Pharmacokinetics and in vivo chemosuppressive activity studies on cryptolepine hydrochloride and cryptolepine hydrochloride-loaded gelatine nanoformulation designed for parenteral administration for the treatment of malaria.

The main objective of this investigation was to establish the pharmacokinetics profile and in vivo chemosuppressive activities of cryptolepine hydrochloride-loaded gelatine nanoparticles (CHN) designed for parenteral administration for the treatment of malaria in comparison to the drug free in solution (CHS). Single-dose pharmacokinetics was investigated in Wistar rats by administering CHN or CHS (equivalent to 10 mg/kg of drug) by IV bolus injection via the lateral tail vein. The drug concentration in plasma was monitored over a 24-h period following administration. Chemosuppressive activity was investigated in Wistar rats challenged with P berghei parasites. Animals were given a daily dose of either CHN or CHS, equivalent to 2.5-100 mg/kg by intraperitoneal injection. The level of parasitaemia was determined by light microscopy by examining Giemsa-stained thin blood smears prepared from the tail end on day four of infection. It was found that CHN attained a higher (4.5-folds) area under the curve (AUC (0-24)) compared to CHS. CHS however produced a higher volume of distribution (4-folds). Distribution and elimination rates were higher with CHS which resulted in a lower (11.7 h) elimination half-life compared to that of CHN (21.85 h). The superior pharmacokinetic profile of CHN translated into superior chemosuppressive activity at all dose levels relative to CHS. As a conclusion, loading cryptolepine hydrochloride into gelatine nanoparticles improved both pharmacokinetics and in vivo antiplasmodial activity of the compound with the highest chemosuppression (97.89 ± 3.10) produced by 100 mg/kg of CHN.

Genotoxicity, cytotoxicity and toxicological evaluation of whole plant extracts of the medicinal plant Phyllanthus niruri (Phyllanthaceae).

Phyllanthus niruri is a medicinal plant (commonly known as stone breaker) found in the tropics and other parts of the world. It is known for its capacity to block the formation of calcium oxalate crystals and kidney stone formation in urolithiasis. This plant has been used to treat hyperglycemia, hypertension, pain, and mild cases of malaria. We examined the geno-, cyto- and overall toxicity of P. niruri whole plant ethanolic extract. The extract was administered as a single dose of 30 or 300 mg/kg to laboratory rats by gavage, accompanied by negative (0.9% saline) and positive (10 mg/mL N-ethyl-N-nitrosourea) controls that were injected intramuscularly 48 h after extract administration. The ratio of polychromatic (PCE)/normochromatic erythrocytes (NCE) from femur bone marrow was scored for genotoxicity. Cytotoxicity was determined using descending concentrations (0.2-0.0125 g/mL) of the extract incubated with peripheral blood mononuclear cells. Lactate dehydrogenase release from damaged cells was determined and the CC(50) calculated. Subchronic administration of the extract at 30 or 300 mg/kg was done for 90 days to determine general toxicity. PCE:NCE (%) for the extract and negative control was 63, compared to 168 (positive control). The CC(50) was 26.3 mg/mL and hepato-renal toxicity after subchronic extract administration was nil. We conclude that ethanol extract of P. niruri is not cytotoxic or genotoxic, and is generally non-toxic on subchronic administration.

Menarcheal and pubertal development and determining factors among schoolgirls in Kumasi, Ghana.

To determine menarcheal and pubertal ages and possible factors responsible for current pubertal trends in Kumasi, Ghana, a cross-sectional study was conducted among 720 urban and rural Kumasi 7-17-year-old schoolgirls and their mothers in 2008. Heights and weights were measured and additional information obtained by survey. Mantel-Haenzsel, χ(2), ordered logistic regression and probit analyses were used to analyse the data collected. With 40.42% menarcheal prevalence, median menarcheal ages were significantly different: 12.37 ± 1.48 years urban and 13.41 ± 2.25 years rural; 12.89 ± 1.93 years, overall. Obesity (OR = 2.57; p = 0.033) and high socioeconomic status (OR = 2.12; p = 0.008) were predictors of early menarche, while a younger mother was protective against early menarche (OR = 0.32; p = 0.039). Age at menarche among Kumasi schoolgirls has dropped 0.76 years since it was last determined among similarly aged girls in 1986, declining at a rate of about 0.32 years/decade. The predicting factors provide an important opportunity for intervention through school curricula and targeted education of adolescents.

