Evaluation of histo-toxicity of nimesulide in Black Kites (Milvus migrans): a pharmacodynamic study.

The present experimental work was conducted to elucidate the toxicity of nimesulide at three different doses in black kites (Milvus migrans). M. migrans is one of the most common raptors near human habitations. The goal of the current investigation was to determine whether nimesulide is similarly hazardous to raptors as was diclofenac sodium and to investigate the acute oral toxicity of nimesulide in these birds. For this study, eight adult male black kites (M. migrans) were randomly divided into four groups. M. migrans in the control group (n = 02) were not treated with nimesulide. The other three groups were given nimesulide doses. The birds in the first (n = 02) were declared the control group. The second (n = 02), third (n = 02), and fourth groups were administered nimesulide at a low, medium, and high dose of 2, 4, and 6 mg/kg live body weight of bird/day, respectively, for 10 days. Nimesulide-addled birds became listless and despondent, then anorexic. The birds were standing there with their eyes closed and showing no signs of life. There was an increase in saliva production, a slowing of breathing, and dilated pupils. No clinical signs were observed in the control group. No mortality was seen in the control or treated groups. The control group did not show lesions of gout, but black kites intoxicated with nimesulide at 2, 4, and 6 mg/kg live body weight of bird/day showed inflammation, apoptosis, hemorrhage, necrosis, and leukocytic infiltration tissues of the liver, kidney, and heart of black kites (M. migrans) treated with different concentrations of nimesulide. The treated groups also showed apoptosis of myofibrils and hyperplasia. The hypertrophy, atrophy, fibrosis, necrosis of skeletal muscles and hemorrhage were prominent in the muscles of black kites (M. migrans) intoxicated with nimesulide. All observed histological alterations got worse in a dose-related way. There was no significant difference in AST, ALT, ALP, serum uric acid, but a significant difference was observed in the values of serum urea (p = 0.001) and serum creatinine (p = 0.019).

Mosquitocidal activities of Chenopodium botrys whole plant n-hexane extract against Culex quinquefasciatus / Atividades mosquitocidas do extrato de n-hexano de planta inteira de Chenopodium botrys contra Culex quinquefasciatus

Abstract This research aimed to investigate various mosquitocidal activities of Chenopodium botrys whole- plant n-hexane extract against Culex quinquefasciatus. The extract showed remarkable larvicidal, pupicidal, adulticidal, oviposition deterrent and adult emergence inhibitory activities against Cx. quinquefasciatus. During the larvicidal and pupicidal activities, the 24-hour lethal concentration (LC50) of extract against 2nd instar larvae, 4th instar larvae and pupae were 324.6, 495.6 and 950.8 ppm, respectively. During the CDC (Centers for Disease Control and Prevention) bottle bioassay for adulticidal activity, the median knockdown times (KDT50) at 1.25% concentration was 123.4 minutes. During the filter paper impregnation bioassay for adulticidal activity, the KDT50 value at 0.138 mg/cm2 concentration was 48.6 minutes. The extract was fractionated into 14 fractions through silica gel column chromatography which were then combined into six fractions on the basis of similar retention factor (Rf) value. These fractions were screened for adulticidal activity by applying CDC bottle bioassay. The fraction obtained through 60:40 to 50:50% n-hexanes-chloroform mobile phase with 0.5 Rf value showed 100% adulticidal activity at 0.2% concentration. During oviposition deterrent activity, the highest concentration (1000 ppm) showed 71.3 ± 4.4% effective repellence and 0.6 ± 0.1 oviposition activity index. During adult emergence inhibition activity, the median emergence inhibition (EI50) value was 312.3 ppm. From the outcome of the present investigation, it is concluded that the n-hexane extract of C. botrys whole- plant possesses strong larvicidal, pupicidal, adulticidal, oviposition deterrent and adult emergence inhibitory activities against Cx. quinquefasciatus.

