Protease inhibitors as a potential agent against visceral Leishmaniasis: A review to inspire future study

Abstract Leishmaniasis is transmitted by sandfly which carries the intracellular protozoa in their midgut. Among visceral, cutaneous and mucocutaneous leishmaniasis, visceral type that is caused by Leishmania donovani is the most lethal one. Findings of leishmanial structure and species took place in 19th century and was initiated by Donovan. Leishmaniasis is still a major concern of health issues in many endemic countries in Asia, Africa, the Americas, and the Mediterranean region. Worldwide1.5-2 million new cases of cutaneous leishmaniasis and 500,000 cases of visceral leishmaniasis are reported each year. Leishmaniasis is endemic in nearly 90 countries worldwide and close to 12 million new cases of leishmaniasis are reported worldwide annually. Studies on antileishmanial drug development is of major concern as leishmaniasis are the second largest parasitic killer in the world and the available drugs are either toxic or costly. The major surface GP63 protease, also known as Zinc- metalloproteases present on the surface of leishmanial promastigotes, can be targeted for drug development. Protease inhibitors targeting such surface proteases show promising results. Different protease inhibitors have been isolated from marine actinobacteria against many infectious diseases. Metabolites produced by these actinobacteria may have greater importance for the discovery and development of new antileishmanial drugs. Hence, this review discusses the background, current situation, treatment, and protease inhibitors from marine actinobacteria for drug development against GP63 molecules.

Evaluation of Serum Transaminases in Type 2 Diabetes Mellitus

Background Liver is the central hub for metabolism. Liver dysfunction in diabetes mellitus is one of the major causes of morbidity and mortality. Periodical evaluation of transaminases helps in early diagnosis of liver dysfunction. Aim The aim of the present study is to measure aspartate transaminase (AST) and alanine transaminase (ALT) in known cases of type 2 diabetes mellitus (T2DM) and to compare the values with matched controls. Settings and Design Institutional cross-sectional observation study. Methods & Material Study was done in 100 known cases of T2DM and in 30 controls. Age, AST, ALT and fasting plasma glucose (FPG) were recorded, analysed and compared between two groups. Statistical Analysis Data was analysed using Microsoft Excel 2007 and SPSS trial version 16.0. Results Significant difference between FPG, AST, ALT and age were observed between two groups (P < 0.05). Conclusion The results from our study showed that there are elevated levels of ALT and ASTs among T2DM patients when compared with normal individuals.

Reactive Plasmacytosis Associated with Chronic Generalized Periodontitis – A Case Report.

Plasmacytosis is a condition in which there are an unusually large proportion of plasma cells in tissues, exudates, or blood. Reactive plasmacytosis is a diagnosis of exclusion, to be differentiated from other autoimmune, allergic and neoplastic disorders with plasma cell infiltrates. This paper describes a rare case of 73-year-old male patient diagnosed as plasmocytosis depending upon the clinical and histological findings. There was chronic inflammatory enlargement of the gingiva and palate with severe periodontitis. Histopathological examination revealed a stratified squamous orthokeratinized epithelium with underlying fibrocellular connective tissue stroma. The connective tissue showed intense infiltrate of small round cells of plasmacytoid type with eccentrically placed nucleus suugestive of reactive plasmacytosis.The diagnostic and management challenges encountered are described in this case report.

Profile of HIV positive clients in an ICTC of a private medical college, Andhra Pradesh: A situational analysis.

Background: HIV/AIDS has turned out into global pandemic. In India under National AIDS Control Program (NACP) –III Integrated Counseling and Testing Centers (ICTC) were established throughout the country for providing prevention services through a holistic approach for all segments of the population. Objectives: To study the sociodemographic profile and risk behavior pattern of clients diagnosed HIV positive in the ICTC. Material & method: Study Design: Cross-sectional study (record based) Setting: Mediciti Institute of Medical Sciences, Andhra Pradesh. Subjects- All the 448 seropositive attendees during the year 2007-2010. Data pertaining to total 8845 clients, who availed the services from the ICTC during the above period was assessed for the time trend of HIV infection .Study Variables- Year of reporting, in-referral of the clients, age, sex, educational status, marital status, occupation and pattern of risk behavior in relation to HIV/AIDS. Statistical analysis- Data was analyzed in SPSS version 17.0 . Results: HIV prevalence was found to be declining over the last 4 years, except in 2008. Among the seropositives 62.7% were males and 37.3% were females. 85% were in the reproductive age group 78.1% were married;53.1% were illiterates. Majority of them are labourers from the agricultural background and housewives. In 83.9% of the individuals it is acquired through unprotected heterosexual intercourse. Conclusion- These findings should call for greater introspection among the program managers and facilitate for more customized local planning involving various stakeholders.

