonometers with the correlation coefficient (r(2)) of 0.9122 and regression equation y = 1.632x + 4.8684. No significant difference was observed in percent change in intraocular pressure in timolol-treated, water-loaded rabbits using noncontact tonometer and Schiotz tonometer. Noncontact tonometer provides an easy, quick and safe method for intraocular pressure estimation, which is comparable to the conventional method of Schiotz tonometry. Therefore, despite the cost, noncontact tonometry seems to have a greater scope in experimental studies.

In vitro percutaneous absorption of verapamil.

The in vitro percutaneous absorption of verapamil hydrochloride (VHCl) was investigated in order to assess its feasibility for transdermal development. The experiments were carried across mice and guinea pig skins using Keshary-Chien diffusion cell. The values of diffusion rate (J) and permeability coefficient (Kp) across guinea pig skin were lowered as compared to mouse skin. Increased drug concentration in donor compartment increased value of J but decreased value of Kp. Under similar conditions, values of J and Kp were lowered for dorsal skin as compared to abdominal skin, both for mice and guinea pig. The results indicate that verapamil can be administered transdermally.

Hepatoprotective studies on Phyllanthus emblica Linn. and quercetin.

Phyllanthus emblica is a constituent of many hepatoprotective formulations available in market. 50% alcoholic extract of P. emblica and quercetin isolated from it were studied for hepatoprotective effect against country made liquor (CML) and paracetamol challenge in albino rats and mice respectively. The extract at the dose of 100 mg/100 g [corrected], po and quercetin at the dose of 15 mg/100 g, po, produced significant hepatoprotection.

In vitro transdermal delivery of atenolol using mouse and guinea pig.

In vitro percutaneous absorption of atenolol was done in order to assess its feasibility for transdermal development across mouse and guinea pig skins using Keshary-Chien type of diffusion cell. Values of diffusion rate (J) and permeability coefficient (Kp) across guinea pig skin were lowered as compared to those in mouse skin. When the concentration of drug in donor compartment was increased a decrease in Kp and increase in J value were observed with both the skins. Under the same conditions, values of J and Kp were lowered for dorsal skin compared to abdominal skin both for mouse and guinea pig. The results suggest that atenolol can be pursued further for transdermal system development.

Brucine, an indole alkaloid from Strychnos nux-vomica attenuates VEGF-induced angiogenesis via inhibiting VEGFR2 signaling pathway in vitro and in vivo.

In this study, we investigated the mechanism of brucine in tumor angiogenesis. We found that brucine inhibits VEGF-induced cell proliferation, chemotactic motility, and the formation of capillary-like structures in HUVECs in a dose-dependent manner. Brucine suppresses VEGF- induced p-VEGFR2 kinase activity and inhibits neovascularization in vivo. Brucine inhibits the downstream protein kinases of VEGFR2, including Src, FAK, ERK, AKT and mTOR. And further downregulates levels of VEGF, NO, IL-6, IL-8, TNF-α and IFN-γ in HUVECs. Taken together, our study suggests that brucine potently suppresses angiogenesis by targeting VEGFR2 activation and may be a viable drug candidate in anti-angiogenesis and anti-cancer therapies.

Ursolic acid inhibits tumor angiogenesis and induces apoptosis through mitochondrial-dependent pathway in Ehrlich ascites carcinoma tumor.

