Asian Pacific Journal of Tropical Medicine

1-7

Authors: Jangra Pooja, Singh Ajit

Objective: To isolate and characterize Staphylococcus aureus (S. aureus) β-hemolysinneutralizing dAbs from phage display library of Indian desert camel. Methods: Phage display library of 5×107 dAb clones of LPS-immunized Indian desert camel constructed in our laboratory was used for selection of S. aureus exotoxin-specific clones by panning technique. Enrichment of Ag-specific clones in successive rounds of panning was assessed by phage-ELISA and phage titration. Different dAb clones binding to S. aureus exotoxin Ags were expressed with C-terminal 6×His tag in E. coli and purified by Ni-chelate chromatography. The expression was verified by SDS-PAGE and western blotting. The purified clones were tested for inhibition of 'hot-cold' hemolytic activity in vitro. Resistance to thermal inactivation of the dAb clones was studied by observing the effect of heat treatment from 50 ℃ to 99 ℃ for 30 mins on the 'hot-cold' hemolytic activity in vitro. Results: Several dAb clones binding to S. aureus exotoxins were isolated and enriched by three rounds of panning. The soluble dAb clones were approximately ~16 kDa in size and reacted with 6×His tag specific murine monoclonal antibody in western blot. One of the Ni-chelate affinity purified dAb.6×His clones, inhibited S. aureus β-hemolysin activity in vitro and resisted thermal inactivation upto 99 ℃. Conclusions: An S. aureus β-hemolysinneutralizing dAb clone of possible therapeutic potential has been isolated.

13-16

Authors: Ukwe Chinwe Victoria, Ubaka Chukwuemeka Michael, Madusque Uche Johnny

Objective To evaluate the antiulcer activity of the methanolic root extract of the plant Olax subscorpioidea in experimental rats. Methods Phytochemical tests and acute toxicity tests were carried out on its methanolic root extract. Pre-treatments with three doses of the extract (200, 400 and 600 mg/kg body weight orally) and Sucralfate at 100 mg/kg orally were used for the various groups of rats. The indomethacin and ethanol models for experimental induction of ulcers in rats were used. Mean ulcer indices were measured and percentage inhibition was derived. Results Phytochemistry revealed presence of alkaloids, steroids, glycosides and terpenoids and the extract showed an LD50 of 2 154 mg/kg in mice. Ulcer was produced in all the rats in both models with the extract showing potent antiulcer activity of ethanol model. There was no significant ulcer inhibition by any of the treatments compared to control group in the indomethacin model but the extract's antiulcer effect was dose-dependent (11.8%, 19.2%, 32.7%, P >0.05). The ulcer reduction in the ethanol model was significant ulcer reduction in the highest dose group and Sucralfate group compared to control group (79.3% and 82.9%, P < 0.05). However, the extract at all dosage showed a dose-dependent ulcer inhibition in this model. Conclusion The above results suggest that the roots of Olax subscorpioidea possess antiulcer activity in experimental rats as claimed by traditional users.

21-24

Authors: AW Obianime, FI Uche

Objective To investigate the effects of the aqueous extracts of the seeds of Tetracarpidium conophorum and the effect of Proviron (12.5 mg/kg) (as standard) on the hormonal parameters of male guinea pigs, compare the effects of the seeds of Tetracarpidium conophorumand Proviron, and screen the phytochemical constituents of the seeds of Tetracarpidium conophorum. Methods The hormonal effects of the Tetracarpidium conophorum and Proviron were tested by hormonal assay, using enzyme immuno assay method. This was done by reaction of antibody with serum testosterone and testosterone label, magnetic solid phase separation and colour development step. Phytochemical screening was done using standard procedures. Results The aqueous extract of the Tetracarpidium conophorum seeds (100–400 mg/kg) caused a statistically significant increase (P < 0.05, ANOVA) in the level of testosterone of male guinea pigs, from (2.60 ± 0.06) ng/mL to (3.40 ± 0.05) ng/mL, (3.00 ± 0.60) ng/mL and (3.30 ± 0.45) ng/mL on the 7th, 14th and 21st day of the administration of the extracts, respectively. The highest increase was obtained after the 7th day of treatment [(3.40 ± 0.05) ng/mL]. These effects were very comparable to the effects of Proviron on the testosterone of male guinea pigs, which were obtained to be (2.80 ± 0.01) ng/mL, (2.90 ± 0.16) ng/mL and (3.10 ± 0.30) ng/mL on the 7th, 14th and 21st day, respectively. These effects were dose-and time-dependent. The optimum effect on testosterone level under dose-dependent study [(4.70 ± 0.45) ng/mL] was obtained at 300 mg/kg of Tetracarpidium conophorum after 7 days treatment. Finally, the phytochemical screening of the seeds of Tetracarpidium conophorum revealed the presence of flavonoids, tannin, alkaloids, carbohydrate, volatile oils, terpenoids, saponins and cardiac glycosides. Conclusion This study supports the claims on the use of the seeds of this plant by traditional medicine practitioners as a fertility agent. However, further studies need to be done to isolate and characterize the active principle(s) responsible for this activity in this plant.

