Chemical analysis of calabash chalk and its effect on locomotor activities and behavior in Swiss albino mice.

Continuous exposure of young women to calabash chalk, especially at child-bearing age, necessitated this study to analyze the chemical constituents of calabash chalk and to evaluate its effect on locomotor activities and behavior in Swiss albino mice. Dried cubes of calabash chalk were purchased and analyzed by atomic and flame atomic absorption spectrophotometer. Twenty-four Swiss-albino mice were taken and divided into four groups: control (1 ml distilled water) and three treated groups with the doses of 200 mg/kg, 400 mg/kg, and 600 mg/kg of calabash chalk suspension, respectively, by oral gavage. Hole Cross, Hole Board, and Open Field tests were performed to evaluate the locomotor activities, behavior, and anxiety along with measurement of body weight. The data were analyzed by SPSS-software. Chemical analysis of calabash chalk showed the presence of trace elements along with some heavy metals such as lead (19.26 ppm), Chromium (34.73 ppm), and arsenic (4.57 ppm). After 21 days of oral administration of calabash chalk, the study showed a significant decrease in body weight in the treated groups of mice (p < 0.01). Decreased locomotor activities were also observed in all three experiments. Significantly decreased locomotion and behaviors were also observed in the hole crossing, line crossing, head dipping, grooming, rearing, stretch attend, central square entry, central square duration, defecation, and urination, in a dose-dependent manner (p < 0.01). These effects also prove the anxiogenic behavior of calabash chalk in albino mice. Heavy metals are believed to be harmful to the brain and responsible for cognitive dysfunction and elevated anxiety. In this study, decreased body weight in mice may occur due to disorders in hunger and thirst centers of the brain by heavy metals. Therefore, heavy metals may be responsible for muscle weakness, decreased locomotor activities, and axiogenic effects of mice.

Therapeutic Potentials of Colocasia affinis Leaf Extract for the Alleviation of Streptozotocin-Induced Diabetes and Diabetic Complications: In vivo and in silico-Based Studies.

INTRODUCTION: Hypoglycemia in diabetes mellitus (DM) correlates with hepatic impairment, nephropathy, lipid abnormalities, and oxidative stress and subsequently complicates the disease pathogenesis. Medicinal plants have been used for the management of diabetes since ancient times. In this study, we explored the potentials of Colocasia affinis (CA), a plant known to possess anti-allergic and anti-inflammatory activities, as a remedy for diabetes and related complications. METHODS: We induced diabetes in rats using a single intraperitoneal dose (65 mg/kg) of streptozotocin (STZ). We next treated the rats with an ethanolic extract of leaves of CA to reveal its antidiabetic and organ-protective potentials. Biomarkers of diabetes, inflammation, and oxidative stress were measured using biochemical and histopathological analysis. We also performed molecular docking for three major phytochemicals (kaempferol, myricetin, and rosmarinic acid) of CA. RESULTS: Oral administration of the CA leaves extract at 250 mg/kg and 500 mg/kg doses decreased blood glucose level significantly (p<0.05) in STZ-induced diabetic rats. The extract also considerably attenuated plasma HbA1c levels and normalized blood lipids, glycogen, alanine aminotransferase (ALT), and aspartate aminotransferase (AST). Additionally, treatment with the extract improved kidney complications by decreasing serum creatinine and blood urea nitrogen (BUN) levels. Furthermore, CA leaves extract normalized nitric oxide (NO) and advance oxidative protein products (AOPP) in diabetic rats. The extract also showed significant improvement of the antioxidant enzymes glutathione dismutase (GSH) and superoxide dismutase (SOD) at a dose of 500 mg/kg. Besides, histological investigation demonstrated attenuation of inflammation of the vital organs, including the liver and the kidney. In silico studies revealed that three major phytochemicals (kaempferol, myricetin, and rosmarinic acid) of the ethanolic extract of leaves of CA can inhibit several molecular targets of diabetes and inflammation. CONCLUSION: Collectively, our results demonstrated the therapeutic potentials of CA for the mitigation of diabetes and diabetic complications.

Subacute oral toxicity of ayurvedic anti-diabetic preparation Jambadyarista in Sprague-Dawley rats.