Iatrogenic hemorrhage from left umbilical vein after cardiac surgery: An unusual complication.

Postoperative bleeding is a concern for all patients undergoing heart surgery, which could be due to surgical causes or coagulation disorder. The patients at risk for coagulopathy include those patients with complex or prolonged procedures, those exposed to preoperative anticoagulants and, to a lesser extent, patients with a preoperatively elevated prothrombin time and activated clotting time. However, intraabdominal bleeding after cardiac surgery is rare (0.3-2%). As the mortality rate of patients exposed to these complications is high (11-59%), timely recognition and prompt management is vital for patient's safety and for avoidance of postoperative complications. Here, we present a case of free intraabdominal hemorrhage as sequelae of pacing wire insertion in open heart surgery and its successful management.

Aesthetic Crown Lengthening.

Aesthetics is an inseparable part of today’s dental treatment and has influenced the management of dental maladies in varying degrees for many years. Patient awareness and expectations have increased recently to the point that less than optimal aesthetics are no longer an acceptable outcome. Achieving an appealing and an aesthetic smile is no longer a dream. In this article an attempt has been made to review the guidelines for treating a patient with excessive gingival display along with gingival pigmentation and its treatment modalities with illustrations from two case reports.

Assessment of long term status of sputum positive pulmonary TB patients successfully treated with short course chemotherapy.

Background: Long term status of pulmonary tuberculosis (PTB) patients treated with short course chemotherapy (SCC) regimens remains unknown. Objective: To assess the clinical, bacteriological, radiological status and health related quality of life (HRQoL) of PTB patients 14 -18 years after successful treatment with SCC. Methodology: In a cross-sectional study, cured PTB patients treated during 1986 – 1990 at the Tuberculosis Research Centre (TRC) were investigated for their current health status including pulmonary function tests (PFT). The St Georges respiratory questionnaire (SGRQ) was used to assess the HRQoL Results: The mean period after treatment completion for the 363 eligible participants was 16.5yrs (range 14-18 yrs, 84% coverage) ; 25 (7 %) had been re-treated and 52 (14%) died. Among the investigated, 58 (29%) had persistent respiratory symptoms; 170(86%) had radiological sequelae but none had active disease. Abnormal PFT was observed in 96 (65%) with predominantly restrictive type of disease in 66(45%). The SGRQ scores for activity and impact were high implying impairment in HRQoL. Conclusion: Assessment of long term status of cured PTB patients showed an impairment of lung functions and HRQoL highlighting the need to address these issues in the management of TB that may provide added value to patient care.

Paullinia cupana Mart var. sorbilis, guaraná, reduces cell proliferation and increases apoptosis of B16/F10 melanoma lung metastases in mice

We showed that guaraná (Paullinia cupana Mart var. sorbilis) had a chemopreventive effect on mouse hepatocarcinogenesis and reduced diethylnitrosamine-induced DNA damage. In the present experiment, we evaluated the effects of guaraná in an experimental metastasis model. Cultured B16/F10 melanoma cells (5 x 10(5) cells/animal) were injected into the tail vein of mice on the 7th day of guaraná treatment (2.0 mg P. cupana/g body weight, per gavage) and the animals were treated with guaraná daily up to 14 days until euthanasia (total treatment time: 21 days). Lung sections were obtained for morphometric analysis, apoptotic bodies were counted to calculate the apoptotic index and proliferating cell nuclear antigen-positive cells were counted to determine the proliferation index. Guaraná-treated (GUA) animals presented a 68.6 percent reduction in tumor burden area compared to control (CO) animals which were not treated with guaraná (CO: 0.84 ± 0.26, N = 6; GUA: 0.27 ± 0.24, N = 6; P = 0.0043), a 57.9 percent reduction in tumor proliferation index (CO: 23.75 ± 20.54, N = 6; GUA: 9.99 ± 3.93, N = 6; P = 0.026) and a 4.85-fold increase in apoptotic index (CO: 66.95 ± 22.95, N = 6; GUA: 324.37 ± 266.74 AB/mm², N = 6; P = 0.0152). In this mouse model, guaraná treatment decreased proliferation and increased apoptosis of tumor cells, consequently reducing the tumor burden area. We are currently investigating the molecular pathways of the effects of guaraná in cultured melanoma cells, regarding principally the cell cycle inhibitors and cyclins.