29-31

Authors: Reginald NP Nwankwoala, OA Georgewill

Objective To investigate the activity of the active principle in the spines of the seed pods of Mucuna pruriens using contraction of guinea pig ileum as index of pharmacological activity. Methods The active principle was extracted with 0.0015 M NaCl. Muscle strips of guinea pig ileum were prepared and contractile responses were measured using a Kymograph. Two sets of experiment were conducted: (1). The contraction of the ileum in presence of different concentrations of histamine, 2-methylhistamine and the extract of Mucuna pruriens. (2). The contractile response of the ileum in presence of different concentrations of the extract and antagonists including diphenhydramine, atropine and methysergide. Results (1) The extract of Mucuna pruriens hair, 2-methylhistamine and histamine produced dose dependent contraction of guinea pig ileum (Extract ED50 = 13.0 μg/mL, 2-methylhistamine ED50 = 8.5 μg/mL and histamine ED50 = 10.0 μg/mL). (2) Diphenhydramine, an H1 antagonist competitively blocked the contractile response of the Mucuna pruriens extract. (3) Coadminstration of the Mucuna pruriens extract either with different doses of antimuscarinic agent atropine or 5-hydroxytryptamine blocking agent methysergide did not alter the extract induced contractile response of the guinea pig ileum. Conclusion These results demonstrate that the spines of Mucuna pruriens possess histamine activity which may contribute to its itching and painful irritation effects.

36-40

Authors: A Shashi, N Sharma, M Bhardwaj

Objective To monitor the pathological alterations in pancreas of rat during experimental fluorosis. Methods Sixty Sprague Dawley albino rats of both sexes were divided into 12 experimental groups and one control group. The rats of control group were administered subcutaneously double distilled water 1 mL/kg bw daily. The experimental groups were injected with 30, 45, and 75 mg NaF/kg bw/day. The experimental period was divided into 4 phases at interval of 15, 22, 30, and 36 days. Animals were sacrificed from each group at the end of 16, 23, 31, and 37 days. Results The following changes were observed in this study: (1) Pathological examination of pancreas after 15 days of fluoride treatment revealed: hypertrophy of acini, leucocytes infilteration and pycnotic nuclei due to necrosis of acini in group 1; uremic alterations, invulsion and infoldings of reticular layer of islets of Langerhans in group 2; and a decrease in number of acini and interlobular connective tissues resulted in an increase in intercellular spaces in pancreas of rats in group 3. (2)Hyalinization and hypertrophy in the lobules of acini and hyperplasia and hypertrophy in intercalated duct with mucinous secretion in pancreas of rat of group 4; squamous metaplasia of pancreatic duct, adenoma of pancreas, hemorrhagic necrosis in group 5; and hyperplasia of acini and reduction in number of pancreatic islets in group 6. (3) Disorganization and atrophy of pancreatic lobules and presence of vacuoles in a group of six were visible in pancreas of rats in group 7; acute pancreatic and lamellated inflammatory cells in test rats of group 8; and islet adenoma and decrease in number of islets cells, and exudation in acini were noticed in experimental rats of group 9. (4) In the last phase of experimentation, atrophic alterations in pancreatic acini, invulsions, and necrosis was prominent in group 10, deep inflammation and proliferation of connective tissue of pancreas in experimental group 11, and periodical fibrosis, hyperplasia of acini, degenerative changes in pancreas of rats in group 12. Conclusion The histopathological examination of pancreas of fluoridated rats exhibited structural alterations in the exocrine glands. The acini revealed hypertrophy, pyknotic nucleus, necrosis and uremic alterations. Acini became lobulated and reveal increased pigmentation.

