Fragility fracture care gap at a tertiary teaching hospital in Malaysia.

Fracture begets fracture, pharmacological treatment is needed to prevent secondary fractures. This study found that there was a fragility fracture care gap where both bone health investigations and treatment initiation rates were low. Strategies such as Fracture Liaison Service is needed to address the care gap. PURPOSE: This study aimed to investigate the clinical burden and secondary fracture prevention of fragility fractures at a tertiary teaching hospital in Malaysia. METHODS: Electronic medical records of all patients admitted with fragility fractures between 1 January 2017-31 December 2018 were reviewed. Patients < 50 years old, with non-fragility fractures, restricted access to medical records, transferred to another hospital or who passed away during admission were excluded. Descriptive statistics were used to summarise patients' characteristics, frequency of fragility fractures, and secondary fracture prevention details. Binomial logistic regression was performed to analyse predictive factors for post-fracture bone health assessments and treatment initiation. RESULTS: 1030 patients [female (767/1030, 74.5%)] presented with 1071 fractures [hip fractures (378/1071, 35.3%)]. 170/993 (17.1%) patients were initiated on anti-osteoporosis medications (AOMs) and 148/984 (15.0%) had bone mineral density (BMD) performed within 1-year post-fracture. Less than half (42.4%) of the patients remained on treatment at 1-year post-fracture. Older patients [65-74 years old: odds ratio (OR) = 2.18, 95%CI 1.05-4.52, p = 0.04; ≥ 75 years: OR = 3.06, 95%CI 1.54-6.07, p < 0.01], hip fractures (OR = 1.95, 95%CI 1.23-3.11, p < 0.01), Chinese ethnicity (OR = 1.90, 95%CI 1.07-3.35, p = 0.03),previously diagnosed with osteoporosis (OR = 2.65, 95%CI:1.32-5.31, p < 0.01) and a BMD test performed (OR = 12.48, 95%CI 8.04-19.37, p < 0.01) were found to have higher AOM initiation. Patients with past diagnosis of osteoporosis (OR = 4.45, 95%CI 2.25-8.81, p < 0.01) and initiated on AOM (OR = 11.34, 95%CI 7.57-16.97, p < 0.01) had a higher likelihood to undergo BMD testing. CONCLUSION: The AOM initiation and BMD testing rates were low. There is a need to address the fragility fracture care gap with strategies such as Fracture Liaison Service.

Hip fracture management in the emergency department and its impact on hospital outcomes: a retrospective cross-sectional analysis.

PURPOSE: The emergency department (ED) plays an important role in initiating early treatment for hip fractures and ensuring prompt transfer to orthopaedic wards. This study reported on the care delivered in a tertiary centre ED in Malaysia and the association between time spent in ED with hospital outcomes. METHODS: Patients aged ≥ 65 years with fragility hip fractures and seen by the geriatric team were recruited. Data were collected on patient characteristics, key time points for treatment and hospital outcomes. Median time in ED was used to dichotomise long and short waiting time. RESULTS: 447 patients were recruited. The mean (SD) age was 80.5 (7.0) years and 69.8% were women. 74.9% were prescribed analgesia within 30 min. Median (Q1,Q3) time to diagnostic imaging was 27.0 (24.0-43.0) minutes, clinician confirmation of fracture was 83.0 (49.0-129.0) minutes, and time in ED was 4.8 (3.5-6.9) h. A weekday, weekend, in-hour or out-of-hour admission did not demonstrate a difference in the time important care was delivered. Patients who spent ≥ 5 h in ED had more cardiac events (4.6 vs 10.1%, p = 0.023) and more spent ≥ 14 days in hospital (17.5 vs 29.0%, p = 0.004) compared to those < 5 h. No significant increase in inpatient complications (43.5 vs 34.6%, p = 0.054), length of stay (median, 8 vs 7 days, p = 0.119), care home discharge (5.3 vs 4.6%, p = 0.772), or in-hospital death (6.3 vs 4.2%, p = 0.313) were observed. CONCLUSION: Time to early hip fracture pain relief and diagnosis was adequate in this ED. Time ≥ 5 h in ED was associated with cardiac events and 2 weeks or more inpatient stay.