Resumo Esta pesquisa teve como objetivo investigar várias atividades mosquitocidas do extrato n-hexano de planta inteira de Chenopodium botrys contra Culex quinquefasciatus. O extrato mostrou atividades larvicida, pupicida, adulticida, dissuasora de oviposição e inibidora da emergência de adultos contra a Cx. quinquefasciatus. Durante as atividades larvicida e pupicida, a concentração letal de 24 horas (CL50) do extrato contra larvas de 2º estádio, larvas de 4º estádio e pupa foi de 324,6, 495,6 e 950,8 ppm, respectivamente. Durante o bioensaio com frasco do CDC (Centros para Controle e Prevenção de Doenças) para adulticida, o tempo médio de desativação (KDT50) na concentração de 1,25% foi de 123,4 minutos. Durante o bioensaio de impregnação com papel de filtro para a atividade adulticida do extrato, o valor KDT50 na concentração de 0,138 mg / cm2 foi de 48,6 minutos. O extrato foi fracionado em 14 frações através de cromatografia em coluna de gel de sílica que foram então combinadas em seis frações com base em um valor de fator de retenção (Rf) semelhante. Essas frações foram selecionadas quanto à atividade adulticida por meio da aplicação do bioensaio com garrafa do CDC. A fração obtida através da fase móvel de n-hexanos-clorofórmio 60:40% a 50:50% com valor de 0,5 Rf apresentou atividade adulticida de 100% na concentração de 0,2%. Durante a atividade de dissuasão da oviposição, a maior concentração de extrato (1000 ppm) apresentou repelência efetiva de 71,3 ± 4,4% e índice de atividade de oviposição de 0,6 ± 0,1. Durante a atividade de inibição da emergência de adultos, o valor médio de inibição da emergência (EI50) foi de 312,3 ppm. A partir do resultado da presente investigação, conclui-se que o extrato de n-hexano da planta inteira de C. botrys possui fortes atividades larvicida, pupicida, adulticida, dissuasora da oviposição e inibidora da emergência de adultos contra a Cx. quinquefasciatus.

Antidiabetic and hepato-renal protective effects of medicinal plants in STZ induced diabetic rats.

The antidiabetic and hepato-renal protective effects of Citrullus colocynthis and Momordica charantia ethanol extracts were investigated in streptozotocin (STZ) induced diabetic male albino rats. Diabetic rats were treated with C. colocynthis, M. charantia or C. colocynthis + M. charantia mixed extract at a dose of 250 mg /kg body weight per oral per day for 21 days. The mean body weight of all the diabetic rat groups on day 1 of treatment (day 10 of diabetes) was significantly lower than the normal control rat group (P<0.05). The blood glucose level of all the diabetic rat groups on day 1 of treatment (day 10 of diabetes) was significantly (P<0.05) higher (> 200 mg/dl) than the normal control rat group (95.5 ± 2.7). At the end of treatment (day 21), the diabetic rats treated with plant extracts showed significant increase (P<0.05) in body weight and significant (P<0.05) reduction in blood glucose level when compared to diabetic control animals. Significant increase (< 0.05) was observed in the serum bilirubin, alanine transaminase (ALT), aspartate transaminase (AST), alkaline phosphatase (ALP), urea and creatinine levels of diabetic control rat group. The serum levels of these liver and kidney-related parameters of diabetic rats treated with plant extract were significantly lower when compared to diabetic control rat group (p < 0.05). Photomicrographs of liver and kidney microsections from diabetic rats treated with these plant extracts showed amelioration in the hepato-renal histoarchitectures. It was concluded that the C. colocynthis and M. charantia methanol extracts are antidiabetic and hepato-renal protective in STZ induced diabetic male rats. Treatment of the diabetic rats with C. colocynthis + M. charantia mixed extract is more effective in the amelioration of diabetes and hepato-renal injuries in STZ induced diabetic male rats.

GCMS characterization and biological potential of the seeds and aerial part of Galium tricorne Stokes.

