Asian Pacific Journal of Tropical Biomedicine

Issue 4,2013 Table of Contents

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  • 1  Bioavailability enhancers of herbal origin: An overview
    Kritika Kesarwani Rajiv Gupta
    [Abstract](29) [HTML](0) [PDF 882.12 K](76)
    Recently, the use of herbal medicines has been increased all over the world due to their therapeutic effects and fewer adverse effects as compared to the modern medicines. However, many herbal drugs and herbal extracts despite of their impressive in-vitro findings demonstrates less or negligible in-vivo activity due to their poor lipid solubility or improper molecular size, resulting in poor absorption and hence poor bioavailability. Nowadays with the advancement in the technology, novel drug delivery systems open the door towards the development of enhancing bioavailability of herbal drug delivery systems. For last one decade many novel carriers such as liposomes, microspheres, nanoparticles, transferosomes, ethosomes, lipid based systems etc. have been reported for successful modified delivery of various herbal drugs. Many herbal compounds including quercetin, genistein, naringin, sinomenine, piperine, glycyrrhizin and nitrile glycoside have demonstrated capability to enhance the bioavailability. The objective of this review is to summarize various available novel drug delivery technologies which have been developed for delivery of drugs (herbal), and to achieve better therapeutic response. An attempt has also been made to compile a profile on bioavailability enhancers of herbal origin with the mechanism of action (wherever reported) and studies on improvement in drug bioavailability, exhibited particularly by natural compounds.
    2  Experimental evaluation of Candonocypris novaezelandiae (Crustacea: Ostracoda) in the biocontrol of Schistosomiasis mansoni transmission
    Fouad Yousif Sherif Hafez Samia El Bardicy Menerva Tadros Hoda Abu Taleb
    [Abstract](17) [HTML](0) [PDF 907.30 K](84)
    Objective: To test Candonocypris novaezelandiae (Baird) (C. novaezelandiae), sub-class Ostracoda, obtained from the Nile, Egypt for its predatory activity on snail, Biomphalaria alexandrina (B. alexandrina), intermediate host of Schistosoma mansoni (S. mansoni) and on the free-living larval stages of this parasite (miracidia and cercariae). Methods: The predatory activity of C. novaezelandiae was determined on B. alexandrina snail (several densities of eggs, newly hatched and juveniles). This activity was also determined on S. mansoni miracidia and cercariae using different volumes of water and different numbers of larvae. C. novaezelandiae was also tested for its effect on infection of snails and on the cercarial production. Results: C. novaezelandiae was found to feed on the eggs, newly hatched and juvenile snails, but with significant reduction in the consumption in the presence of other diet like the blue green algae (Nostoc muscorum). This ostracod also showed considerable predatory activity on the free-living larval stages of S. mansoni which was affected by certain environmental factors such as volume of water, density of C. novaezelandiae and number of larvae of the parasite. Conclusions: The presence of this ostracod in the aquatic habitat led to significant reduction of snail population, infection rate of snails with schistosme miracidia as well as of cercarial production from the infected snails. This may suggest that introducing C. novaezelandiae into the habitat at schistosome risky sites could suppress the transmission of the disease.
    3  Antioxidant, antibacterial and cytotoxic effects of the phytochemicals of whole Leucas aspera extract
    Md Atiar Rahman Md Saiful Islam
    [Abstract](21) [HTML](0) [PDF 874.98 K](80)
    Objective: To investigate the antioxidant, antibacterial and cytotoxic activity of whole Leucas aspera (Labiatae) (L. aspera) alcoholic extract. Methods: Whole L. aspera powder was extracted by absolute ethanol (99.50%). The ethanolic extract was subjected to antioxidant, antibacterial and brine shrimp lethality assay. Results: The extract showed potent radical scavenging effect (antioxidant) with IC50 value of (99.58±1.22) µg/mL which was significant (P<0.01) in comparison to ascorbic acid with IC50 value of (1.25±0.95) µg/mL. In case of antibacterial screening, the extract showed notable antibacterial effect against the tested microbial strains. Significant (P<0.05) zone of inhibitions against Gram positive Bacillus subtilis [(12.00±1.32) mm] and Bacillus megaterium [(13.00±1.50) mm], Staphylococcus aureus [(8.00±0.50) mm] and Gram negative Salmonella typhi [(6.00±0.50) mm], Salmonella paratyphi [(8.00±1.00) mm], Shigella dysenteriae [(9.00±1.32) mm] and Vibrio cholerae [(9.00±0.66) mm] was observed. In brine shrimp lethality bioassay, the extract showed the LC50 value as (181.6±2.15) µg/mL which was statistically significant (P<0.01) compared to positive control vincristine sulfate [LC50=(0.76±0.04) µg/mL]. Conclusions: The results demonstrate that the ethanolic extract of L. aspera could be used as antibacterial, pesticidal and various pharmacologic actives.