High intensity exercise training programme following cardiac transplant.

A 26-year-old male patient who presented with symptoms of end stage cardiac failure as a result of dilated cardiomyopathy, had an orthotopic cardiac transplantation. A comprehensive cardiac rehabilitation programme was provided to him and he was introduced to a sport (tennis). The exercise training programme progressed from low intensity training to high intensity programme over a period of 15 months. A cardio-pulmonary exercise test done 22 months after surgery suggested that he was able to achieve the aerobic capacity comparable to that of a normal South Indian subject. He participated successfully in the World Transplant Games in Sydney and returned safely. This suggests that after a proper cardiac rehabilitation programme, patients undergoing heart transplantation can achieve normal physiological responses to lead a normal active life.

Differential role of MAP kinase isoforms in malachite green transformed Syrian hamster embryo fibroblasts in culture.

Malachite green (MG) induces DNA damage and malignant transformation of Syrian hamster embryo (SHE) cells in primary culture. In the present study, we have studied the role of all the three isoforms of mitogen activated protein (MAP) kinases i.e. ERK (extracellular regulated kinase), JNK (JUN- N- terminal kinase) and p38 kinase during transformation of SHE cells by MG. The results showed that transformed cells were associated with a decreased expression of phosphoactive ERK and JNK and increased expression of p38 kinase as evident from the Western blot, immunofluorescence and flow cytometry studies. Also, a persistent nuclear localization of p38 kinase was observed in the transformed cells. The present study indicated that p38 kinase was present at higher levels and seemed to be associated with transformation, which suggested that inhibitors of p38 kinase could serve in general as potential agents for selective cancer therapy.

Inhibitory effect of cinnamoyl compounds against human malignant cell line.

In the present study, anti-proliferative effects of dietary polyphenolic compounds have been observed and demonstrated the strong anticancer efficacy of curcumin (CMN), an active constituent of dietary spice (turmeric) using human leukemia cancer cell line. CMN inhibited the proliferation of K562 leukemic cells by induction of apoptosis. The current study demonstrated synergy with combination of drug therapy, and suggested that combination of ferulic acid and cisplatin synergistically inhibited cellular proliferation. Cytotoxic synergy was observed independent of the sequence of addition of two drugs to cultured cells. The synergized growth inhibitory effect with cisplatin was probably associated with G2-M arrest in cell cycle progression. These findings suggested that among the cinnamoyl compounds, CMN was most potent and FER appeared to be a better modulating agent on human malignant cell line.

Chemopreventive effect of orange oil on the development of hepatic preneoplastic lesions induced by N-nitrosodiethylamine in rats: an ultrastructural study.

Orange peel oil is used extensively as an approved flavour enhancer in fruit drinks, carbonated beverages and as a scenting agent in soaps and cosmetics. Limonene, which is a monocyclic monoterpene is present in orange peel oil from 90 to 95% (w/w). Monoterpenes have been shown to be very effective chemopreventive agents against several rodent tumors and are currently in clinical trials. However, not much information is available regarding the ultrastructural changes associated with the chemopreventive effects of the monoterpenes. The effect of orange oil on the suppression of preneoplastic hepatic lesions during N-nitrosodiethylamine (DEN) induced hepatocarcinogenesis was studied electron microscopically. Rats were administered 200 ppm DEN through drinking water for a period of 1 month. After an interval of 2 weeks, the animals were administered orange oil by gavage for a period of 5 1/2 months. The chemopreventive effect of orange oil was monitored on the basis of liver weight profile, histological pattern by light microscopy and ultrastructural alterations by electronmicroscopy. Orange oil administration following DEN treatment showed decreased liver weights, increased intercellular gap junctional complexes, cell density and polarity when compared with only the DEN treated rats. In the present study chemopreventive effect of orange oil on DEN-induced hepatic preneoplasia in rats which is associated with the restoration of the normal phenotype and upregulation of junctional complexes has been demonstrated.

Effect of metanil yellow and malachite green on DNA synthesis in N-nitrosodiethylamine induced preneoplastic rat livers.