Mutagenicity and Acute Oral Toxicity Test for Herbal Poultry Feed Supplements.

Herbal products are being used and trusted globally for thousands of years for their health benefits and limited side effects. Globally, a general belief amongst the consumers is that herbal supplements are always safe because they are "natural." But later, research reveals that they may not be safe. This raises concern on their safety and implications for their use as feed supplement or medicine. Toxicity testing can reveal some of the risks that may be associated with use of herbs, therefore avoiding potential harmful effects. The present study was designed to investigate five poultry feed supplements (PFS), EGMAX® (to revitalize ovarian activity), FEED-X™ (feed efficiency enhancer), KOLIN PLUS™ (natural replacer of synthetic choline chloride), PHYTOCEE® (natural defence enhancer), and STODI® (to prevent and control loose droppings), for their possible mutagenicity and toxicity. Bacterial reverse mutation (BRMT) and acute oral toxicity tests were employed to assess the PFS for their possible mutagenicity and toxicity. Results indicated that the PFS were devoid of mutagenic effects in BRMT and showed higher safety profile in rodent acute oral toxicity test.

Alleviation of Heat Stress by a Polyherbal Formulation, Phytocee™: Impact on Zootechnical Parameters, Cloacal Temperature, and Stress Markers.

BACKGROUND: The range of thermoneutral zone of chickens is narrow, and they become easily susceptible to environmental stress, a common and major concern for poultry causing a production loss. OBJECTIVE: The present study was designed to comparatively evaluate anti-stress activity of Phytocee™ and Vitamin C in chickens reared under heat stress. MATERIALS AND METHODS: A total of 600-day-old chicks of Cobb 400 were randomly assigned to 4 groups with 6 replicates comprising 25 birds each (n = 150). G1 served as a normal control (NC) and supplemented with Vitamin C at 100 g/ton of feed. G2 served as a heat stress control (HSC), subjected to heat stress (34°C-36°C) without Vitamin C supplementation. G3 and G4 served as positive control and treatment group (TC), subjected to heat stress and supplemented with Vitamin C and Phytocee™ at 100 g/ton of feed, respectively. The impact on zootechnical parameters and cloacal temperature was assessed at regular intervals, and blood was collected at the end of the experiment for evaluation of stress parameters, namely heterophil lymphocyte ratio (H:L ratio) and serum corticosterone. RESULTS: Exposure of chickens to heat stress caused a significant decrease in body weight, worsening of feed conversion ratio, higher mortality, and poor production efficiency. Moreover, serum corticosterone level, H:L ratio, and cloacal temperature were significantly increased in HSC as compared to NC. However, supplementation of Phytocee™ in feed significantly ameliorated the negative impact of heat stress in broiler birds. CONCLUSION: The supplementation of Phytocee™ demonstrated an anti-stress effect in chickens through restoration of serum corticosterone level, H:L ratio, and thermoregulatory mechanism. SUMMARY: Combating heat stress remains a challenge for the broiler industry in the tropics and subtropics, which is even aggravated by the changing climatic conditionsThe present study was designed to evaluate the anti-stressor activity of Phytocee™, a polyherbal formulation containing Emblica officinalis, Ocimum sanctum, and Withania somnifera in broiler using heat stress model in comparison with Vitamin CPhytocee™ demonstrated an anti-stress effect in the current study by ameliorating the negative effects of heat stress on zootechnical parameters, serum corticosterone, heterophil lymphocyte ratio, and cloacal temperature of broilers through modulating the hypothalamic-pituitary-adrenal axis and thermoregulatory mechanismHence, Phytocee™ could be recommended in broilers and livestock animals for modulating and combating adverse effects of heat stress and thereby reducing the economic losses incurred by farmers. Abbreviations Used: HPA axis: Hypothalamic pituitary adrenal axis.

Antiarthritic Effect of Polar Extract of Curcuma longa on Monosodium Iodoacetate Induced Osteoarthritis in Rats.

