Synthesis, DNA binding and antimicrobial studies on rhodium(II) complexes of dicyandiamide.

The synthesis and characterization on four rhodium(II) complexes with the formula [Rh2(CH3COO)2(AMUH)2(dcda)2](CH3COO)2(1),[Rh2(CH3COO)2(AEUH)2(dcda)2](CH3COO)2(2),[Rh2(CH3COO)2(APrnUH)2(dcda)2](CH3COO)2(3),[Rh2(CH3COO)2(ABnUH)2(dcda)2](CH3COO)2(4), where AMUH = 1-amidino-O-methylurea, AEUH = 1-amidino-O-ethylurea, APrnUH = 1-amidino-O-n-propylurea, ABnUH = 1-amidino-O-n-butylurea, dcda = dicyandiamide are reported. The complexes were prepared by the reaction of dicyandiamide with rhodium(II) acetate in methanol (1), ethanol (2), n-propanol (3) and n-butanol (4) respectively and characterized by various techniques such as C, H, N analysis, FTIR, UV-Visible, EPR, conductance, SEM, EDX, powder XRD pattern and mass spectral studies. The interaction studies of the complexes with CT-DNA suggested the non-intercalative mode of binding for these complexes. The antimicrobial activity of the complexes against the tested microorganisms viz. Bacillus subtilis, Enterococcus faecium, Staphylococcus aureus and Escherichia coli, using the standard antibiotics streptomycin as positive control is also reported.

Dengue in western Terai region of Nepal.

BACKGROUND: Dengue Fever (DF) is an emerging mosquito-borne disease. It is a nagging public health problem in the low lands of Terai, expanding to new areas of Nepal in recent years. METHODS: A cross-sectional study was conducted to determine anti-Dengue IgM positive rate in Mahendranagar, Dhangadi and Dang between August 2008 and November 2009. Serum samples were collected from 283 patients visiting hospitals with history of fever, headache and suspected DF. The samples were examined by ELISA. RESULTS: The anti-Dengue IgM positivity was found to be 9.8 %. The positive rate was highest in Mahendranagar (13.3 %) followed by Dhangadi (9.8 %) (P<0.05). The Dengue positive cases were higher in female (10.9 %) than males (9.0 %). The positivity was higher in Ethnic group Brahman/Chherti (13.1%) as compared to Janajati (5.6 %). The highest positive cases (10.7 %) were from age group above 50 years. The highest numbers of Dengue positive cases were observed in occupation group of agriculture (18.2 %) (P>0.05). CONCLUSIONS: Dengue has substantial expansion in Western and Far Western Terai region of Nepal which was limited to the middle Terai region in the past and mostly infects older people.

Knowledge and practices of pig farmers regarding Japanese encephalitis in Kathmandu, Nepal.

Japanese encephalitis (JE) is the single largest cause of viral encephalitis in the world and has been endemic in Nepal since the early 1980s. Since then, it has spread from its origins in lowland plains to the Kathmandu Valley as well as in hill and mountain districts. Pigs are amplifying hosts for the virus. The Nepal government has been encouraging the development of pig farming as a means of poverty alleviation. Whereas other countries have reduced JE through vaccination programmes and improvements in pig husbandry, these options are not economically possible in Nepal. The objective of this study was to examine the occupational risk of pig farmers in Nepal and to determine their level of knowledge and practice of JE prevention techniques. We surveyed 100 randomly selected pig farmers in the Kathmandu District and found that pig farmers were exposed to many JE risk factors including poverty and close proximity to pigs, rice paddy fields and water birds, which are the definitive hosts for the virus. Forty-two percent of the farmers had heard of JE, 20% associated it with mosquito bites and 7% named pigs as risk factors. Few protective measures were taken. None of the farmers were vaccinated against JE nor were any pigs, despite an ongoing human vaccination campaign. This farming community had little ownership of land and limited education. JE education programmes must consider gender differences in access to public health information as there were an equal number of male and female farmers. We provide findings that can inform future JE education programmes for this vulnerable population.

An evaluation of patients care in Radio Diagnosis Department of Tribhuvan University Teaching Hospital, Kathmandu, Nepal.