Ursolic acid (UA) is a pentacyclic triterpene naturally occurring in many plant foods. In the present study, we investigated anti-cancer activity of UA in vivo in Ehrlich ascites carcinoma (EAC) tumor. 15 × 10(6) EAC cells were implanted intraperitoneally (i.p., ascitic tumor) and subcutaneous (s.c., solid tumor) in Swiss albino mice. Mice with established tumors received UA i.p. at 25, 50 and 100mg/kg bw for 14 d in ascitic and 100mg/kg bw in solid tumor for 30 d. On day 15, blood samples were collected for hematological assessment of hemoglobin (Hb%), RBCs, WBCs and PCV. Tumor volume, cell viability, angiogenic, anti-angiogenic, anti-inflammatory factors and antioxidant parameters were determined. Immunohistochemistry analysis for VEGF, iNOS, CD31, caspase-3 and Bax were also performed. UA significantly inhibited tumor growth, cell viability, in both ascites and solid tumor model in vivo (p<0.001). The anti-angiogenic effects were accompanied with decreased VEGF, iNOS, TNF-α and increased IL-12 levels. UA at 100mg/kg bw dose significantly increased SOD and CAT activity (p<0.01). GSH and TBARS were increased as compared to control group (p<0.001). Furthermore, UA increased total RBCs, WBCs as well as Hb% significantly (p<0.05) compared to cyclophosphamide (CP). Histopathological examination of tumor cells in the treated group demonstrated signs of apoptosis with chromatin condensation and cell shrinkage. Decreased peritoneal angiogenesis showed the anti-angiogenic potential. UA downregulated VEGF & iNOS expression whereas bax and caspase-3 expressions were upregulated suggesting drug induced tumor cell apoptosis through activating the pro-apoptotic bcl-2 family and caspase-3 and downregulation of VEGF. The present study sheds light on the potent antitumor property of the UA and can be extended further to develop therapeutic protocols for treatment of cancer.

Punarnavine, an alkaloid from Boerhaavia diffusa exhibits anti-angiogenic activity via downregulation of VEGF in vitro and in vivo.

Punarnavine, a quinolizidine alkaloid isolated from Boerhaavia diffusa is known to possess analgesic, anti-inflammatory, hepato-protective, immunomodulatory and anti-proliferative properties. However, its roles in tumor angiogenesis and the involved molecular mechanism are still unknown. Therefore, we examined its anti-angiogenic effects and mechanisms in vitro and in vivo. We examined the effect of punarnavine on VEGF-A expression by RT-PCR, Western blotting and ELISA. In vivo antiangiogenic activity was determined using sponge implant angiogenesis assay and antitumor activity was evaluated against Ehrlich ascites carcinoma tumor. Punarnavine significantly inhibited endothelial cell migration and invasion and capillary structure formation of HUVECs. Punarnavine significantly at 50 µM inhibited MMP-2 and MMP-9 expression in HUVECs in vitro. Punarnavine inhibited neovascularization in sponge implant assay. Punarnavine (15 mg/kg bw/d) treatment showed dose-dependent decrease in the ascitic fluid volume by 60.94% and tumor volume by 86.40% in Ehrlich ascites model. Reduction in peritoneal angiogenesis with punarnavine treatment suggests the anti-angiogenic activity of punarnavine. The present study sheds light on the potent anti-angiogenic of the punarnavine and can be extended further to develop therapeutic protocols for treatment of cancer.

Anticarcinogenic effect of brucine in diethylnitrosamine initiated and phenobarbital-promoted hepatocarcinogenesis in rats.

We evaluated the effects of brucine on N-nitrosodiethylamine (DENA)-induced hepatocarcinogenesis in rats. Initiation of hepatocarcinogenesis was done by intraperitoneal injection of diethylnitrosamine (DENA) followed by promotion with phenobarbital. The rats were exposed to dietary brucine for 4 weeks prior to initiation, and the treatment was continued for 22 consecutive weeks. Brucine decreased the incidence, total number, multiplicity, size and volume of preneoplastic hepatic nodules in a dose-dependent manner. Administration of DENA induced hepatocellular carcinoma (HCC), as evidenced by changes in histopathological architecture, increased activity of cytochrome P450, decreased activity of glutathione Stransferase (GST) as well as decreased antioxidant status, enhanced lipid peroxidation, increased liver marker enzymes. Western blot analysis showed decreased expression of cyclin D1 and Bcl-2 with activation of caspase-3 and increased expression of Bax. Immunohistochemical demonstrated the decreased expression of the PCNA and VEGF. These results indicate that brucine prevents lipid peroxidation and hepatic cell damage and also protects the antioxidant system in DENA-induced hepatocarcinogenesis.