BACKGROUND: Curcuma longa Linn, "the golden spice" is a common spice used in Southern Asia and Middle East countries. It has a history of ethnopharmacological use for its various activities like anti-septic, anti-inflammatory, anti-oxidant, anti-microbial, anti-cancer and so on. OBJECTIVE: To investigate the effects of polar extract of C. longa (PCL) against monosodium iodoacetate (MIA) induced osteoarthritis in rat and to compare with curcuminoids, which are contemporarily believed to be the only active phytochemicals of C. longa for relieving pain in osteoarthritis. METHOD: Osteoarthritis in rats was induced by intra-articular injection of monosodium iodoacetate (MIA) in right knee. PCL or curcuminoids or tramadol was administered orally as single dose on the 5th day post MIA injection to rats. Weight bearing capacity and percentage inhibition of nociception of PCL treated groups were determined and compared with curcuminoids and tramadol (reference drug). In addition, gene expression levels of type II collagen and matrix metalloproteinases (MMP) in joint cartilage was measured by Reverse transcription polymerase chain reaction. RESULTS: PCL significantly decreased the difference in weight distribution between left and right limb in a dose dependent manner. Anti-arthritic activity of PCL is evident from significant up regulation of type II collagen gene (COL2A1) and down regulation of MMP-3 and MMP-7. CONCLUSION: Polar extract of C. longa showed beneficial effects on joints by exhibiting antiosteoarthritic effects via maintaining equilibrium between anabolic and catabolic factors of joint cartilage.

Evaluation of subacute toxicity of methanolic/aqueous preparation of aerial parts of O. sanctum in Wistar rats: Clinical, haematological, biochemical and histopathological studies.

ETHNOPHARMACOLOGICAL RELEVANCE: Ocimum sanctum, commonly known as Holy Basil or Tulsi has been used in Ayurveda as a demulcent, stimulant, expectorant; in the treatment of bronchitis, skin infections, malaria, diarrhoea, dysentery, arthritis, gastric and inflammatory disorders. We have previously shown that methanolic/aqueous extract of O. sanctum did not induce genotoxicity and other toxic effects in acute oral toxicity study. In the present report, we have performed sub-acute toxicity of methanolic/aqueous preparation of O. sanctum in Wistar rats to evaluate whether it induced any chronic toxic effects. MATERIALS AND METHODS: In subacute toxicity study, animals received O. sanctum extract (OSE) by oral gavage at the doses of 250, 500 and 1000 mg/kg/day (n=5/group/sex) for 28 days. At the end of the study, the animals were sacrificed and evaluated for the effect of OSE on clinical, haematological, biochemical and histopathological parameters. RESULTS: The rats treated with OSE did not show any change in body weight, food and water consumption, motor activity, sensory reactivity and foot splay measurements. There were no significant changes in haematological, pathological and biochemical parameters; and histopathology of tissues (liver, kidney, spleen, heart, and testis/ovary) among rats of either sex. OSE at a dose of 1000 mg/kg showed significant increase of Mean corpuscular hemoglobin (MCH) (19.8 ± 0.8; 18.7 ± 0.5) and mean corpuscular hemoglobin concentration (MCHC) (41.8 ± 1.1; 39.3 ± 0.7) in male and female rats in comparison to their respective controls (MCH: 17.7 ± 0.3; 17.4 ± 0.3; MCHC: 37.8 ± 0.5; 36.1 ± 0.2). Urine parameters (appearance, blood, nitrate, leucocyte, glucose, ketone, pH, protein and specific gravity) in both the male and female rats were comparable to their respective controls. In addition, no changes were observed in the vital organs of rats at macroscopic and microscopic levels. CONCLUSIONS: Our results showed that oral administration of OSE was not toxic to male and female Wistar rats upto the highest dose tested, thereby suggesting its clinical usefulness.

Anti-Inflammatory Activity of Polysaccharide Fraction of Curcuma longa Extract (NR-INF-02).