Patients care involves all the activities that are carried out before, during and after radiological diagnostic procedures. The successful application of medical knowledge depends on what patients think and feel about the medical personnel and the hospital. From the clinical experience, patients usually reacts to some factors that creates problems in the radiology units such as delay, neglect, use of harsh words, unnecessary repeats and preferential treatment. The study was aimed at evaluating patients care in radio diagnosis department of Tribhuvan University Teaching Hospital (TUTH), Kathmandu, Nepal. A cross sectional study was conducted from May to June, 2012 in TUTH, Kathmandu, Nepal. Semi structured both open and closed ended questionnaires were administered among 287 conscious patients who were agreed to participate in the study. A total 287 patients were enrolled in the study. Out of them, 165 (57.49%) patients perceived that instruction was adequate before and after the examination; 219 (76.30%) patients were satisfied with the services rendered in the hospital; 174 (60.62%) patients perceived that privacy was adequate but 229 (79.9%) and 145 (50.5%) perceived that waiting areas and social aspects of patients care were suboptimal and in adequate. This can be improved by hospital management system.

Serological evidence of Trichinellosis in local pigs of Nepal.

In Nepal, animal husbandry is a major source of income. Pig husbandry is practiced in rural, peri-urban, and urban communities. Free ranging "back yard" pigs and the practice of feeding offal is a very common management practice which potentially allows for the transmission of trichinellosis; however, this zoonosis has never been reported from this region. A total of 425 serum samples were collected from local pigs. These were initially screened by ELISA after which positive samples were examined by Western blot. This procedure identified two samples which had clear specific bands for Trichinella; however, muscle samples tested by HCL-pepsin digestion were found to be negative. If these highly specific serological analyses are confirmed, this would be the first report of trichinellosis in Nepal and a prevention program should be initiated to limit the access of pigs to open garbage dumps which exist both in towns and on farms.

Prevalence of pulmonary tuberculosis in far Western Nepal.

Keeping in view of high prevalence of tuberculosis in far western development region in different reports, a door to door survey of 125 household was carried out randomly in Mahendra Nagar municipality ward no 13 of Kanchanpur district to pick up symptomatics suggestive of pulmonary tuberculosis and find the prevalence in the target population. A total of 200 sputum specimens from 70 symptomatics within the age range of 10-80 years were collected and subjected to smear microscopy by Ziehl - Neelsen's method. Out of 10 positive cases (14.28%), infection in males and females was 80% and 20% respectively. The infection was found in the productive age group 20-69. The highest prevalence was obtained in 20-29 age group (25%) followed by 40-49 (11%), 30-39 (9%), 50-59 (8%) and 60-69 (8%). The diagnostic yield of single sputum specimen examined was insufficient in field condition, especially where the sputum positivity was low. Of the three sputum smears examined, the early morning specimen had the best result.

Negative feedback on the effects of stem cell factor on hematopoiesis is partly mediated through neutral endopeptidase activity on substance P: a combined functional and proteomic study.

Hematopoietic regulation is a complex but dynamic process regulated by intercellular and intracellular interactions within the bone marrow (BM) microenvironment. Through neurokinin-1 (NK-1) and NK-2 receptors, peptides (eg, substance P [SP]) encoded by the preprotachykinin-I gene mediate distinct hematopoietic effects. Cytokines, associated with hematopoietic stimulation, and SP regulate the expression of each other in BM mesenchymal and immune cells. Neutral endopeptidase (NEP) uses SP as a substrate to produce SP(1-4), which inhibits the proliferation of matured myeloid progenitor. This study determines whether the degradation of SP to SP(1-4) by endogenous NEP in BM stroma could be a feedback on hematopoietic stimulation by stem cell factor (SCF). SP(1-4) induced the production of transforming growth factor (TGF)-beta and tumor necrosis factor-alpha in BM stroma. TGF-beta production accounted for part of the inhibitory effects by SP(1-4) on the proliferation of early (granulocyte-macrophage colony-forming units) and late (long-term culture-initiating cells) hematopoietic progenitors. Enzyme-linked immunosorbent assays and/or protein-chip arrays indicated a timeline change of SP to SP(1-4) in BM stroma stimulated with SCF, which correlated with increase in NEP messenger RNA. Since SP and its fragment, SP(1-4), interact with the same receptor to mediate opposing hematopoietic effects, 2 interactive studies were done to understand the dual responses of NK-1: (1) a 3-dimensional molecular model of NK-1 and SP and (2) screening of a random dodecapeptide library for SP(1-4) interacting sites. The effects of SP(1-4) on hematopoietic progenitors and the timeline change of SP to SP(1-4), together with the 3-dimensional model, provide a partial explanation for the feedback on the stimulatory effects of SCF and SP on hematopoiesis.

Mimicry between neurokinin-1 and fibronectin may explain the transport and stability of increased substance P immunoreactivity in patients with bone marrow fibrosis.