45-47

Authors: Mohammad Rahbar, Mohsen Zahraei, Aboulgasm Omidvarnia, Mohammad Taqi Afshani, Mostafa Glami, Rogaeieh Sabourian, Shahla Farsi, Hosein gholami, Saeid Mahdavi, Parisa Islami, Mona Mohammad Zadeh, Massoud Hajia, Azar Valipour, Rana Amini

Objective To determine epidemiology and antimicrobial susceptibility patterns of Vibrio (V.) cholerae O1 biotype EL Tor in summer outbreak of 2008 in Iran. Methods Stool samples were collected from patients suspected to have cholera admitted to hospitals and clinics. Specimens examined by conventional bacteriological methods. All isolates were sent to cholera reference laboratory for further confirmation, stereotyping and susceptibility testing. Results A total of 220 patients were diagnosed as cholera. All cases confirmed by Iranian reference health laboratory. One hundred ninety nine of 220 V. cholerae serotypes were Inaba and 21 serotypes were Ogawa. All cases were reported from thirteen provinces. The majorities of cases were from Tehran, Qum and Zahedan provinces with 56, 26 and 25 cases respectively. 24(11%) of patients were under 15 years old and 196 (89%) of patients were older than 15 years.149 (68%) of patients were male and 71 (32 %) were female. 129(59%) of patients had Iranian nationality,79 (36.5%) were from Afghanistan and, 12 (5%) were from Pakistan. All isolates were resistant to co-trimoxazole, nalidixic acid, furazolidone, and intermediate to chloramphenicol and were susceptible to tetracycline, ciprofloxacin, and erythromycin. Conclusion Our study reveals that in recent outbreak caused by V. cholerae EL Tor serotype Inaba is the predominant serotype. All isolates are resistant to cotrimoxazole, nalidix acid and furazolidon.

51-54

Authors: TU Agan, JE Ekabua, CU Iklaki, A Oyo–Ita, I Ibanga

Objective To evaluate factors associated with prevalence of malaria parasitaemia at first antenatal care visit. Methods The study was conducted at the University of Calabar Teaching Hospital from 1st June, 2007 to 31st July, 2007. A structured questionnaire was administered to a total of 545 pregnant women that were recruited in this study after obtaining informed consent and two slides of thin and thick films were prepared for each participant. Results Five hundred and twenty (95.4%) out of the 545 participants suffered from malaria parasitaemia, the rest 4.6% of those who had no parasitaemia had experienced symptomatic malaria before and were treated in private hospitals prior to their recruitment into the study. All participants (100%) who did not have antimalarials had parasitaemia compared with 91.1% among those that had antimalarials. The proportion of moderate to severe parasitaemia was also significantly higher among the former. Besides, the difference in parasitaemia between primigravidae and multigravidae was statistically significant (P=0.000) too. Among the methods used for vector control, only insecticide treated nets (ITNS) was associated with significant reduction in the level of parasitaemia (RR =0.83). Conclusion Malaria parasitaemia at first booking is significantly higher in primigravidae and women who have no anti-malaria treatment. The use of safe and effective antimalarial treatment along with ITNs will significantly reduce the level of parasitaemia in pregnant women.