Natural products have long been proven very effective against various challenging diseases including cancer and bacterial infections. Galium tricorne is one of the important source of natural products, which has not been explored till date in spite of its profound ethnomedicinal prominence. The current study has been designed to explore the biological potential of G. tricorne and to extract and isolate chemical constituents from its aerial part and seeds respectively along with identification of their chemical constituents. Phytochemical screening was performed to figure out the presence of secondary metabolite in G. tricorne. Crude Methanolic extract (Gt.Crd), which was obtained from the aerial part while the fatty acids were extracted from the seeds, which were later on analyzed by GCMS. Similarly, Well Diffusion and MTT method were used for antibacterial activity and cancer cell line assay respectively. To evaluate the cytotoxic potential, brine shrimps were used. Likewise, in Gas Chromatography-Mass Spectroscopy (GC-MS) analysis a total number of 23 compounds were identified in Gt.Crd extract out of which 7 compounds were sorted out to have some sort of toxicity profile. In the same fashion, 5 fatty acids were identified in the seeds of G. tricorne. Moreover, among the fractions, chloroform fraction (Gt.Chf) exhibited greater zone of inhibition (ZOI) 20.37 mm followed by Gt.Crd 18.40 mm against S. aureus and S. pyogenes respectively. In cytotoxicity Gt.Chf was more active followed by ethyl acetate fraction (Gt.Eta) by exhibiting 88.32±0.62% (LC50=60 µg/mL) and 73.95±2.25% (LC50=80 µg/mL) respectively at 1000 µg/mL concentration of the tested sample. Gt.Chf exhibited greater cell line inhibitory activity (IC50=61 µg/mL) against HeLa cell line. Similarly, Gt.Crd displayed IC50 values of 167.84 µg/mL and 175.46 µg/mL against HeLa and NIH/3T3 cell line respectively. Based on the literature review and screenings, it may be concluded that the aerial part and seeds of G. tricorne are the rich sources of bioactive compounds. The results of the current study also authenticate the scientific background for the ethnomedicinal uses of G. tricorne.

Mosquitocidal activities of Chenopodium botrys whole plant n-hexane extract against Culex quinquefasciatus.

This research aimed to investigate various mosquitocidal activities of Chenopodium botrys whole- plant n-hexane extract against Culex quinquefasciatus. The extract showed remarkable larvicidal, pupicidal, adulticidal, oviposition deterrent and adult emergence inhibitory activities against Cx. quinquefasciatus. During the larvicidal and pupicidal activities, the 24-hour lethal concentration (LC50) of extract against 2nd instar larvae, 4th instar larvae and pupae were 324.6, 495.6 and 950.8 ppm, respectively. During the CDC (Centers for Disease Control and Prevention) bottle bioassay for adulticidal activity, the median knockdown times (KDT50) at 1.25% concentration was 123.4 minutes. During the filter paper impregnation bioassay for adulticidal activity, the KDT50 value at 0.138 mg/cm2 concentration was 48.6 minutes. The extract was fractionated into 14 fractions through silica gel column chromatography which were then combined into six fractions on the basis of similar retention factor (Rf) value. These fractions were screened for adulticidal activity by applying CDC bottle bioassay. The fraction obtained through 60:40 to 50:50% n-hexanes-chloroform mobile phase with 0.5 Rf value showed 100% adulticidal activity at 0.2% concentration. During oviposition deterrent activity, the highest concentration (1000 ppm) showed 71.3 ± 4.4% effective repellence and 0.6 ± 0.1 oviposition activity index. During adult emergence inhibition activity, the median emergence inhibition (EI50) value was 312.3 ppm. From the outcome of the present investigation, it is concluded that the n-hexane extract of C. botrys whole- plant possesses strong larvicidal, pupicidal, adulticidal, oviposition deterrent and adult emergence inhibitory activities against Cx. quinquefasciatus.

In-vitro and in-vivo evaluation of the antibacterial potential of Typha elephantina.