Bone marrow (BM) fibrosis may occur in myeloproliferative diseases, lymphoma, myelodysplastic syndrome, myeloma, and infectious diseases. In this study, the role of substance P (SP), a peptide with pleiotropic functions, was examined. Some of its functions-angiogenesis, fibroblast proliferation, and stimulation of BM progenitors-are amenable to inducing BM fibrosis. Indeed, a significant increase was found in SP-immunoreactivity (SP-IR) in the sera of patients with BM fibrosis (n = 44) compared with the sera of patients with hematologic disorders and no histologic evidence of fibrosis (n = 46) (140 +/-12 vs 18 +/-3; P <.01). Immunoprecipitation of sera SP indicated that this peptide exists in the form of a complex with other molecule(s). It was, therefore, hypothesized that SP might be complexed with NK-1, its natural receptor, or with a molecule homologous to NK-1. To address this, 3 cDNA libraries were screened that were constructed from pooled BM stroma or mononuclear cells with an NK-1 cDNA probe. A partial clone (clone 1) was retrieved that was 97% homologous to the ED-A region of fibronectin (FN). Furthermore, sequence analyses indicated that clone 1 shared significant homology with exon 5 of NK-1. Immunoprecipitation and Western blot analysis indicated co-migration of SP and FN in 27 of 31 patients with BM fibrosis. Computer-assisted molecular modeling suggested that similar secondary structural features between FN and NK-1 and the relative electrostatic charge might explain a complex formed between FN (negative) and SP (positive). This study suggests that SP may be implicated in the pathophysiology of myelofibrosis, though its role would have to be substantiated in future research. (Blood. 2001;97:3025-3031)

Induction of preprotachykinin-I and neurokinin-1 by adrenocorticotropin and prolactin. Implication for neuroendocrine-immune-hematopoietic axis.

We studied the complex interactions within the neuroendocrine-immune-hematopoietic axis by determining a possible link among ACTH, PRL, PPT-I and the receptors for its peptides, NK-1 and NK-2. Indeed, ACTH and PRL induced the expression of PPT-I and NK-1 in human bone marrow stroma with no effect on NK-2. Consistent with a role for PPT-I in regulating the development of myeloid and erythroid progenitors, we found that ACTH and PRL, through NK-1 stimulated the proliferation of both types of progenitors. Induction of PPT-I was regulated at the transcriptional and post-transcriptional levels. The results showed that ACTH and PRL stimulated the proliferation of bone marrow progenitors, partly through PPT-I and NK-1 induction.

Health promotion approach for the control of food-borne parasitic zoonoses in Nepal: emphasis on an environmental assessment.

Green and Kreuter define health promotion as the use of both education and ecology to encourage and support living conditions conducive to good health. Their PRECEDE-PROCEED model delineates a practical way to cope with health problems using this definition. Applying PRECEDE to Nepal helps identify the steps needed to control the ever-increasing incidence of food-borne parasitic zoonoses (FBPZ) there and in other South Asian countries. In executing the first five steps of the model, we focused on behavioral and environmental assessments to find a method for controlling FBPZ. Through these two assessments, we identified the following behavioral and environmental objectives: establish a meat inspection system by 2003, establish training programs on safe meat production and selling practices by 2003, improve slaughterhouses and slaughtering practices by 2003. The educational and ecological assessments revealed that the necessary predisposing, reinforcing and enabling factors for appropriate control of FBPZ are present in Nepal, while an administration and policy assessment shows Nepal meets PRECEDE requirements through its recent meat inspection legislation. Although the data for each element of the PRECEDE-PROCEED model are limited in Nepal, they clearly tell us to move forward to the PROCEED stage to control FBPZ there as well as in all of South Asia.

Controlling Taenia solium in Nepal using the PRECEDE-PROCEED model.

Taenia solium is a predominant food-borne parasitic zoonosis (FBPZ) in Nepal. Using the PRECEDE framework, as defined by Green and Kreuter, we can identify the factors behind the high incidence of this disease. Armed with this information, we can define the actions necessary to control T. solium. In accordance with the first step of PRECEDE, social assessment, we set the goal of decreasing the potential for T. solium transmission in Nepal by the year 2003. This goal has yet to be endorsed by policy makers. However, an epidemiological assessment based on hospital data alone showed that T. solium is an endemic problem in urban Nepal that must be addressed. Based on behavioral and environmental assessments (Steps 1 and 2 of PRECEDE-PROCEED), we defined the following action objectives to be achieved by 2003: 1) Train meat producers and sellers to detect contaminated pork and avoid selling it, 2) Improve pig husbandry to limit the animals' access to human feces, 3) Construct hygienic model slaughterhouses. These improvements could control the meat-producing environment, thus limiting the potential for cross-carcass contamination and other hygiene deficiencies associated with the spread of T. solium. An educational and ecological assessment shows all predisposing, reinforcing and enabling factors are present in Nepal, consistent with PRECEDE requirements. While T. solium is clearly defined as a health problem according to PRECEDE, there remain significant hurdles to controlling it. These hurdles lie in administration and policy, where standardized law-enforcement and meat inspection practices are needed. Finally, the government of Nepal must assign high priority to T. solium control, as it is a preventable, yet prevalent disease.