BACKGROUND: Jambadyarista is an Ayurvedic polyherbal formulation widely prescribed by Ayurvedic practitioners for the management of diabetes and its associated complications. About 39 companies have marketed this formulation in Bangladesh with consent from the Directorate General of Drug Administration (DGDA). AIM: This study investigated the sub-acute oral toxicity of Jambadyarista in the Sprague-Dawley rat model. METHODS: The sub-acute toxicity studies were executed in Sprague-Dawley rats. Jambadyarista formulation was given for 28-days through oral gavage at 10 mL/kg and 20 mL/kg dose to two different groups comprising 6 rats of both sex/groups. Across the experimental period mortality, adverse reactions were closely monitored. After 28-day feeding hematological, biochemical, and relative organ weights were quantified. RESULTS: No mortality and/or signs of morbidity were observed for 28-day of repeated-dose sub-acute toxicity. Any pernicious change in body weight, biochemical, and hematological parameters along with relative organ weight were not observed for Jambadyarista. Correlation study among parameters of the renal profile, liver profile, lipid profile also metabolic hormones (T3 and TSH), and enzymes showed the non-toxic rather beneficial role (hypolipidemic) of Jambadyarista in Sprague-Dawley rats. CONCLUSION: Jambadyarista preparation did not cause any potential toxic effect in repeated dose subacute toxicity study over Sprague-Dawley rats orally. Therefore, low dose administration of Jambadyarista could have a beneficial effect on diabetes and can be considered safe before the chronic study.

Antioxidant and hepatoprotective activity of Piper retrofractum against Paracetamol-induced hepatotoxicity in Sprague-Dawley rat.

The ethanol extracts of Piper retrofractum were investigated for antioxidant and hepatoprotective activity. Hepatoprotective activity against paracetamol-induced acute hepatotoxicity was estimated in Sprague-Dawley rat. In DPPH free radical assay the root and stem extracts showed IC50 values at 133 and 91 µg/mL, respectively, while ascorbic acid at 14 µg/mL. Extracts also exhibited hydroxyl radical scavenging activity and reducing power. HPLC-DAD analysis indicated the presence of some polyphenolic compounds. Treatment of extracts significantly reduced the elevated serum levels of GPT (P < 0.01), GOT (P < 0.01) and bilirubin (P < 0.001). Both extracts restored the reduced level of total proteins and albumin. A significant increase in HDL-c but decrease in LDL-c level was observed compared to induced control. In histopathological study of liver sections, both extracts showed minimal to mild multifocal and diffuse granular degeneration and mild to moderate lobular disarray compared to control group. Results suggest that both extracts can prevent paracetamol induced hepatotoxicity.

Antihistamines suppress upregulation of histidine decarboxylase gene expression with potencies different from their binding affinities for histamine H1 receptor in toluene 2,4-diisocyanate-sensitized rats.

Antihistamines inhibit histamine signaling by blocking histamine H1 receptor (H1R) or suppressing H1R signaling as inverse agonists. The H1R gene is upregulated in patients with pollinosis, and its expression level is correlated with the severity of nasal symptoms. Here, we show that antihistamine suppressed upregulation of histidine decarboxylase (HDC) mRNA expression in patients with pollinosis, and its expression level was correlated with that of H1R mRNA. Certain antihistamines, including mepyramine and diphenhydramine, suppress toluene-2,4-diisocyanate (TDI)-induced upregulation of HDC gene expression and increase HDC activity in TDI-sensitized rats. However, d-chlorpheniramine did not demonstrate any effect. The potencies of antihistamine suppressive effects on HDC mRNA elevation were different from their H1R receptor binding affinities. In TDI-sensitized rats, the potencies of antihistamine inhibitory effects on sneezing in the early phase were related to H1R binding. In contrast, the potencies of their inhibitory effects on sneezing in the late phase were correlated with those of suppressive effects on HDC mRNA elevation. Data suggest that in addition to the antihistaminic and inverse agonistic activities, certain antihistamines possess additional properties unrelated to receptor binding and alleviate nasal symptoms in the late phase by inhibiting synthesis and release of histamine by suppressing HDC gene transcription.

Preliminary study of the immunostimulating activity of an ayurvedic preparation, Kanakasava, on the splenic cells of BALB/c mice in vitro.