Degradation of aflatoxin B(1) by cell-free extracts of Rhodococcus erythropolis and Mycobacterium fluoranthenivorans sp. nov. DSM44556(T).

Biological degradation of aflatoxin B(1) (AFB(1)) by Rhodococcus erythropolis was examined in liquid cultures and in cell-free extracts. Dramatic reduction of AFB(1) was observed during incubation in the presence of R. erythropolis cells (17% residual AFB(1) after 48 h and only 3-6% residual AFB(1) after 72 h). Cell-free extracts of four bacterial strains, R. erythropolis DSM 14,303, Nocardia corynebacterioides DSM 12,676, N. corynebacterioides DSM 20,151, and Mycobacterium fluoranthenivorans sp. nov. DSM 44,556(T) were produced by disrupting cells in a French pressure cell. The ability of crude cell-free extracts to degrade AFB(1) was studied under different incubation conditions. Aflatoxin B(1) was effectively degraded by cell free extracts of all four bacterial strains. N. corynebacterioides DSM 12,676 (formerly erroneously classified as Flavobacterium aurantiacum) showed the lowest degradation ability (60%) after 24 h, while >90% degradation was observed with N. corynebacterioides DSM 20,151 over the same time. R. erythropolis and M. fluoranthenivorans sp. nov. DSM 44,556(T) have shown more than 90% degradation of AFB(1) within 4 h at 30 degrees C, whilst after 8 h AFB(1) was practicably not detectable. The high degradation rate and wide temperature range for degradation by R. erythropolis DSM 14,303 and M. fluoranthenivorans sp. nov. DSM 44,556(T) indicate potential for application in food and feed processing.

Subchronic toxicity studies of the antidiabetic herbal preparation ADD-199 in the rat: absence of organ toxicity and modulation of cytochrome P450.

The subchronic toxicity of the aqueous antidiabetic herbal extract ADD-199, prepared from Maytenus senegalensis, Annona senegalensis, Kigelia africana and Lanneawelwitschii, and administered at a daily dose of 100 or 500 mg/kg body weight over 30 days, was investigated in male Wistar albino rats. Certain haematological, urine and plasma biochemical parameters, and modulation of some hepatic cytochrome P450 (CYP) isozymes were measured as indices of organ specific toxicity or potential for drug interactions. ADD-199 did not affect plasma aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP) and albumin or creatinine kinase (CK) levels. It also did not affect plasma creatinine and urea levels. Furthermore, ADD-199 neither affected PCV nor blood Hb, RBC, reticulocytes, platelets, lymphocytes and granulocyte levels. It, however, caused significant dose-dependent reductions in WBC counts at day 15 with varying degrees of recovery by day 30. It also reduced the rate of body weight increases after week 3. However, no changes were observed in organ weights at termination. ADD-199 did not significantly affect zoxazolamine-induced paralysis and pentobarbital-induced sleeping times as well as certain CYP isozyme activities in rats. These findings suggest that ADD-199 had no overt organ specific toxicity and did not demonstrate a potential for drug interactions via CYP-mediated metabolism in the rat on subchronic administration.

Findings from a buruli ulcer mouse model study.