59-62

Authors: S Fayed, S Abd El Dayem, H Hussein, M Sherief, S El–Naghi

Objective To detect the prevalence of Helicobacter polori (H. pylori) in a group of pediatric patients complaining of gastrointestinal disorders, and to identify the virulent strains of H. pylori and its relation to peptic disease. Methods The study included 60 patients (41 males and 19 females), aged from 6 months to 14 years, with complaining of different gastrointestinal troubles. Anti-H. pylori IgG antibodies were used. Thorough examination of the oesophagus, stomach and duodenum was done in all patients using a fibreoptic endoscopy. In addition, gastric antral biopsies were taken for diagnosis of H. pylori infection in the following test: rapid urease test, histopathologic examination and culture. And cytotoxin assay of virulent strains was employed. H. pylori infection was diagnosed if 3 or more tests for diagnosis were found to be positive. Results 33 out of 60 patients (55%) was positive. 12 (20%) patients had positive toxin producing strains (virulent strains). Virulent strain had a significantly closer relationship with recurrent abdominal pain, while no difference was found in other symptoms. Age of patients showed a highly significant positive correlation with virulent H. pylori strain. Histopathological finding revealed that oesophagitis was significantly common in patients with negative H. pylori. Endoscopy and histopathological findings revealed that virulent strain had a significantly higher gastritis and oesophagitis rates. Conclusion Infection by virulent strains of H. pylori in children is not uncommon: It is associated with endoscopic and histopathologic changes. All cases infected with virulent strain showed positive results by using rapid urease test, culture and IgG antibodies.

66-69

Authors: KT Wariso, IM Siminialayi, JO Odigie

Objective To explore the prevalence, pathogenicity, and antibiotic susceptibility patterns of urinary tract infections at the University of Port Harcourt Teaching Hospital. Methods Samples from 400 patients with a presumptive diagnosis of urinary tract infection including 250 non pregnant females and 150 males were used for this study. They were distributed into two groups: children aged 2 to 17 (Group A) and adults aged 18 to 75 (Group B). The standard wire loop and agar diffusion technique were employed for culture and susceptibility testing, respectively. Data obtained were analysed using SPSS, version 14. Results 30.0% of Group A and 41.0% of Group B had significant bacteriuria with 66.7% and 79.3% as females, respectively. The commonest isolates cultured were Escherichia coli (32.8%), Staphylococcus aureus (17.2%), and Klebsiella spp. (16.4%). About 76.6% of isolates were sensitive to the fluorinated quinolones, 31.2% to the aminoglycosides, and 22.7% to the urinary antiseptic, nitrofurantoin. The isolates were non-sensitive to tetracycline (93.8%), cotrimoxazole (92.2%), and nalidixic acid (86.7%). Most isolates showed non-uniform sensitivity patterns to the cephaloporins (cefuroxime and ceftazidime). Pseudomonas spp. isolates were generally resistant to the fluorinated quinolones. Conclusion Though the fluorinated quinolones are still largely effective for empirical therapy in urinary tract infections, the importance of prior sensitivity testing in checking the emergence of bacterial antibiotic resistance can not be overemphasized.

74-77

Authors: Peter Felkai

The Asia–Pacific region is becoming an increasingly popular destination for holidaymakers and business travelers. Despite the seductive tourist attractions and the growing financial importance of the region, infectious diseases have unfortunately repeatedly caused major epidemics in the region. The area includes various geographical situations from high mountains to the subtropical and monsoon forests, dry deserts and seashore. The travel medicine specialist's task is to decrease the various risk of travel to the region. Infectious diseases, extreme temperatures and marine hazards could be almost entirely prevented by primary prevention forms of travel medicine such as vaccination, chemoprophylaxis and pretravel advice. There are some high–risk groups (e.g., elderly people, backpackers, families with children, adventure travelers) among the growing number of tourists. Obviously, the increased numbers of such travelers result in a rising incidence of travel–related illnesses and accidents. For those groups, primary prevention sometimes is not enough. The pretravel medical check up, specialists' consultations and treatment, as forms of secondary prevention, are also essential in travel medicine. Patients who suffer from serious illnesses or who have accidents must often be repatriated. “State–of–the–art repatriation”, as a special form of tertiary prevention of travel medicine will play an important role in the future and is directly related to effective travel insurance, which could help in providing good medical care to the patients even far from their home. The levels, methods and importance of prevention in travel medicine are discussed in this article.

81-82

Authors: Asad U Khan

Recently WHO has again declared Influenza pandemic due to the outbreak of H1N1 which has infected over 254,206 people in 80 countries, with 625 deaths. Our lab has started working on the proteome analysis of H1N1, 2009 out break strains.