Increased expression of preprotachykinin-I and neurokinin receptors in human breast cancer cells: implications for bone marrow metastasis.

Neuropeptides are implicated in many tumors, breast cancer (BC) included. Preprotachykinin-I (PPT-I) encodes multiple neuropeptides with pleiotropic functions such as neurotransmission, immune/hematopoietic modulation, angiogenesis, and mitogenesis. PPT-I is constitutively expressed in some tumors. In this study, we investigated a role for PPT-I and its receptors, neurokinin-1 (NK-1) and NK-2, in BC by using quantitative reverse transcription-PCR, ELISA, and in situ hybridization. Compared with normal mammary epithelial cells (n = 2) and benign breast biopsies (n = 21), BC cell lines (n = 7) and malignant breast biopsies (n = 25) showed increased expression of PPT-I and NK-1. NK-2 levels were high in normal and malignant cells. Specific NK-1 and NK-2 antagonists inhibited BC cell proliferation, suggesting autocrine and/or intercrine stimulation of BC cells by PPT-I peptides. NK-2 showed no effect on the proliferation of normal cells but mediated the proliferation of BC cells. Cytosolic extracts from malignant BC cells enhanced PPT-I translation whereas extracts from normal mammary epithelial cells caused no change. These enhancing effects may be protein-specific because a similar increase was observed for IL-6 translation and no effect was observed for IL-1alpha and stem cell factor. The data suggest that PPT-I peptides and their receptors may be important in BC development. Considering that PPT-I peptides are hematopoietic modulators, these results could be extended to understand early integration of BC cells in the bone marrow, a preferred site of metastasis. Molecular signaling transduced by PPT-I peptides and the mechanism that enhances translation of PPT-I mRNA could lead to innovative strategies for BC treatments and metastasis.

Effects of preprotachykinin-I peptides on hematopoietic homeostasis. A role for bone marrow endopeptidases.

Hematopoiesis is maintained by "fine-tuned" regulation among cytokines, neuropeptides, neurotransmitters, and neurotrophic factors. Neurotransmitters, derived from PPT-I exert immune and hematopoietic regulation. PPT-I is also expressed locally in bone marrow (BM) stromal cells. PPT-I peptides induce the production of cytokines in BM cells, resulting in regulation of both committed progenitors (CFU-GM) and primitive hematopoietic progenitors (CAFC). Both types of progenitors are regulated differently by the two major PPT-I peptides, SP and NK-A. Endopeptidases, present in BM cells, can digest SP to produce SP(1-4) and SP(4-11). In this study, we investigated the hematopoietic effects of these fragments on CFU-GM and CAFC. Similar to the two major intact PPT-I peptides (SP and NK-A), we observed different hematopoietic effects by SP(1-4) and SP(4-11). Whereas SP(1-4) exerted inhibitory effects on CFU-GM and CAFC, SP(4-11) mediated stimulatory effects. Similar to NK-A, the inhibitory effects of SP(1-4) can be partly explained by the induction of suppressive cytokines (TGF-beta, TNF-alpha, and INF-gamma). Use of antagonists and screening of a dodecapeptide expression library determined that the effects of SP(1-4) were mediated by NK-1. These results show that PPT-I peptides and their endopeptidase-derived fragments may add to the fine-tuned regulation on hematopoiesis. Furthermore, PPT-I may be exerting autoregulation to protect hematopoietic stem cells. These studies have relevance to stem cell protection and BM transplant.

Differential serodiagnosis for cystic and alveolar echinococcosis using fractions of Echinococcus granulosus cyst fluid (antigen B) and E. multilocularis protoscolex (EM18).