SUMMARY INTRODUCTION: Buruli ulcer disease is endemic in many developing countries in Africa. It is caused by Mycobacterium ulcerans, a toxin-producing bacterium with predilection for the skin and its deeper tissues. The exact mode of transmission is unclear and the pathogenesis is also not well understood, necessitating further elucidation through animal studies. OBJECTIVE: The study assessed the infectivity of a Ghanaian Mycobacterium ulcerans isolate and the dose-response pattern in BALB/c mice. METHOD: Ten standardized bacterial suspensions of different concentrations were prepared from the M. ulcerans isolate and inoculated into the foot-pads of the mice. Thereafter they were observed for clinical signs of Buruli ulcer, upon which they were serially euthanised and evaluated for pathological and microbiological changes. RESULTS: Irrespective of the inoculum dose, all the experimentally infected mice developed similar clinical lesions, from erythema to foot ulceration (3.1 to 6.7 weeks after inoculation). However, the higher the inoculum dose the earlier the onset of the lesions. After the development of foot ulceration, mice that had received between 1 to 4 doses developed gangrene (5.7 to 7.2 weeks after inoculation) and died within a week, while those that had received 5 to 10 doses lost their limbs spontaneously (5.6 to 6.1 weeks after inoculation), followed by sudden clinical recovery. Eight weeks after the spontaneous amputation the amputees relapsed with concomitant metastasis, anasarca and death. Acid-fast bacilli (AFBs) were detected in inoculated and non-inoculated limbs, tails, visceral organs, faecal pellets and caecal contents of the mice. The AFBs detected in the caecal samples were innumerable and unusually long. Though AFBs were consistently detected in lymph nodes they were never detected in blood samples. CONCLUSION: The findings suggest that the progression and final outcome of an M. ulcerans infection maybe dose related. The unequivocal absence of AFBs in the blood, but their consistent presence in lymph nodes located in the lower limbs right up to the neck, suggests that the microbes are disseminated through the lymphatic system rather than through the blood.

Enhanced depletion of glutathione and increased liver oxidative damage in aflatoxin-fed mice infected with Plasmodium berghei.

The effect of dietary aflatoxins B1 and G1 and Plasmodium berghei infection on glutathione (GSH) levels and liver status in mice was investigated. Three days after intraperitoneal injection of 0.1 x 10(6) parasitized red blood cells into the mice, there was a significant fall in blood glutathione levels accompanied by a significant increase in serum cholinesterase and liver malonic dialdehyde levels in the mice fed aflatoxin compared with those in the control group. The results suggested that malaria parasites can enhance depletion of host glutathione and oxidative damage of the liver in mice fed low levels of aflatoxins.

Comparative effects of aflatoxins G1 and B1 at levels within human exposure limits on mouse liver and kidney.

ddy mice were exposed to aflatoxins B1 and G1 via their feed (4.8 ng AFG1, 0.8 ng AFB1 or both/kg body wt./day) while in utero. At six months of age, hepatorenal studies were carried out. The AFG1 caused significant accumulation of only neutral fat in the liver, a slight rise in serum triglyceride and intensified hepatorenal inflammation, necrosis and bile duct proliferation. The AFB1, caused the accumulation of both neutral fat and fatty acids in the liver, and was cytotoxic to the liver and kidney. Iron storage of the liver, hematological indices, serum total protein and albumin levels were not affected by the aflatoxins.

Isolation of Corynebacterium renale from slaughtered cattle at the Zaria abattoir in Nigeria.

Forty urinary bladders were collected from apparently healthy cattle slaughtered at the Zaria abattoir. Twenty-four (60%) were from male animals and 16 (40%) from females. Urine samples were obtained and cultured for Corynebacterium renale. Four (16.7%) of the samples from males and 1 (6.3%) from females were positive. The difference in infection between the sexes was statistically significant (P less than 0.001).

Serological and cultural examination for human leptospirosis in Plateau State, Nigeria.

In a serological examination of 710 serum samples collected from human volunteers in Plateau State, Nigeria, 128 (18.0pc) had leptospiral antibody titres of 1:100 and above. The prevalence of antibodies to individual serovars were: hardjo 28 (21.9pc), pomona 18 (14.1pc), canicola 17 (13.3pc), grippotyphosa 15 (11.7pc), pyrogenes 13 (10.2pc), icterohaemorrhagiae 12 (0.4pc) and autumnalis 8 (6.3pc). There was no statistical difference in the prevalence rate of leptospirosis in the different local government areas (p greater than 0.05; X2). Among the occupational groups examined, the abattoir workers were particularly at risk with a prevalence rate of 29.5pc. A leptospiral variant of strain Hardjoprajitno was also isolated from the midstream urine of an abattoir worker who was apparently healthy. The isolation of Leptospira interrogans serovar hardjo in man is the first such report in Nigeria.