The aim of the study was to investigate the safety and anti-inflammatory effects of polysaccharide fraction (F1) of Curcuma longa extract (NR-INF-02) in classical rodent models of inflammation. F1 was evaluated for its acute oral toxicity and found to be safe upto 5000 mg/kg body weight in rats. The anti-inflammatory activity of F1 was evaluated in acute (carrageenan - induced paw edema; xylene - induced ear edema) and chronic (cotton pellet - induced granuloma) models of inflammation. The results of the study demonstrated that F1 significantly (p ≤ 0.05) inhibited carrageenan-induced paw edema at 1 h and 3 h at doses of 11.25, 22.5 and 45 mg/kg body weight in rats. Also, F1 at doses of 15.75, 31.5 and 63 mg/kg significantly inhibited the xylene induced ear edema in mice. In a chronic model, F1 at 11.25, 22.5 and 45 mg/kg doses produced significant reduction of wet and dry weights of cotton pellets in rats. Overall results indicated that F1 of NR-INF-02 significantly attenuated acute and chronic inflammation in rodent models. This study emphasizes on the importance of Curcuma longa polysaccharide's role in acute and chronic inflammation.

Pharmacokinetics of rifabutin during atazanavir/ritonavir co-administration in HIV-infected TB patients in India.

SETTING: Rifabutin (RBT) is reported to be as effective as and to have less inducing effect on cytochrome P450 enzymes than rifampicin against tuberculosis (TB). The optimal dose of RBT during ritonavir (RTV) co-administration remains a matter of debate. OBJECTIVE: To study the pharmacokinetics of 150 mg RBT thrice weekly during concomitant atazanavir/RTV administration in human immunodeficiency virus (HIV) infected TB patients. METHODS: This observational study was conducted in 16 adult HIV-infected TB patients being treated for TB with an RBT-containing regimen and an antiretroviral therapy regimen with RTV; the dose of RBT was 150 mg thrice weekly. Serial blood draws were performed at pre-dosing and at 1, 2, 4, 6, 8, 12 and 24 h after the drug was administered. Plasma RBT was estimated using high-performance liquid chromatography. RESULTS AND CONCLUSIONS: Peak RBT concentration was below the lower therapeutic limit (<0.3 µg/ml) in seven patients, while 10 patients had trough concentrations below the minimal inhibitory concentration against Mycobacterium tuberculosis (0.06 µg/ml), suggesting that the RBT dosage may be inadequate. Prospective studies in different settings are required to arrive at the proper therapeutic dose for RBT to be used during co-administration with RTV.

Evaluation of the mutagenic potential and acute oral toxicity of standardized extract of Ocimum sanctum (OciBest™).

Ocimum sanctum L. (Lamiaceae) is found throughout India and in many parts of world. O. sanctum is used for the treatment of various health indications. In this lieu, it is of prime importance to investigate the safety aspects of the plant. Hence, the present study was conducted to investigate the possible genotoxic potential and acute oral toxicity of the extract of O. sanctum (OciBest™). The standard battery of in vitro genotoxicity tests, namely bacterial reverse mutation, chromosome aberration and micronucleus (MN) tests were employed to assess the possible mutagenic activity. The results showed that OciBest™ (7.9-2500.0 µg/mL) did not increase the number of histidine revertant colonies in Salmonella typhimurium strains (TA98 and TAMix) with and without exogenous metabolic activation (S9). OciBest™ (10.0-100.0 µg/mL) did not show structural chromosomal aberrations or increase in MN induction, with and without S9, at the tested dose range in both 4-h and 18-h exposure cell cultures. Thus, OciBest™ is not genotoxic in bacterial reverse mutation, chromosomal aberration and MN tests. In an acute oral toxicity test, rats were treated with 5 g/kg of OciBest™ and observed for signs of toxicity for 14 days and the results did not show any treatment-related toxic effects to Wistar rats.

In vitro comparative evaluation of non-leaves and leaves extracts of Andrographis paniculata on modulation of inflammatory mediators.