    4  Antipyretic and anticonvulsant activity of n-hexane fraction of Viola betonicifolia
    Naveed Muhammad Muhammad Saeed Haroon Khan Naila Raziq Syed Muhammad Ashhad Halimi
    [Abstract](18) [HTML](0) [PDF 885.60 K](77)
    Objective: To investigate the antipyretic and anticonvulsant activities of n-hexane fraction of Viola betonicifolia (V. betonicifolia). Methods: The antipyretic effect was scrutinized using brewer’s yeast induced pyrexia and anticonvlsion effect was tested using pentylenetetrazol and strychnine induced convulsion in mice. Results: N-hexane fraction of V. betonicifolia demonstrated highly significant antipyretic activity during various assessment times (1-5 h) when challenged in yeast induced pyrexia test. The effect was in a dose dependent manner with maximum attenuation (82.50%) observed at 300 mg/kg i.p. When tested in pentylenetetrazol induced convulsion test, the 1st stage (Ear and facial twitching) and 2nd stage (Convulsive wave through the body) was 100% protected during 24 h at all the test doses (300, 400 and 500 mg/kg i.p.), while the latency time of remaining stages was significantly increased. The maximum effect was observed by n-hexane fraction of V. betonicifolia at 400 and 500 mg/kg i.p., as the latency time for generalized clonic-tonic seizure (5th stage) was increased up to 25.34 min. However, n-hexane fraction of V. betonicifolia had no protection in strychnine induced convulsion test. Conclusions: In conclusion, phytopharmacological studies provide scientific foundation to the folk uses of the plant in the treatment of pyrexia and neurological disorders.
    5  The identities and anti-herpes simplex virus activity of Clinacanthus nutans and Clinacanthus siamensis
    Paween Kunsorn Nijsiri Ruangrungsi Vimolmas Lipipun Ariya Khanboon Kanchana Rungsihirunrat
    [Abstract](9) [HTML](0) [PDF 1.71 M](82)
    Objective: To distinguish the difference among the Clinacanthus nutans (Burm. f.) Lindau (C. nutans) and Clinacanthus siamensis Bremek (C. siamensis) by assessing pharmacognosy characteristics, molecular aspect and also to evaluate their anti-herpes simplex virus (HSV) type 1 and type 2 activities. Methods: Macroscopic and microscopic evaluation were performed according to WHO Geneva guideline. Stomatal number, stomatal index and palisade ratio of leaves were evaluated. Genomic DNA was extracted by modified CTAB method and ITS region was amplified using PCR and then sequenced. Dry leaves were subsequently extracted with n-hexane, dichloromethane and methanol and antiviral activity was performed using plaque reduction assay and the cytotoxicity of the extracts on Vero cells was determined by MTT assay. Results: Cross section of midrib and stem showed similar major components. Leaf measurement index of stomatal number, stomatal index and palisade ratio of C. nutans were 168.32±29.49, 13.83±0.86 and 6.84±0.66, respectively, while C. siamensis were 161.60±18.04, 11.93±0.81 and 3.37±0.31, respectively. The PCR amplification of ITS region generated the PCR product approximately 700 bp in size. There were 34 polymorphisms within the ITS region which consisted of 11 Indels and 23 nucleotide substitutions. The IC50 values of C. nutans extracted with n-hexane, dichloromethane and methanol against HSV-1 were (32.05±3.63) µg/mL, (44.50±2.66) µg/mL, (64.93± 7.00) µg/mL, respectively where asthose of C. siamensis were (60.00±11.61) µg/mL, (55.69±4.41) µg/mL, (37.39 ±5.85) µg/mL, respectively. Anti HSV-2 activity of n-hexane, dichloromethane and methanol C. nutans leaves extracts were (72.62±12.60) µg/mL, (65.19±21.45) µg/mL, (65.13±2.22) µg/mL, respectively where as those of C. siamensis were (46.52±4.08) µg/mL, (49.63±2.59) µg/mL, (72.64±6.52) µg/mL, respectively. Conclusions: The combination of macroscopic, microscopic and biomolecular method are able to authenticate these closely related plants and both of them have a potency to be an anti-HSV agent.
    6  Cytotoxic and antibacterial substances against multi-drug resistant pathogens from marine sponge symbiont: Citrinin, a secondary metabolite of Penicillium sp.