A new leptospiral serovar in the Pyrogenes serogroup isolated in Nigeria.

Five leptospiral strains were isolated from bovine kidneys during a cultural survey for pathogenic leptospires in Nigeria. Preliminary test results indicated that the five strains were identical and serologically heterologous to the other members of the Pyrogenes serogroup. Further examination of the strains by the cross-agglutinin absorption test, factor analysis and restriction endonuclease analysis confirmed that the strains constitute a new serovar. It is therefore proposed that this strain be recognised and designated as serovar nigeria, type strain Vom.

Serological prevalence of bovine leptospirosis in Plateau State, Nigeria.

Serum samples obtained from 1.537 cattle in the 14 local government areas (LGAs) of Plateau State of Nigeria were screened for the presence of leptospiral antibodies using 13 serovars in a modified microscopic agglutination test (MAT). Two hundred and twenty-two (14.4 p.100) of the cattle tested had leptospiral antibody titres of 1:100 or higher to one or more of the test antigens. The prevalence rates of antibodies to individual serovars were: hardjo (35.6 p.100), pomona (11.7 p.100), pyrogenes (11.7 p.100), canicola (9.5 p.100), grippotyphosa (7.7 p.100), bratislava (5.9 p.100), icterohaemorrhagiae (5.9 p.100), ballum (4.5 p.100), autumnalis (3.6 p.100), bataviae (2.3 p.100) and tarassovi (1.8 p.100). The serological prevalence of bovine leptospirosis in the various local government areas of Plateau State of Nigeria differed significantly (P less than 0.05; X2).

Vaccination challenge studies with variants of street rabies virus isolated in Nigeria.

In a preliminary study it was observed that adult ICR mice immunized with serial dilutions of an inactivated experimental human rabies vaccine from the Pitman-Moore (PM) vaccine virus were well protected against challenge with homologous virulent PM virus and challenge virus standard (CVS). However only one of five variant representatives in five of seven groups of 41 isolates of street rabies virus from Nigeria characterized by hybridoma monoclonal antibodies specific for the nucleocapsid and glycoprotein antigens of rabies virion was protected for by the vaccine. Guinea pigs immunized with a live attenuated low egg passage (LEP, Flury strain) vaccine currently used in canine vaccination in Nigeria protected against challenge with all five variants. The LEP vaccine protected against the variants and CVS quite well even when 1:125 dilution of the vaccine was used.

Atypical Pasteurella haemolytica type A from poultry.

Atypical strains of Pasteurella haemolytica that failed to ferment maltose were isolated from nodular necrosis in the liver and heart blood of domestic fowl (Gallus domestica). These strains did not typically behave like either of the two well-known biotypes of P. haemolytica. The strains utilized trehalose and produced hydrogen sulfide (H2S), thus behaving like P. haemolytica type T, and produced acid in xylose but not in salicin, thus behaving like P. haemolytica type A. Most of the properties of the strains, however, conformed closely to those of P. haemolytica type A. Detailed characteristics of the isolates are described and discussed.

Bacteriological investigation of the postpartum uterus: Relationship to involution and histopathological findings.

A bacteriological investigation of the postpartum uteri of suckled and nonsuckled Bunaji (White Fulani) cows was undertaken to determine the microflora, the effect of suckling on the flora and the influence of the flora on uterine involution and histopathology. Uterine contamination by bacteria was highest between days 10 and 21 postpartum with Staphylococcus aureus being the commonest organism isolated. There was no consistent isolation of one type of bacterial organism from a cow throughout the sampling period. Suckling did not affect the incidence of uterine contamination by bacteria. Foci of leucocytes (mainly lymphocytes) were found in uterine sections in an increasing frequency with days postpartum. Bacteria were isolated from only 3 of the 12 uteri that had the leucocytic foci. Uterine involution was normal and complete by 25 days after calving irrespective of the presence or absence of bacteria or leucocytic foci. We concluded that postpartum uterine contamination by bacteria was not influenced by suckling and that puerperal uterine contamination by bacteria did not interfere with the normal process and duration of uterine involution.