This study was designed to evaluate and compare the inhibitory property of extracts of Andrographis paniculata leaves [methanolic (AP1), hydroalcoholic (AP2), successive water (AP3)] and non-leaves [methanolic (AP4), hydroalcoholic (AP5), successive water (AP6)] towards inflammatory mediators (NO, IL-1 beta, IL-6, TNF alpha, PGE2, TXB2 and LTB4). Stimulant induced J774A.1 murine macrophages and HL-60 promyelocytic leukemic cells were used to study the inhibitory potential of extracts of A. paniculata on inflammatory mediators. Results revealed that AP1 and AP4 exhibited inhibitory effect on all the inflammatory mediators excluding PGE2 and TNF-alpha. AP2 and AP5 exhibited inhibitory effect towards IL-1 beta, TXB2 and did not show inhibitory effect towards other mediators. However, AP3 and AP6 failed to show inhibitory activity against any of the inflammatory mediators at the tested concentrations. Further, we observed that the magnitude of inhibitory effect displayed by A. paniculata extracts depends on the andrographolide content, although, it does not appear to influence the inhibitory effect towards LTB4 production.

Hepatoprotective activity of Indian Phyllanthus.

CONTEXT: Phyllanthus (Euphorbiaceae) species are traditionally well-known for their medicinal properties including hepatoprotective activity. OBJECTIVE: The study assessed the hepatoprotective and antioxidant activities of 11 Phyllanthus species, P. amarus Schumach., P. urinaria L., P. debilis Klein ex Willd, P. tenellus Roxb., P. virgatus G. Forst., P. maderaspatensis L., P. reticulatus Poir., P. polyphyllus Willd., P. emblica L., P. indofischerii Bennet. and P. acidus (L.) Skeels. MATERIALS AND METHODS: The dried leaves and stems of each plant species were extracted in methanol and successively in water. The extracts were screened for hepatoprotective activity at a concentration of 50 µg/mL against tert-butyl hydroperoxide (t-BH) induced toxicity in HepG2 cells. Seven extracts from five species that showed hepatoprotective activity were assessed for their 50% effective concentration (EC50) values and their antioxidant activity using a DPPH assay. Phyllanthin and hypophyllanthin contents were also determined in these Phyllanthus species. RESULTS: The methanol extracts of P. polyphyllus, P. emblica and P. indofischeri showed high levels of hepatoprotective activity with EC50 values of 12, 19 and 28 µg/mL and IC50 of 3.77, 3.38 and 5.8 µg/mL for DPPH scavenging activity respectively against an IC50 of 3.69 µg/mL for ascorbic acid. None of these activities could be attributed to phyllanthin and hypophyllanthin. DISCUSSION AND CONCLUSION: The hepatoprotective and antioxidant activities of P. indofischeri are demonstrated for the first time in literature. The study also confirms the hepatoprotective and antioxidant activities of leaves of P. emblica and P. polyphyllus. The molecule(s) responsible for the activities is being investigated.

Evaluation of the genotoxic potential of standardized extract of Glycyrrhiza glabra (GutGard™).

Glycyrrhiza glabra Linn. (licorice) is widespread throughout the Mediterranean region and certain areas of Asia. Historically, the dried rhizome and root of the plant were used by the Chinese, Egyptian, Greek, Indian, and Roman civilizations as expectorant and carminative. In the modern medicinal system, licorice is used to treat liver ailments, dyspepsia, bronchitis, rheumatoid arthritis etc. Despite the extensive pharmacological applications, the genotoxic potential of G. glabra extract (GutGard™) has not been evaluated. Hence, this study was conducted to investigate the genotoxic potential of GutGard™ using battery of in vitro test systems: bacterial reverse mutation test (Ames II™), chromosome aberration (CA) and micronucleus (MN) tests. GutGard™ did not show significant increase in number of revertant colonies in Salmonella typhimurium strains (TA98 and TAMix) with/without S9 fraction. In CA and MN studies, GutGard™ did not show clastogenic effect at 4 and 18 h treatments with and without S9 fraction. Results indicated that GutGard™ is not mutagenic in a battery of genotoxicity tests.

Modulation of lipopolysaccharide-induced pro-inflammatory mediators by an extract of Glycyrrhiza glabra and its phytoconstituents.