The present study was aimed to evaluate the in-vitro and in-vivo antibacterial effects of the Typha elephantina aqueous extract (TE.AQ), ethanolic extract (TE.ET) and T. elephantina methanolic extract (TE.ME) against eight selected clinical pathogens. The test samples were tested for in-vitro analysis (by disc diffusion method) at different concentrations of 5, 15, 25, 50 and 100 mg/dL against both gram positive and gram-negative strains. The highest potential was observed in TE.ME at a concentration of 100 mg/dL against Pseudomonas aeruginosa exhibiting 19.67 ± 0.577 mm zone of inhibition (ZOI). The same fraction also showed good activity against Staphylococus aureus with ZOI of 17.50 ± 0.70 mm. The TE.ET was found most active against P. aeruginosa and Streptococcus pyogenes having ZOI of 18.53 ± 0.503 and 16.2 ± 1.55 mm respectively at a concentration of 100 mg/dL. The most sensitive bacteria P. aeruginosa was selected for in-vivo study (using poultry chicks) for induction of infection in chicks. The effects of TE.AQ, TE.ET and TE.ME were determined at concentrations of 300 mg/kg body weight based on hematological parameters, liver enzymes and gross pathological findings of lungs and livers. The findings of the in-vivo study in chick's model showed that treatment of experimental animals with TE.ME significantly restored the hematological parameters, liver enzymes and architecture of lungs and livers. Based on scientific evidence, the current study suggests that TE.ME may serve as a best and new natural antibacterial agent and can be used against infections caused by P. aeruginosa.

Evaluation of wound healing property of Terminalia catappa on excision wound models in Wistar rats.

Wound is defined as the loss of breaking cellular and functional continuity of the living tissues. Management of wounds is frequently encountered with different problems. Drug resistance and toxicity hindered the development of synthetic antimicrobial agents with wound healing activity. Many plants with potent pharmacological activities may offer better treatment options viz. Terminalia chebula, Terminalia bellirica and Phyllanthus emblica formulations have shown healing activities on wounds.The present study was planned to investigate the wound healing activity of Terminalia catappa on excision wound model in rats. Ointment was prepared by using bark extract of Terminalia catappa in soft paraffin and preservative. Wistar albino rats (200-250 gm) of either sex were used in the present study. A circular wound of 2 cm in diameter was made on the depilated dorsal thoracic region of the rats under ether anesthesia in aseptic conditions. The ointment was applied for 18 days and percent wound closure observed along with the parameters viz. Epithelization, granuloma weight and scar formation. Animals were observed on 3rd, 6th, 9th, 12th, 15th and 18th post-wounding day.Wound healing activity was compared with that of control and Betadine ointment as standard drug. Animals treated with Terminalia catappa ointment exhibited 97% reduction in wound area as compared to the control animals (81%). Ointment treated wounds were found to induce epithelization faster compared to the control. In conclusion, Terminalia catappa ointment promotes significant wound healing in rats and further evaluation of this activity in humans is suggested.

Interaction of Al(2)O(3) nanoparticles with Escherichia coli and their cell envelope biomolecules.

AIMS: The aim of this study is to investigate the antibacterial activity of aluminium oxide nanoparticles (Al2 O3 NPs) against multidrug-resistant clinical isolates of Escherichia coli and their interaction with cell envelope biomolecules. METHODS AND RESULTS: Al2 O3 NPs were characterized by scanning electron microscope (SEM), high-resolution transmission electron microscope (HR-TEM) and X-ray diffraction (XRD) analyses. Antibacterial activity and interaction of Al2 O3 NPs with E. coli and its surface biomolecules were assessed by spectrophotometry, SEM, HR-TEM and attenuated total reflectance/Fourier transform infrared (ATR-FTIR). Of the 80 isolates tested, about 64 (80%) were found to be extended spectrum ß-lactamase (ESBL) positive and 16 (20%) were non-ESBL producers. Al2 O3 NPs at 1000 µg ml(-1) significantly inhibited the bacterial growth. SEM and HR-TEM analyses revealed the attachment of NPs to the surface of cell membrane and also their presence inside the cells due to formation of irregular-shaped pits and perforation on the surfaces of bacterial cells. The intracellular Al2 O3 NPs might have interacted with cellular biomolecules and caused adverse effects eventually triggering the cell death. ATR-FTIR studies suggested the interaction of lipopolysaccharide (LPS) and L-α-Phosphatidyl-ethanolamine (PE) with Al2 O3 NPs. Infrared (IR) spectral changes revealed that the LPS could bind to Al2 O3 NPs through hydrogen binding and ligand exchange. The Al2 O3 NPs-induced structural changes in phospholipids may lead to the loss of amphiphilic properties, destruction of the membrane and cell leaking. CONCLUSIONS: The penetration and accumulation of NPs inside the bacterial cell cause pit formation, perforation and disorganization and thus drastically disturb its proper function. The cell surface biomolecular changes revealed by ATR-FTIR spectra provide a better understanding of the cytotoxicity of Al2 O3 NPs. SIGNIFICANCE AND IMPACT OF THE STUDY: Al2 O3 NPs may serve as broad-spectrum bactericidal agents to control the emergent pathogens regardless of their drug-resistance mechanisms.