Metanil yellow (MY) and malachite green (MG) are textile dyes, which, despite the ban occurs unsrupulously as food colouring agents. Accordingly they constitute a serious public health hazard and are of sufficient environmental concern. We have earlier reported that both MY and MG have tumor enhancing effects on the development of hepatic preneoplastic lesions induced by N-nitrosodiethylamine in rats. In order to understand the possible mechanisms by which MY and MG enhance tumor development, in this study we have tested the effects of MY and MG on DNA synthesis and PCNA expression in preneoplastic hepatic lesions during N-nitrosodiethylamine (DEN) induced hepatocarcinogenesis in male Wistar (WR) rats. Rats were administered 200 ppm DEN through drinking water for a period of one month. Administration of DEN for a period of one month showed an upregulation of cell cycle regulatory proteins namely cyclin D1, CDK4, cyclin E and CDK2. Accordingly, in other experiments, the animals were further administered MY and MG for a period of one month following one month DEN treatment. The effects of MY and MG were monitored on the basis of cell proliferation markers--DNA synthesis and PCNA expression both by immunohistochemical and immunoblotting. Following DEN administration, MY, MG and PB showed stimulation of DNA synthesis and increased PCNA expression when compared with either the corresponding controls or only DEN treated animals. In the present study, enhancing effect of MY, MG and PB on the cell proliferation markers during DEN-induced hepatic preneoplasia in rats was observed.

Identification of an appropriate strategy to control anemia in adolescent girls of poor communities.

OBJECTIVES: To obtain baseline data on hemoglobin (Hb) levels of adolescent girls belonging to the low-socio-economic groups; investigate the comparative efficacy of once 'weekly' and 'daily' administration of iron-folate tablets with respect to impact on the Hb levels; and find out the effect of added ascorbic acid supplementation on the efficacy of iron-folate administration with respect to increment in Hb levels. DESIGN: Randomized experimental. SETTING: Adolescent girls of poor communities in urban areas of Delhi and rural parts of Bharatpur (Rajasthan). METHODS: The baseline investigations included measurements of height, weight, and Hb levels. The Hb levels of the participating subjects were measured again after 3 months and 6 months of supplementation. RESULTS: 61.9% of the subjects in the urban and 85.4% in the rural area were anemic. The response of Hb levels to daily iron/folate supplementation was better in comparison to once-weekly supplementation. The increment in Hb levels of subjects due to addition of vitamin C to iron/folate supplementation was more than that with supplementation of iron/folate alone. CONCLUSIONS: Considering compliance, feasibility and cost-factors, a public-health approach consisting of once-weekly distribution of iron/folate supplementation through schools and welfare centers is better and can be recommended as an appropriate strategy for combating anemia in adolescent girls of poor communities in developing countries like India.

Malignant transformation of Syrian hamster embryo (SHE) cells in culture by malachite green: an agent of environmental importance.

Malachite green (MG), consisting of green crystals with a metallic lustre, is very soluble in water and is highly cytotoxic to mammalian cells in culture and also acts as a liver tumour promoter. In view of its industrial importance and possible exposure to human beings, MG poses a potential environmental health hazard. Accordingly, we have studied the effect of MG on the formation of free radicals using Electron Spin Resonance (ESR) analysis with 5,5-dimethyl-1-pyrroline N-oxide (DMPO) as a spin trapping agent. ESR analysis showed formation of reactive free radicals during exposure of MG to Syrian hamster embryo (SHE) cells. As per mechanism-based toxicology in cancer risk assessment, the chemicals that have the potential to be metabolized to active free radical species could be human cancer hazards. So, we have investigated the effect of MG on the formation of Type II and Type III morphologically transformed foci using SHE cell transformation assay. MG induced dose related transformed foci. Some of these transformed foci were taken out using selective trypsinisation and established immortal cell lines. One of these immortal cell lines was characterized extensively. This immortal cell line showed enhanced DNA synthesis in the form of BrdU incorporation, increased presence of proliferating cell nuclear antigen (PCNA), bcl-2 and p53 proteins by immunohistochemistry. When these immortal cells were injected subcutaneously into nude mice, they developed tumors which were transplantable and histopathologically sarcomas. The present studies indicate that MG could be a potential candidate for two year chemical carcinogenesis rodent bioassays.

Suitability of CANSCORE for the assessment of the nutritional status of newborns.