OBJECTIVE: To evaluate the inhibitory property of de-glycyrrhizinated extract of Glycyrrhiza glabra root and its phytoconstituents (glabridin, isoliquiritigenin and glycyrrhizin) on LPS-induced production of pro-inflammatory mediators. MATERIALS AND METHODS: Inhibitory effect of G. glabra extract and its phytoconstituents were studied on lipopolysaccharide (LPS)-induced nitric oxide (NO), interleukin-1 beta (IL-1 beta) and interleukin-6 (IL-6) levels in J774A.1 murine macrophages. RESULTS: G. glabra and isoliquiritigenin significantly inhibited LPS stimulated NO, IL-1 beta and IL-6 production. Glabridin showed significant inhibition of NO and IL-1 beta release, but failed to attenuate IL-6 levels at the tested concentrations. In addition, glycyrrhizin did not exhibit inhibitory response towards any of the LPS-induced pro-inflammatory mediators at the tested concentrations. CONCLUSION: From the results we speculate that the inhibitory effect of G. glabra extract on LPS-induced pro-inflammatory mediators is influenced by glabridin and isoliquiritigenin and is not contributed by glycyrrhizin.

In vitro modulation of LPS/calcimycin induced inflammatory and allergic mediators by pure compounds of Andrographis paniculata (King of bitters) extract.

The aim of the current study is to probe the anti-inflammatory/anti-allergic potential of seven phytoconstituents (andrographolide, neoandrographolide, isoandrographolide, andrograpanin, 14-deoxy-11,12-didehydroandrographolide, 7-O-methylwogonin and skullcapflavone-I) isolated from Andrographis paniculata (King of bitters) on the production of key inflammatory/allergic mediators (NO, PGE(2), IL-1 beta, IL-6, LTB(4), TXB(2) and histamine). The results demonstrated that andrographolide, isoandrographolide, 7-O-methylwogonin and skullcapflavone-I significantly inhibited LPS stimulated NO and PGE(2) release in J774A.1 macrophages. Andrographolide, isoandrographolide and 7-O-methylwogonin showed considerable inhibition of IL-1 beta production in LPS elicited macrophages. LPS induced IL-6 production was significantly inhibited by andrographolide, isoandrographolide and skullcapflavone-I in a concentration dependent manner. The results revealed that andrographolide, isoandrographolide and skullcapflavone-I significantly decreased TXB(2) release in A23187 activated HL-60 promyelocytic cells. Furthermore, the anti-allergic properties of the phytoconstituents was investigated on A23187 induced LTB(4) production (HL-60 cells) and histamine release (RBL-2H3 basophilic cells). The results showed that only skullcapflavone-I and 7-O-methylwogonin showed marked inhibitory effect on LTB(4) production, however, only 7-O-methylwogonin exerted dose-dependent inhibition towards histamine release. Therefore, this study indicates that some of these phytoconstituents exhibit potent anti-inflammatory/anti-allergic effects by modulating different inflammatory/allergic mediators. Hence, these phytoconstituents might provide useful phytomedical treatment against variety of inflammatory and allergic disorders.

Dual inhibitory effect of Glycyrrhiza glabra (GutGard™) on COX and LOX products.

Glycyrrhiza glabra and its phytoconstituents have been known to possess widespread pharmacological properties as an anti-inflammatory, anti-viral, antitumour and hepatoprotective drug. In this study, we examined the inhibitory potential of extract of G. glabra (GutGard™) root and its phytoconstituents (glabridin, glycyrrhizin, and isoliquiritigenin) on both cyclooxygenase (COX) and lipoxygenase (LOX) products in order to understand the mechanism of its anti-inflammatory action. Inhibitory effect of GutGard™ and its phytoconstituents on lipopolysaccharide (LPS) induced prostaglandin E(2) (PGE(2)), calcimycin (A23187) induced thromboxane (TXB(2)), and leukotriene (LTB(4)) release was studied using murine macrophages (J774A.1) and human neutrophil (HL-60) cells. Results revealed that, G. glabra and glabridin significantly inhibited PGE(2), TXB(2) (COX) and LTB(4) (LOX), while, isoliquiritigenin exerted inhibitory effect only against COX products but failed to suppress LOX product. However, glycyrrhizin at the tested concentrations failed to exhibit inhibitory effect on both COX and LOX products. Here, we report for the first time that G. glabra (almost devoid of glycyrrhizin) exhibits anti-inflammatory property likely through the inhibition of PGE(2), TXB(2) and LTB(4) in mammalian cell assay system, which could be influenced in part by glabridin and isoliquiritigenin.