Botanicals to control soft rot bacteria of potato.

Extracts from eleven different plant species such as jute (Corchorus capsularis L.), cheerota (Swertia chiraita Ham.), chatim (Alstonia scholaris L.), mander (Erythrina variegata), bael (Aegle marmelos L.), marigold (Tagetes erecta), onion (Allium cepa), garlic (Allium sativum L.), neem (Azadiracta indica), lime (Citrus aurantifolia), and turmeric (Curcuma longa L.) were tested for antibacterial activity against potato soft rot bacteria, E. carotovora subsp. carotovora (Ecc) P-138, under in vitro and storage conditions. Previously, Ecc P-138 was identified as the most aggressive soft rot bacterium in Bangladeshi potatoes. Of the 11 different plant extracts, only extracts from dried jute leaves and cheerota significantly inhibited growth of Ecc P-138 in vitro. Finally, both plant extracts were tested to control the soft rot disease of potato tuber under storage conditions. In a 22-week storage condition, the treated potatoes were significantly more protected against the soft rot infection than those of untreated samples in terms of infection rate and weight loss. The jute leaf extracts showed more pronounced inhibitory effects on Ecc-138 growth both in in vitro and storage experiments.

Isolation, characterization, and identification of biological control agent for potato soft rot in Bangladesh.

A total of 91 isolates of probable antagonistic bacteria of potato soft rot bacterium Erwinia carotovora subsp. carotovora (Ecc) were extracted from rhizospheres and endophytes of various crop plants, different soil varieties, and atmospheres in the potato farming areas of Bangladesh. Antibacterial activity of the isolated probable antagonistic bacteria was tested in vitro against the previously identified most common and most virulent soft rot causing bacterial strain Ecc P-138. Only two isolates E-45 and E-65 significantly inhibited the in vitro growth of Ecc P-138. Physiological, biochemical, and carbon source utilization tests identified isolate E-65 as a member of the genus Bacillus and the isolate E-45 as Lactobacillus sp. The stronger antagonistic activity against Ecc P-138 was found in E-65 in vitro screening and storage potatoes. E-65 reduced the soft rot infection to 22-week storage potatoes of different varieties by 32.5-62.5% in model experiment, demonstrating its strong potential to be used as an effective biological control agent for the major pectolytic bacteria Ecc. The highest (62.5%) antagonistic effect of E-65 was observed in the Granola and the lowest (32.7%) of that was found in the Cardinal varieties of the Bangladeshi potatoes. The findings suggest that isolate E-65 could be exploited as a biocontrol agent for potato tubers.

Harnessing the purinergic receptor pathway to develop functional engineered cartilage constructs.

OBJECTIVE: Mechanical stimulation is a widely used method to enhance the formation and properties of tissue-engineered cartilage. While this approach can be highly successful, it may be more efficient and effective to harness the known underlying mechanotransduction pathways responsible. With this aim, the purpose of this study was to assess the effect of directly stimulating the purinergic receptor pathway through exogenous adenosine 5'-triphosphate (ATP) in absence of externally applied forces. METHODS: Isolated bovine articular chondrocytes were seeded in high density, 3D culture and supplemented with varying doses of ATP for up to 4 weeks. The effects on biosynthesis, extracellular matrix accumulation and mechanical properties were then evaluated. Experiments were also conducted to assess whether exogenous ATP elicited any undesirable effects, such as: inflammatory mediator release, matrix turn-over and mineralization. RESULTS: Supplementation with ATP had a profound effect on the growth and maturation of the developed tissue. Exogenous ATP (62.5-250 microM) increased biosynthesis by 80-120%, and when stimulated for a period of 4 weeks resulted in increased matrix accumulation (80% increase in collagen and 60% increase in proteoglycans) and improved mechanical properties (6.5-fold increase in indentation modulus). While exogenous ATP did not stimulate the release of inflammatory mediators or induce mineralization, high doses of ATP (250 microM) elicited a 2-fold increase in matrix metalloproteinase-13 expression suggesting the emergence of a catabolic response. CONCLUSIONS: Harnessing the ATP-purinergic receptor pathway is a highly effective approach to improve tissue formation and impart functional mechanical properties. However, the dose of ATP needs to be controlled as not to elicit a catabolic response.