    Ramesh Subramani Rohitesh Kumar Pritesh Prasad William Aalbersberg
    [Abstract](19) [HTML](0) [PDF 1.02 M](77)
    Objective: To Isolate, purify, characterize, and evaluate the bioactive compounds from the sponge-derived fungus Penicillium sp. FF001 and to elucidate its structure. Methods: The fungal strain FF001 with an interesting bioactivity profile was isolated from a marine Fijian sponge Melophlus sp. Based on conidiophores aggregation, conidia development and mycelia morphological characteristics, the isolate FF001 was classically identified as a Penicillium sp. The bioactive compound was identified using various spectral analysis of UV, high resolution electrospray ionization mass spectra, 1H and 13C NMR spectral data. Further minimum inhibitory concentrations (MICs) assay and brine shrimp cytotoxicity assay were also carried out to evaluate the biological properties of the purified compound. Results: Bioassay guided fractionation of the EtOAc extract of a static culture of this Penicillium sp. by different chromatographic methods led the isolation of an antibacterial, anticryptococcal and cytotoxic active compound, which was identified as citrinin (1). Further, citrinin (1) is reported for its potent antibacterial activity against methicillin-resistant Staphylococcus aureus (S. aureus), rifampicin-resistant S. aureus, wild type S. aureus and vancomycin-resistant Enterococcus faecium showed MICs of 3.90, 0.97, 1.95 and 7.81 µg/mL, respectively. Further citrinin (1) displayed significant activity against the pathogenic yeast Cryptococcus neoformans (MIC 3.90 µg/mL), and exhibited cytotoxicity against brine shrimp larvae LD50 of 96 µg/mL. Conclusions: Citrinin (1) is reported from sponge associated Penicillium sp. from this study and for its strong antibacterial activity against multi-drug resistant human pathogens including cytotoxicity against brine shrimp larvae, which indicated that sponge associated Penicillium spp. are promising sources of natural bioactive metabolites.
    7  Combinational effects of hexane insoluble fraction of Ficus septica Burm. F. and doxorubicin chemotherapy on T47D breast cancer cells
    Agung Endro Nugroho Adam Hermawan Dyaningtyas Dewi Pamungkas Putri Anindya Novika Edy Meiyanto
    [Abstract](20) [HTML](0) [PDF 1.44 M](78)
    Objective: To evaluate the effects of n-hexane insoluble fraction (HIF) of Ficus septica leaves in combination with doxorubicin on cytotoxicity, cell cycle and apoptosis induction of breast cancer T47D cell lines. Methods: The in vitro drugs-stimulated cytotoxic effects were determined using MTT assay. Analysis of cell cycle distribution was performed using flowcytometer and the data was analyzed using ModFit LT 3.0 program. Apoptosis assay was carried out by double staining method using ethydium bromide-acridin orange. The expression of cleaved-poly (ADP-ribose) polymerase (PARP) on T47D cell lines was identified using immunocytochemistry. Results: The combination exhibited higher inhibitory effect on cell growth than the single treatment of doxorubicin in T47D cells. In addition, combination of doxorubicin and HIF increased the incidence of cells undergoing apoptosis. HIF could improve doxorubicin cytotoxic effect by changing the accumulation of cell cycle phase from G2/M to G1 phase. The combination also exhibited upregulation of cleaved-PARP in T47D cells. Conclusions: Based on this results, HIF is potential to be developed as co-chemotherapeutic agent for breast cancer by inducing apoptosis and cell cycle arrest. However, the molecular mechanism need to be explored further.
    8  An abattoir survey of contagious bovine pleuropneumonia lesions in slaughtered cattle in selected districts in Northern Tanzania
    Emmanuel Swai Isidory Mwezimpya Edward Ulicky Adam Mbise Winford Moshy
    [Abstract](33) [HTML](0) [PDF 824.85 K](78)
    Objective: To establish and estimate incidence of contagious bovine pleuropneumonia (CBPP), using abattoir survey as a diagnostic tool in slaughtered cattle in Northern Tanzania. Methods: A total of 4 460 cattle were slaughtered in five abattoirs in 3 northern zone regions (Arusha, Kilimanjaro and Tanga) during the period of January to May 2004. They were examined ante- mortem for ‘pneumonia signs’, and ‘characteristic contagious bovine pleuropneumonia (CBPP) lung lesions’. Results: Forty-one (0.91%) of the slaughtered cattle, the majority of which were Tanzania short horn zebu, had gross lung lesions suggestive of CBPP. The prevalence of lesions was significantly (P<0.05) higher in Karatu abattoir compared to others. No animal was detected to have lesion in Bomang’ ombe abattoir. The most observed pneumonic signs included labored breathing (90%), dry cough (57%) and mucopurulent nasal discharge (47%). The gross characteristic CBPP pathological lesion, frequently encountered was left lung lesion (47%), pinkish lung (71%) and pleural adhesion (98%). Epidemiological reports show that the CBPP reported outbreaks increased from 19 in 2002, 65 in 2003 and 18 in 2004 (January-March). The corresponding number of reported deaths increased from 137 in 2002, 269 in 2003 and 77 in 2004 (January-March). Conclusions: It’s concluded from this study that CBPP is a problem in spite of the extensive awareness and vaccination campaigns. Nevertheless, a continued surveillance programme including routine checks of all cattle carcasses at the abattoir and subsequent epidemiological investigation of suspected cases are recommended.