Optimization of cell-based assays to quantify the anti-inflammatory/allergic potential of test substances in 96-well format.

OBJECTIVE: There is an insistent need for robust, reliable, and optimized assays for screening novel drugs targeting the inflammatory/allergic markers. The present study describes about the optimization of eight cell-based assays utilizing mammalian cell lines in 96-well format for quantifying anti-inflammatory/allergic drug candidates. MATERIALS AND METHODS: We estimated the inhibitory response of reference compounds: 1400 W dihydrochloride on LPS-induced NO release, celecoxib on LPS-induced PGE(2) production and dexamethasone on LPS-induced pro-inflammatory cytokines IL-1 beta, IL-6, and TNF-alpha production by J774A.1 murine macrophages. Response of acetylsalicylic acid and celecoxib was studied on A23187-induced TXB(2) production; captopril on A23187-stimulated LTB(4) production by HL-60 cells. Effect of ketotifen fumarate was evaluated on A23187-elicited histamine release by RBL-2H3 cells. Each experiment was repeated twice to assess the reproducibility and suitability of the assays by determining appropriate statistical tools viz. %CV, S/B and Z' factor. RESULTS: 1400 W dihydrochloride was capable of inhibiting LPS-induced NO levels (IC(50) = 10.7 µM). Dexamethasone attenuated LPS-induced IL-1 beta (IC(50) = 70 nM), IL-6 (IC(50) = 58 nM) and TNF-alpha (IC(50) = 44 nM) release, whereas celecoxib, a specific COX-2 inhibitor showed marked reduction in LPS-induced PGE(2) (IC(50) = 23 nM) production. Captopril (IC(50) = 48 µM) and ketotifen fumarate (IC(50) = 36.4 µM) demonstrated potent inhibitory effect against A23187-stimulated LTB(4) and histamine levels, respectively. Both acetylsalicylic acid (IC(50) = 5.5 µM) and celecoxib (IC(50) = 7.9 nM) exhibited concentration-dependent decrease in TXB(2) production. Results for all the cell assays from two experiments showed a Z' factor varying from 0.30 to 0.99; the S/B ratio ranged from 2.39 to 24.92; %CV ranged between 1.52 and 20.14. CONCLUSION: The results proclaim that these cell-based assays can act as ideal tools for screening new anti-inflammatory/anti-allergic compounds.

Effect of an extract of Andrographis paniculata leaves on inflammatory and allergic mediators in vitro.

AIM OF STUDY: Andrographis paniculata has been known to possess widespread traditional application in the treatment of allergy and inflammatory diseases. In the current study, we sought to examine the effects of an extract of Andrographis paniculata leaves on inhibition of lipopolysaccharide (LPS) induced [nitric oxide (NO), prostaglandin E(2) (PGE(2)), interleukin-1beta (IL-1 beta), and interleukin-6 (IL-6)] and calcimycin (A23187) induced [leukotriene B(4) (LTB(4)), thromboxane B(2) (TXB(2)) and histamine] mediators in diverse cell based models. MATERIALS AND METHODS: Effect of an extract of Andrographis paniculata leaves (AP) was studied on inhibition of LPS induced NO, PGE(2), IL-1 beta and IL-6 in J774A.1 murine macrophages; A23187 induced LTB(4) and TXB(2) in HL-60 promyelocytic leukemic cells and histamine in RBL-2H3 rat basophilic leukemia cells. RESULTS AND CONCLUSION: AP illustrated significant alleviation of pro-inflammatory, inflammatory, and allergic mediators. However, no inhibition was observed against histamine release. This outcome has been summed up to deduce that AP is fairly potent in attenuating the inflammation by inhibiting pro-inflammatory (NO, IL-1 beta and IL-6), inflammatory (PGE(2) and TXB(2)) and allergic (LTB(4)) mediators.