CONTEXT: Immunostimulant plays an important role to prevent infections when defensive capacity of body is impaired, commonly occur with aging, cancer, diabetes, and sepsis. Kanakasava (KNK) is a polyherbal ayurvedic preparation used since ancient times for the treatment of respiratory diseases and to improve immunity. OBJECTIVE: The present study evaluated the immunostimulating potential of KNK. MATERIALS AND METHODS: The immunostimulating activity of KNK was evaluated by measuring immunoglobulin M (IgM) production and splenocyte proliferation in vitro. BALB/c mice splenocytes were treated with 0, 0.25, 0.5, 0.75, 1, 1.5, 2, 3, and 4% (v/v) of KNK, and the cells were subcultured at 37°C, humidified atmosphere containing 5% CO(2) for 120 h. The production of IgM in cultured supernatants were determined by an enzyme-linked immunosorbent assay (ELISA) and the proliferations of cells were measured by the 3-(4,5-dimethylthiazol-2-y)-2,5-diphenylterazolium bromide (MTT) method. RESULTS AND DISCUSSION: KNK at the doses of 0.25, 0.5, 0.75, 1, and 1.5% (v/v) significantly augmented polyclonal IgM production (1.211, 1.260, 1.274, 1.180, and 1.028 µg/mL, respectively) compared to control (0.246 µg/mL). Similarly, the same doses stimulated the proliferation of splenocytes as well (Abs. 0.270, 0.281, 0.368, 0.328, and 0.301, respectively, measured at 570 nm) compared to untreated cells (Abs. 0.137). The activity of KNK was not retarded by the treatment of cells with polymixin B. Thus, our results demonstrate that KNK possesses immunostimulating potential that acts through the induction of lymphocytes for proliferation and IgM production. CONCLUSION: KNK may be useful for strengthening immune responses in case of insufficient or impaired immunity.

Albizia lebbeck suppresses histamine signaling by the inhibition of histamine H1 receptor and histidine decarboxylase gene transcriptions.

Histamine plays major roles in allergic diseases and its action is mediated mainly by histamine H(1) receptor (H1R). We have demonstrated that histamine signaling-related H1R and histidine decarboxylase (HDC) genes are allergic diseases sensitive genes and their expression level affects severity of the allergic symptoms. Therefore, compounds that suppress histamine signaling should be promising candidates as anti-allergic drugs. Here, we investigated the effect of the extract from the bark of Albizia lebbeck (AL), one of the ingredients of Ayruvedic medicines, on H1R and HDC gene expression using toluene-2,4-diisocyanate (TDI) sensitized allergy model rats and HeLa cells expressing endogenous H1R. Administration of the AL extract significantly decreased the numbers of sneezing and nasal rubbing. Pretreatment with the AL extract suppressed TDI-induced H1R and HDC mRNA elevations as well as [(3)H]mepyramine binding, HDC activity, and histamine content in the nasal mucosa. AL extract also suppressed TDI-induced up-regulation of IL-4, IL-5, and IL-13 mRNA. In HeLa cells, AL extract suppressed phorbol-12-myristate-13-acetate- or histamine-induced up-regulation of H1R mRNA. Our data suggest that AL alleviated nasal symptoms by inhibiting histamine signaling in TDI-sensitized rats through suppression of H1R and HDC gene transcriptions. Suppression of Th2-cytokine signaling by AL also suggests that it could affect the histamine-cytokine network.

Suplatast tosilate inhibits histamine signaling by direct and indirect down-regulation of histamine H1 receptor gene expression through suppression of histidine decarboxylase and IL-4 gene transcriptions.

Allergic rhinitis (AR) is an inflammatory disorder typified by symptoms such as sneezing, congestion, and rhinorrhea. Histamine plays important roles in eliciting AR symptoms. Up-regulation of the histamine H(1) receptor (H1R) and histidine decarboxylase (HDC) mRNAs was observed in AR patients. Th2 cytokines are also involved in the pathogenesis of AR. We examined the effect of suplatast tosilate on nasal symptoms, and H1R, HDC, and IL-4 gene expression using toluene-2,4-diisocyanate (TDI)-sensitized rats and HeLa cells expressing endogenous H1R. Provocation with TDI increased nasal symptoms, HDC activity, the histamine content of nasal lavage fluid, and the expression of H1R, HDC, and IL-4 mRNAs in TDI-sensitized rats. Pretreatment with suplatast for 2 wk significantly suppressed TDI-induced nasal symptoms and elevation of H1R, HDC, and IL-4 mRNAs. Suplatast also suppressed HDC activity in the nasal mucosa and the histamine content of the nasal lavage fluid. Bilateral injection of IL-4 into the nasal cavity of normal rats up-regulated H1R mRNA, while intranasal application of histamine up-regulated IL-4 mRNA. Suplatast suppressed IL-4-induced up-regulation of H1R mRNA in HeLa cells. However, it did not inhibit histamine-induced H1R mRNA elevation. These results suggest that suplatast alleviates nasal symptoms by inhibiting histamine signaling in TDI-sensitized rats through the suppression of histamine- and IL-4-induced H1R gene expression by the inhibitions of HDC and IL-4 gene transcriptions, respectively.