    9  Urinary schistosomiasis and malaria associated anemia in Ethiopia
    Ketema Deribew Zinaye Tekeste Beyene Petros
    [Abstract](24) [HTML](0) [PDF 782.29 K](74)
    Objective: To assess the prevalence of anemia in children with urinary schistosomiasis, malaria and concurrent infections by the two diseases. Methods: Urine and blood samples were collected from 387 children (216 males and 171 females) to examine urinary schistosomiasis and malaria and to determine hemoglobin concentration at Hassoba and Hassoba Buri village in Amibara woreda, Afar region, Ethiopia. Results: The overall prevalence of urinary schistosomiasis and Plasmodium falciparum malaria was 24.54% and 6.20% respectively. Only 2.84% of children carried concurrent infections of both parasites. There was high percentage of anemic patients (81.81%) in the coinfected cases than in either malaria (33.3%) or schistosomiasis (38.94%) cases. There was significantly low mean hemoglobin concentration in concurrently infected children than non-infected and single infected (P<0.05). The mean hemoglobin concentration between Plasmodium falciparum and S. haematobium infected children showed no significant difference (P>0.05). The level of hemoglobin was negatively correlated with the number of S. haematobium eggs/10 mL urine (r=-0.6) and malaria parasitemia (r=-0.53). Conclusions: The study showed that anemia is higher in concurrently infected children than non-infected and single infected. Furthermore, level of hemoglobin was negatively correlated with the number of S. haematobium eggs and malaria parsitemia. Therefore, examination of hemoglobin status in patients co-infected with malaria and schistosomiasis is important to reduce the risk of anemia and to improve health of the community.
    10  Adult female of Strongyloides stercoralis in respiratory secretions
    Bava Amadeo Javier Bava Domínguez Cecilia Troncoso Alcides
    [Abstract](9) [HTML](0) [PDF 300.87 K](76)
    Objective: To communicate the presence of adult females, rabditoid larvae and eggs of Strongyloides stercoralis (S. stercoralis) in the respiratory secretions obtained by tracheal aspirate from a HIV-negative patient who was suffering from polymyositis, and treated with corticoids and amethopterin and assisted by pneumonia. Methods: The respiratory secretions submitted to the Parasitology Laboratory of the Muñiz Hospital were made more concentrated by centrifugation (1 500 r/min for 15 seconds). Wet mount microscopy was performed with the pellet. Results: It revealed adult females, rabditoid larvae and eggs of S. stercoralis. Further parasitological studies performed after the start of the treatment with ivermectin on fresh fecal samples, gastric lavages and tracheal aspirates showed scanty mobile filariform and rabditoid larvae of the same parasite. Conclusions: The presence of adult female S. stercoralis which has never been observed before in the clinical samples submitted to our Laboratory for investigation can be considered as an indirect marker of the severe immunosupression of the patient.