Creating synergies for health systems strengthening through partnerships in Pakistan - a case study of the national eye health programme

Blindness and visual impairment are major causes of noncommunicable diseases in Pakistan. Two national population-based blindness surveys conducted in 1988 and 2002-04 demonstrated a reduction in prevalence of blindness from 1.78% to 0.9% with a significant drop in cataract blindness as a result of accelerated nationwide interventions and eye care integration in primary health care. In addition, between 2006 and 2008r 88 facilities were upgraded as a result of the national eye health programme. These measures resulted in a 279% increase in eye outpatient attendances and a 375% increase in eye surgeries performed. Investment in human resources development and policy change contributed significantly to the sustainability of the programme. Key challenges facing the programme include aligning national eye health strategies with health system strengthening informed through health systems research. This gaper attempts to document this extraordinary success

Effects of crude oil and diesel exposures on biochemical activities of polymorphonuclear leukocytes in cattle.

Cattle exposed to low doses of an Alberta crude oil, Pembina Cardium crude oil (PCCO), or a winter diesel oil no. 2 (WDO-2) were assessed for their biochemical activities in polymorphonuclear leukocyte (PMNL) cells (mainly neutrophils). The study used a randomized block design containing five treatment groups (8 animals/group). The animals were dosed per gavage with the test substance on study days 0, 14, 28, and 42. The dosages given (on per kg body weight) were: Group 1 (control), 10 mL/kg of potable water; Group 2, 5 mL/kg WDO-2; Group 3, 2.5 mL/kg PCCO; Group 4, 5 mL/kg PCCO; and Group 5, 10 mL/kg PCCO. Blood was collected at the specified intervals during the pre- and post-exposure periods, and the biochemical activities of isolated PMNL were analyzed. Cattle groups exposed to WDO-2 and PCCO showed moderate and statistically significant reductions (p < 0.01) in the activities of (1) phorbol myristate acetate (PMA) stimulated cellular respiration (respiratory burst), (2) NADPH-oxidase (PMA-stimulated production of superoxide anion), (3) myeloperoxidase, and (4) n-acetylglucosidase as compared to the control group. These biochemical parameters also showed statistically significant (p < 0.01) dose-related periodic (study day) trends. In general, these biochemical activities were decreased after each dosing; however, they subsequently recovered to near the pre-dosing levels. Such a biochemical response in PMNL provides a valuable biological tool to follow exposure effects in cattle accidentally exposed to low doses of petroleum hydrocarbons.

Pre-clinical assessment of DRF 4367, a novel COX-2 inhibitor: evaluation of pharmacokinetics, absolute oral bioavailability and metabolism in mice and comparative inter-species in vitro metabolism.

The aim of this study was to characterize the pharmacokinetics and determine the absolute bioavailability and metabolism of DRF 4367, a novel COX-2 inhibitor, in mice. In addition, the in vitro metabolism of DRF 4367 was studied in mouse, rat, dog, monkey and human liver microsomes. Following oral administration, maximum concentrations of DRF 4367 were achieved after about 1 h. Upon intravenous (IV) administration, the concentration of DRF 4367 declined in a bi-exponential fashion with a terminal elimination half-life of 4.0 h. The elimination half-life was unchanged with route of administration. The volume of distribution and systemic clearance of DRF 4367 in mice were 0.80 l h(-1) kg(-1) and 0.14 l kg(-1), respectively, after IV administration. The absolute oral bioavailability of DRF 4367 was 44%. In all species of liver microsomes examined, the primary route of metabolism for DRF 4367 was demethylation of benzyl methoxy to form a hydroxy metabolite (M1). The formation of this metabolite was mediated by CYP2D6 and CYP2C19 enzymes. M1 was not found to possess COX-2 inhibitory activity. Chemical-inhibition studies showed that quinidine (selective for CYP2D6) and ticlopidine (selective for CYP2C19) inhibited the formation of the hydroxy metabolite of DRF 4367, whereas potent inhibitors selective for other forms of CYP did not inhibit this oxidative reaction. Upon oral or IV administration of DRF 4367 to mice, unchanged DRF 4367, M1, the O-glucuronide conjugate of M1 (M1-G) and the O-sulfate conjugate of M1 (M1-S) were identified in bile.