Clofarabine induces hypomethylation of DNA and expression of Cancer-Testis antigens.

In this study, treatment of lymphoid tumor cells with low dose clofarabine upregulated the expression of Sp17 and SPAN-Xb. This was associated with an increase in hypomethylated CpG dinucleotides and a decrease in global DNA methylation, as demonstrated by decreases in the percent of methylated Alu repeats. The most optimal concentration of clofarabine to induce DNA hypomethylation and CT antigen expression was between 1x10(-9) and 1x10(-8)M. Above this, clofarabine resulted in tumor cell growth inhibition and apoptosis. Our results provide the first evidence for the CT antigen-inducing and DNA hypomethylating property of low concentration clofarabine.

SEMG-1 expression in early stage chronic lymphocytic leukemia.

BACKGROUND AIMS: Chronic lymphocytic leukemia (CLL) is an indolent disease. It is currently recommended that patients with CLL stages 0 and I follow a watchful waiting strategy. These patients are, therefore, a suitable group for testing immunotherapeutic approaches to avoid problems of immunosuppression as a result of disease progression and chemotherapy. In this study, we investigated the expression of SEMG-1 in early CLL to determine the suitability of SEMG-1 as a target for further development of tumor vaccines for early CLL. METHODS: A combination of reverse transcriptase (RT)-polymerase chain reaction (PCR) and immunocytochemistry was used to evaluate the expression of SEMG-1 in early CLL. The results were correlated with Zap 70 expression. Recombinant SEMG-1 protein was used in an enzyme-linked immunosorbent assay (ELISA) to determine the presence of SEMG-1 antibodies (Ab) in serum from these patients. RESULTS: The SEMG-1 gene was expressed in 19/41 (46%) patients with early CLL. Gene expression was associated with protein synthesis in CLL cells. Protein expression, however, was heterogeneous within individual patients. Only transcripts encoding the SEMG-1(50) variant and not SEMG-1(43) were detected. SEMG-1(50) was expressed irrespective of the Zap 70 status. High-titer SEMG-1 IgG but not IgM Ab were detected in some of these patients, suggesting that SEMG-1-reactive immune responses are intact within the immune repertoire of early CLL patients. CONCLUSIONS: SEMG-1 is expressed in nearly half of patients with early CLL and may be a target for further investigations into its use for immunotherapy of early CLL.

Identification of protamine 1 as a novel cancer-testis antigen in early chronic lymphocytic leukaemia.

Early chronic lymphocytic leukaemia (CLL) is an ideal disease for immunotherapy. We previously showed that SEMG 1 is a cancer-testis (CT) antigen in CLL. In this study, SEMG 1 was applied as the bait in a yeast two-hybrid system of a testicular cDNA library. Seven clones were isolated and Protamine (Prm) 1 was identified as a novel CT antigen in early CLL. PRM1 transcripts were detected in 11/41 (26.8%) patients. Prm 1 protein was also expressed but heterogeneously within individual patients. Of the 11 patients expressing Prm 1, four expressed Zap 70 protein and seven did not. These results, therefore, indicate that Prm 1 could potentially be a suitable target for the design of tumour vaccine for patients with early CLL, including for those with poor risk CLL. High titres of Prm 1 IgG antibodies could be detected in 20 of these 41 CLL patients but not in any of the 20 healthy donors (P = 0.0001), suggesting the presence of Prm 1-reactive immune responses within the immune repertoire of patients with early CLL. Further work is warranted, especially in approaches to upregulate Prm 1 expression, and to determine the role of Prm 1 as an immunotherapeutic target for early CLL.