In vitro efficacy and safety of poly-herbal formulations.

Indigenous plants are used as a traditional source of raw materials for the manufacture of medicines. Modernizing the ancient art of herbal medicine bequeathed from generations entails addressing two interrelated issues i.e. efficacy, and safety prior to their acceptance and use worldwide. The present study was designed to investigate three of our veterinary poly-herbal formulations - Phytocee an antistressor; Zigbir(R) a hepatoprotectant; and Zist(R) as an immunomodulator in the pertinent in vitro cell assay models in order to validate their therapeutic potential. Cellular antioxidant potential of Phytocee was demonstrated against AAPH induced oxidative stress using HepG2 cells. Zigbir(R) was confirmed as a hepatoprotectant against tert-butylhydroperoxide induced cytotoxicity in HepG2 cells. Immunomodulatory activity of Zist(R) was established by its ability to inhibit the proliferation of mitogen stimulated murine splenocytes in vitro. On treatment with Zist(R), a trend of decline in IL-6, and IL-12 levels was observed following stimulation with Con A, and LPS respectively in murine splenocytes. Further, all the three poly-herbal formulations were subjected to Ames II assay for ensuring their safety profile. Results epitomize that all the three poly-herbal formulations were devoid of significant mutagenic effect in TA98, and TAMix strains of Salmonella typhimurium under our experimental conditions.

Tinospora cordifolia, a safety evaluation.

Tinospora cordifolia is one of the indispensable medicinal plants used in veterinary folk medicine/Ayurvedic system of medicine for the treatment of diverse diseases and recommended for improving the immune system by means of body resistance. In the current study, we evaluated the genotoxic risk of the aqueous extract of T. cordifolia (TC) in a battery of four different genotoxicity tests viz., Ames, in vitro chromosome aberration (CA), rodent bone marrow micronucleus (MN), and Comet assay. Experimental results confirmed that in Ames test up to 5000 microg/plate of TC did not exhibit any mutagenic effect in Salmonella typhimurium mutant strains (TA97a, TA98, TA100, TA102, and TA1535). In CA assay, TC was not clastogenic to human peripheral blood lymphocytes up to a concentration of 3000 microg/ml. In MN and Comet assays, TC was pre-treated for 7 days at three dose levels (150, 200 and 250 mg/kg body weight) orally to male Balb/c mice. The results showed that TC treatment did not display clastogenicity and DNA damaging effect in bone marrow erythrocytes and peripheral blood lymphocytes respectively.

Evaluation of the genotoxic potential and acute oral toxicity of standardized extract of Andrographis paniculata (KalmCold).

Andrographis paniculata is used in the traditional medicine for cold and influenza remedy. The main endeavor in this study was to assess the genotoxicity of the standardized extract of A. paniculata (KalmCold) through three different in vitro tests: Ames, chromosome aberration (CA), and micronucleus (MN). Ames test was performed at 5000 microg/ml, 1581 microg/ml, 500 microg/ml, 158 microg/ml, 50 microg/ml, 16 microg/ml, while the clastogenicity tests were performed at 80 microg/ml, 26.6 microg/ml, 8.8 microg/ml for short-term treatment without S9; 345 microg/ml, 115 microg/ml, 38.3 microg/ml for short-term treatment with S9; and 46 microg/ml, 15.3 microg/ml, 5.1 microg/ml for long-term without S9 using DMSO as a vehicle control. Results of Ames test confirmed that KalmCold did not induce mutations both in the presence and absence of S9 in Salmonella typhimurium mutant strains TA98 and TAMix. In CA and MN, KalmCold did not induce clastogenicity in CHO-K1 cells in vitro. Based on our results, it is evident that KalmCold is genotoxically safe. Additionally in acute oral toxicity study, female rats were treated at 5000 mg/kg of KalmCold and observed for signs of toxicity for 14 days. KalmCold did not produce any treatment-related toxic effects in rats.