Biochemical changes as early stage systemic biomarkers of petroleum hydrocarbon exposure in rats.

Crude and refined petroleum contain a complex mixture of aliphatic, aromatic, polyaromatic and heterocyclic hydrocarbon compounds. The objective of our research was to investigate early-stage biochemical changes in rats exposed to low dosages of petroleum hydrocarbons. The animals were repeatedly exposed, per oral by gavage, to low dosages (0.5-2.5 ml/kg) of an Alberta crude oil (ACO) and their general health and systemic biochemical parameters were assessed. Rats exposed to these doses of ACO did not show any apparent symptoms of intoxication. Similarly, no significant changes were observed in clinical parameters of systemic impairment. Systemic biochemical assessment has shown that ACO exposure caused marked changes in the activities of several cytochrome P-450 (CYP)-linked polysubstrate monooxygenase enzymes in liver, kidney and lung tissues. Exposure to ACO caused dose-dependent increases in the hepatic activities of ethoxyresorufin-O-deethylase, a CYP 1A1/A2-linked enzyme; pentoxyresorufin-O-dealkylase, a CYP 2B-linked enzyme, and ethoxycoumarin-O-deethylase, a CYP 2B/1A-linked enzyme. Temporal assessment showed that these systemic biochemical changes were reversible in nature. Analysis of biomarker chemicals provided evidence that in exposed animals petroleum hydrocarbons were mainly distributed in the adipose tissues.

Effects of multiple exposures of small doses of Pembina Cardium crude oil and diesel in rats.

In lands used for agricultural purposes, petroleum- or diesel-contaminated wastes and accidental spills of crude oil at some drilling sites pose exposure risks for occupational public, livestock, and wildlife. This study has assessed the effects of an Alberta crude oil, Pembina Cardium crude oil (PCCO), and a commercial diesel fuel #2 (CDF-2) in Sprague-Dawley rats after repeated exposures at small dose levels. Rats were given by gavage on day 1, 3, 5, and 8 specified dosages of either the control vehicle, methylcellulose (MC) (1.25 ml/kg), or PCCO (0.25-1.25 ml/kg), or CDF-2 (1.25 ml/kg). Exposure of rats to these dose levels of the test substances caused no overt symptoms of intoxication. A small but statistically significant increase in liver somatic index was observed in rats exposed to 1.25 ml/kg doses of PCCO and CDF-2; however, kidney somatic index was not significantly affected by these treatments. Blood analyses for hematological and clinical indicators of systemic impairments did not show any significant changes (p > 0.05) between the control and PCCO- or CDF-2-exposed rats. Biochemical assessment of liver and kidney tissues showed that compared to the control group, the PCCO- and CDF-2-exposed groups had a marked and significant increase (p < 0.05) in the hepatic activity of ethoxyresorufin-O-deethylase (EROD, a cytochrome P-450 [CYP] 1A1/A2-linked enzyme). In PCCO-exposed rats, the induction of EROD was dose-dependent. Exposure of rats with PCCO and CDF-2 also caused dose-related increases from the unexposed (control) or MC dosed rats in (1) hepatic activities of aryl hydrocarbon hydroxylase (AHH, a CYP 1A1-linked enzyme), ethoxycoumarin-O-deethylase (ECOD, a CYP 2B/1A-linked enzyme), glutathione transferase (GT), and NADPH-catalyzed microsomal lipid peroxidation; and (ii) ECOD activity in kidneys. The induction of hepatic CYP-linked enzymatic activities by PCCO and CDF-2 could be due to de novo synthesis of selected isoforms, as evidenced by the relative differences in the inhibition of EROD activity with 7,8-benzoflavone or metyrapone.