The suitability of Clinical Assessment of Nutritional Status Score (CANSCORE) for the assessment of foetal malnutrition among 372 local Hyderabad newborns was studied. Details of length, weight and body mass index (BMI) at birth were related to total CANSCORE which consisted of scores ranging from 1 to 4 based on the grades of clinical status of hair, cheeks, buttocks, chest, legs, back, neck, arms and skin of anterior abdominal wall. The correlation coefficients of CANSCORE with the length, weight and BMI of newborns indicated that score of hair was least correlated with nutritional status. Normal newborns were found to have the lowest prevalence of foetal malnutrition. In those with retarded measurements of length and weight or BMI, the prevalence of foetal malnutrition was higher. The newborns with retardation of both length and BMI had higher prevalence of foetal malnutrition. The feasibility of the suggested limits of CANSCORE for the foetal malnutrition was assessed. Values of 24 for total CANSCORE and of 22 for "Modified CANSCORE" (score excluding hair as a parameter) were found appropriate for the assessment of foetal malnutrition. Modified CANSCORE is a simple, rapid and quantifiable method for the assessment of foetal malnutrition in term newborns.

Sequential enhanced tyrosine phosphorylation during progressive malachite green induced malignant transformation of Syrian hamster embryo cells in culture is associated with no change in the activity levels of tyrosine phosphatases.

Malachite green (MG) consisting green crystals with a metallic lustre is extremely soluble in water and is highly cytotoxic to mammalian cells and also acts as liver tumor promoter. In view of its industrial importance and possible exposure to human beings, MG poses a potential environmental health hazard. We have earlier reported that MG induces malignant transformation in Syrian hamster embryo (SHE) cells. Since tyrosine phosphorylation and dephosphorylation reactions are known to play critical roles during normal and abnormal cellular proliferation, in this study we have studied the tyrosine phosphorylation, tyrosine phosphorylated proteins and protein tyrosine phosphatases in malignantly transformed cells and during sequential development of cellular transformation by MG compared to control cells. The present investigation shows that enhanced tyrosine phosphorylation and tyrosine phosphorylated proteins associated with the static levels of tyrosine protein phosphatases may probably contribute to the abnormal cellular proliferation during malignant transformation of SHE cells by MG.

Effect of dose and formulation on iron tolerance in pregnancy.

BACKGROUND: The National Nutritional Anaemia Prophylaxis Programme (NNAPP) in India was launched in 1971. However, anaemia continues to be a major public health problem. Partial coverage of the population, inadequate dose of the iron supplement, defective absorption due to intestinal infestations and problems with formulation have been recognized as factors responsible for its failure. Therefore, the bioavailability of iron from different formulations containing 60 mg of elemental iron and of tablets with varying doses of elemental iron was undertaken. METHODS: One hundred and fifteen women were randomly allotted to receive different formulations and doses of iron and then undergo iron tolerance tests. They received ferrous sulphate tablets containing 60 mg, 120 mg and 180 mg of elemental iron; formulations containing 60 mg of elemental iron as pure ferrous sulphate salt, ferrous fumarate tablets, ferrous fumarate syrup, excipients added to pure ferrous sulphate salts, powdered ferrous sulphate tablets, iron tablets distributed by the NNAPP and pure ferrous salt in gelatin capsules. RESULTS: The data obtained from 32 subjects were excluded because of non-compliance, intolerance of the medication and lack of results of blood tests. Data from the remaining 83 subjects indicated that increasing the dose of elemental iron from 60 mg to 180 mg improved the bioavailability of iron, but was associated with unacceptable side-effects. Also, liquid formulations of iron had a better bioavailability, with ferrous fumarate syrup and gelatin capsules being the most superior. CONCLUSION: Providing the iron formulation with a high bioavailability should enable the NNAPP to be more successful in decreasing the prevalence of anaemia.

Loss of a putative novel growth inhibitory apoptotic 14 kD polypeptide during progression of rat liver carcinogenesis.

Carcinogenesis is a multistep process involving different stages. However, the biological and biochemical factors responsible for the stepwise transition of cells from one stage to the other remains as important enigmas even today. We have recently isolated a putative novel growth inhibitory apoptotic 14 kD polypeptide from normal rat liver. In order to understand the possible functional relationship between 14 kD polypeptide and liver carcinogenesis, the sequential expression of this polypeptide as a function of tumor progression was studied in the rat liver using diethylnitrosamine (DEN) as a carcinogen. Immunoperoxidase and immunoblotting experiments using polyclonal rabbit antisera revealed a gradual reduction in the levels of this polypeptide with tumor progression. No reduction in the levels of this polypeptide was observed in regenerating rat liver after partial hepatectomy. The findings suggest that the loss or reduction of 14 kD polypeptide is linked selectively to abnormal cell proliferation and appears to be a biologically relevant risk factor for the progression of hepatocarcinogenesis in rats.