    11  Discovery of Trichuris landak n. sp. by Endang Purwaningsih
    Lim Boon Huat
    [Abstract](6) [HTML](0) [PDF 610.76 K](73)
    12  Surveillance of multidrug resistant uropathogenic bacteria in hospitalized patientsinIndian
    Monali Priyadarsini Mishra Nagen Kumar Debata Rabindra Nath Padhy
    2013(4):315-324. DOI: 10.1016/S2221-1691(13)60071-4
    [Abstract](16) [HTML](0) [PDF 892.43 K](73)
    Objective: To record surveillance, antibiotic resistance of uropathogens of hospitalized patients over a period of 18 months. Methods: Urine samples from wards and cabins were used for isolating urinary tract infection (UTI)-causing bacteria that were cultured on suitable selective media and identified by biochemical tests; and their antibiograms were ascertained by Kirby- Bauer’s disc diffusion method, in each 6-month interval of the study period, using 18 antibiotics of five different classes. Results: From wards and cabins, 1 245 samples were collected, from which 996 strains of bacteria belonging to 11 species were isolated, during April 2011 to September 2012. Two Gram-positive, Staphylococcus aureus (S. aureus) and Enterococcus faecalis (E. faecalis), and nine Gram-negative bacteria, Acinetobacter baumannii, Citrobacter sp., Escherichia coli, Enterobacter aerogenes, Klebsiella pneumoniae, Klebsiella oxytoca, Proteus mirabilis, Proteus vulgaris and Pseudomonas aeruginosa were isolated. Both S. aureus and E. faecalis were vancomycin resistant, and resistant-strains of all pathogens increased in each 6-month period of study. Particularly, all Gram-negatives were resistant to nitrofurantoin and co-trimoxazole, the most preferred antibiotics of empiric therapy for UTI. Conclusions: Antibiograms of 11 UTI- causing bacteria recorded in this study indicated moderately higher numbers of strains resistant to each antibiotic studied, generating the fear of precipitating fervent episodes in public health particularly with bacteria, Acinetobacter baumannii, Escherichia coli, Klebsiella pneumoniae and S. aureus. Moreover, vancomycin resistance in strains of S. aureus and E. faecalis is a matter of concern.
    13  A report of left dorsal displacement of the large colon in a tropical horse
    Farhang Sasani Javad Javanbakht Mehdi Ghamsari Mehdi Aghamohammad Hassan
    [Abstract](20) [HTML](0) [PDF 912.34 K](70)
    We report one such case which was diagnosed intraoperatively as left dorsal colon volvulus due to multiple mesenteric abnormalities. A 17-year-old castrated male horse was taken to the Tehran University Veterinary Hospital for treatment of metacarpal wound accompanied by severe abdominal distension and acute colic. The treatment and measurement were taken for a month, and the prepared biopsy indicated that the healing trend was obvious. Unfortunately, prior to discharge, the clinical colic manifestations emerged and the animal suddenly died. Dilated large intestine was palpated per rectum and a ventral midline exploratory laparotomy was performed, a complete volvulus of the ascending colon was identified with multiple mesenteric anomalies of unknown aetiology. The pathologic changes observed in this study accurately reflect those changes reported in horse with naturally occurring colonic volvulus and can serve as a reference for subsequent studies on attenuating bowel injury. The present study results can be used to make a scientific assessment of prognosis in the pre-operative, operative, and post-operative management of horses with large colon volvulus.
    14  Utilization of multiple "omics" studies in microbial pathogeny for microbiology insights
    Viroj Wiwanitkit
    [Abstract](20) [HTML](0) [PDF 661.16 K](78)
    In the present day, bioinformatics becomes the modern science with several advantages. Several new “omics” sciences have been introduced for a few years and those sciences can be applied in biomedical work. Here, the author will summarize and discuss on important applications of omics studies in microbiology focusing on microbial pathogeny. It can be seen that genomics and proteinomics can be well used in this area of biomedical studies.
    15  Prevalence survey of infection with Treponema pallidum among HIV-positive patients in Tehran
    Banafsheh Moradmand Badie Zeinab Yavari Shooka Esmaeeli Koosha Paydary Sahra Emamzadeh-Fard SeyedAhmad SeyedAlinaghi Mehrnaz Rasoulinejad
    [Abstract](20) [HTML](0) [PDF 648.28 K](78)
    Objective: To identify the frequency of syphilis among Iranian HIV-positive patients. Methods: A cross-sectional study on the prevalence of syphilis and HIV co-infection among 450 patients diagnosed with HIV infection was conducted between 2004 and 2008 at Imam Khomeini hospital, Tehran, Iran. The lab tests including CD4 cell count, cerebrospinal fluid, veneral disease research laboratory (VDRL), fluorescent treponema antibody-absorption (FTA-Abs) and viral load were performed for all the patients. Data regarding medical history and their demographics were also collected. Results: Of all 450 HIV-positive patients, 24 (5.3%) had a positive VDRL test and only two men had a FTA-Abs positive test which means 0.45% of them had a definite co-infection of syphilis. 65.3% of the HIV-positive patients were injection drug users that the co-infection prevalence of them was 0.7%. We did not find any patient with neurosyphilis. Conclusions: Considering the increasing prevalence of HIV and also extensive use of highly active antiretroviral therapy in developing nations, the diagnosis of syphilis should be timely established using screening tests among such patients.

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