Purification, characterization, and molecular cloning of a chitinase from the seeds of Benincasa hispida.

A chitinase was purified from the seeds of Benincasa hispida, a medicinal plant also called white gourd, and a member of the Cucurbitaceae family. Purification was done by using a procedure consisting of only two fractionation steps: an acid denaturation step followed by ion-exchange chromatography. The sequence of the N-terminal forty amino acid residues was analyzed and the sequence indicated that the enzyme is a class III chitinase. The enzyme, which is a basic chitinase, is one of at least five chitinases detected in the seed extract of B. hispida. Like other class III chitinases, this enzyme also has lysozyme activity. A genomic clone of the gene encoding the enzyme was isolated and sequenced. The gene has the potential to encode a protein of 301 amino acid residues. The deduced amino acid sequence of the protein, as expected from the N-terminal amino acid sequence, shares high degrees of similarity with other class III chitinases.

Purification of an osmotin-like protein from the seeds of Benincasa hispida and cloning of the gene encoding this protein.

A pathogenesis-related (PR) protein was purified from the seeds of Benincasa hispida, which is a medicinal plant and a member of the Cucurbitaceae family. Purification was achieved by using a procedure consisting of an acid treatment step followed by two chromatography steps. The protein is a basic protein with molecular mass of approximately 28 kDa. The sequences of the N-terminal 30 amino acids and four peptides generated from protease digestion were determined. These sequences indicated that the protein is an osmotin-like protein (OLP). Osmotin and OLPs are members of the thaumatin-like, PR-5 family of the PR proteins. A genomic clone of the gene encoding the protein was isolated and sequenced. The predicted protein has a signal peptide of 18 amino acids, and the mature protein has a molecular mass of 24.8 kDa with an isoelectric point of 7.67. The protein has 17 cysteine residues, of which 16 appear in the same positions as those appear in the sweet-tasting protein thaumatin and several other thaumatin-like proteins. Southern hybridization analysis indicated that the gene encoding the protein is a single copy gene. A computer-generated, three-dimensional model of the protein is presented.

Protection against lipopolysaccharide-induced death by fluoroquinolones.

Because fluoroquinolones have an immunomodulatory effect on cytokine production by lipopolysaccharide (LPS)-treated human monocytes, we examined the effect of fluoroquinolones on the survival of mice injected with a lethal dose of LPS. Trovafloxacin (100 mg/kg), ciprofloxacin (250 mg/kg), and tosufloxacin (100 mg/kg) protected 75% (P = 0.0001), 25% (P = 0.002), and 50% (P = 0.002), respectively, of mice against death. The fluoroquinolones significantly reduced serum levels of interleukin-6 and tumor necrosis factor alpha in LPS-treated mice. The protective effects of fluoroquinolones in LPS-induced shock in mice may also occur in humans.

L-carnitine moderately improves the exercise tolerance in chronic stable angina.

OBJECTIVES: To study the safety and efficacy (with reference to exercise ECG testing) of oral L-carnitine in chronic stable angina. METHODS: Forty-seven patients, 30 men and 17 women, aged 56 +/- 8 years, were randomized to receive L-carnitine (n = 28) or placebo (n = 19) in the dose of 2 g/day for 3 months. The adjuvant treatment was not changed during the study. Patients were evaluated by computerized stress test (CST) done at the beginning and end of the trial. The parameters assessed were exercise duration, time to onset of ST changes, total ST score at peak exercise, rate-pressure product at peak exercise, and time needed for the ST changes to recover to baseline. RESULTS: The two groups were comparable at the beginning of the study. There was no change in the CST parameters in the placebo group at the end of 3 months. In the L-carnitine group there was a statistically significant improvement in the exercise duration from 7.8 +/- 2.2 min to 8.6 +/- 1.8 min (p = 0.006) and in the time needed for the ST changes to revert to baseline from 7.2 +/- 3.9 min to 5.7 +/- 3.8 min (p = 0.019). No change was noted in the time to onset for ST depression, ST score and double product. There were no systemic adverse effects or coronary events in either group. CONCLUSION: Oral L-carnitine is safe and moderately improves the duration of exercise and time to recovery of ST changes in patients with chronic stable angina.