Echinococcus granulosus cyst fluid and E. multilocularis protoscolex extract were fractionated by a single step of preparative isoelectric focusing, resulting in an antigen B-rich fraction (8-kD) and an Em18-rich fraction, respectively. The usefulness of both fractions for differential serodiagnosis of cystic (CE) and alveolar (AE) echinococcosis was evaluated by a large-scale immunoblot analysis on a battery of 354 serum samples. These included 66 from AE patients originating from four different endemic areas, 173 from CE patients originating from seven different endemic areas, 71 from patients with other parasitic diseases, 15 from patients with hepatomas, and 29 from healthy individuals. In an immunoblot with the antigen B-rich fraction, 92% (158 of 173) of the CE sera as well as 79% (52 of 66) of the AE sera reacted with the 8-kD subunit. No cross-reactivity occurred with any sera from patients with cysticercosis, other parasitic diseases, or with hepatomas, or from healthy controls. In an immunoblot with the Em18-rich fraction, all but two sera from AE patients (64 of 66, 97%) recognized Em18, and only nine of 34 CE sera from China reacted with it. All other (139) CE sera from six other countries were negative as were all (115) other non-echinococcosis sera. These findings indicate that antigen B (8-kD) is not species-specific for E. granulosus but is genus-specific for Echinococcus, and that the Em18 antigen is a reliable serologic marker for species-specific differentiation of AE from CE.

Epidemiology of echinococcosis in Nepal.

Echinococcosis and hydatidosis is a parasitic zoonotic disease of human and animals. This disease has created public health and environment problems in all urban areas of Nepal. Based on the three year study (1993-1995) it has been revealed that the epidemiological cycle (indigenous) of Echinococcus granulosus parasite is dog-pig-dog cycle and human acquire infection accidentally through infected dog stool. However, this study has proved also the epidemiological cycle like dog-sheep-dog, dog-goat-dog and dog-buffalo-dog. This study was supported by International Development Research Centre (IDRC), Ottawa, Canada.

Echinococcus granulosus infections in the dogs of Kathmandu, Nepal.

The prevalence, incidence and risk factors of infection by Echinococcus granulosus were studied in the domestic and street dogs of Kathmandu, Nepal. Using an ELISA coproantigen test as a screening method, the highest prevalence (5/88 = 5.7%) was seen in domestic dogs from an area of the city used for slaughtering livestock. A prevalence of 1.8% (3/171) was found in domestic dogs seen at eight veterinary clinics distributed around the city. Although none of the 73 street dogs sampled in the ELISA screening survey was positive, three of 20 street dogs killed with poison as part of the city's dog-control programme harboured from one to five adult worms. As none of 99 dogs treated with an anthelmintic was found re-infected 3 months later, it was impossible to calculate accurately the of infection over a 3-month period. Information about the feeding, sleeping and roaming practices of the dogs was also gathered using questionnaires and direct observations.

Organisation of veterinary public health in the south Asia region.

In the South Asia region vast human populations are exposed daily and with considerable intensity to close contact with vast animal populations and their excreta. There is no veterinary public health unit in the World Health Organisation (WHO) South-East Asia Regional Office (SEARO) in New Delhi (India), the Western Pacific Regional Office (WPRO) in Manila (Philippines) or the Eastern Mediterranean Regional Office (EMRO) in Alexandria (Egypt). However, these offices do support a number of activities on zoonoses and food-borne diseases in WHO member countries of the region. Maintenance of the health of farmers and of their families (often termed "rural health") has assumed increasing importance in most member countries of the region. In most of the countries, there is no actual veterinary public health unit functioning as a national body common to the ministries of health and agriculture. Among the commonest zoonotic diseases prevalent in member countries are rabies, brucellosis, Japanese encephalitis, echinococcosis, tuberculosis, visceral leishmaniasis, taeniasis, salmonellosis, campylobacteriosis and leptospirosis. A national plan is necessary for each country to give priority to controlling these diseases, based on health systems research or primary health care, with intersectoral and regional cooperation through the South Asian Association for Regional Cooperation (SAARC) under Technical Cooperation among Developing Countries (TCDC). There should be a strong unit for veterinary public health in all WHO regional offices to coordinate zoonotic disease surveillance, training and control programmes in countries of the region.

Accessibility of dog populations for rabies control in Kathmandu valley, Nepal.

The accessibility of dogs in urban areas of Kathmandu valley was measured using the following approaches: determination of the proportion of dogs that bore signs of having been the objects of religious worship and other signs of household association, supplemented by information obtained by interviewing people in the neighbourhood; and the vaccination coverage attained in a rabies control campaign that was preceded by intensive activities to encourage the community to participate. An accessibility rate of 90-95% was determined using the first of these approaches, whereas 75-80% of the total dog population was reached in the vaccination campaign.