Interleukin-27 Disrupts the Crosstalk of Apoptotic Activities between 4T1 Breast Cancer Cells and M2 Macrophages

@#Introduction: Cytokine immunotherapy such as Interleukin-27 (IL-27) has been foreseen as a promising alternative anti-cancer treatment. Thus, this study aimed to investigate whether IL-27 gene therapy regulates crosstalk between breast cancer cells and macrophages in the sense of pro-apoptotic activities. Methods: This study has led to the development of recombinant pcDNA3.4-IL27. The recombinant pcDNA3.4-IL27 was transfected into 4T1 murine mammary carcinoma cells alone and co-culture of 4T1 with M2 macrophages. The successful expression of IL-27 in the cells were determine through the immunofluorescence staining and detection of CD206, M2 macrophages marker. Apoptotic effects of pcDNA3.4-IL27 were assessed through MTT assay, Annexin V flow cytometer analysis, and AO/PI dual staining. Results: Our findings shows that pcDNA3.4-IL27 has the ability to induce apoptosis in both of the cell group and performs better in the co-culture of 4T1 with M2 macrophages compared to 4T1 cells alone. PcDNA3.4-IL27 induced apoptosis through the altered cell morphology and reduction in the number of viable cells. Conclusion: These data demonstrate that pcDNA3.4-IL27 has the ability to induce apoptosis in both 4T1 cell alone and co-cultured 4T1 with M2 macrophages. Thus, could serve as a potential anti cancer candidate against breast cancer.

Evaluation of pancytopenia on bone marrow aspiration- study at a tertiary care center in Kashmir valley, India

Background: Pancytopenia refers to combination of anaemia, leukopenia and thrombocytopenia. It may be a manifestation of a wide variety of disorders, which primarily or secondarily affect the bone marrow. However, aetiology of pancytopenia varies from one geographical region to another. Bone marrow aspiration plays an important role in identifying the cause of pancytopenia. This study was carried to identify the causes of pancytopenia and to find out the bone marrow morphology in cases of pancytopenia.Methods: This study was conducted in the department of haematology in a tertiary care center in Kashmir valley for a period of 3 years. Inclusion criteria: cases with hemoglobin less than 10 gm/dl, total leucocyte count of less than 4000/mm3 and platelet count less than 100,000/mm3 were included in the study.Exclusion criteria: Patients receiving chemotherapy/radiotherapy were excluded from the study. Bone marrow aspiration (BMA) was performed from posterior iliac crest of the patients. Bone marrow aspiration smears were stained with Leishman stain for microscopy.Results: A total of 334 cases were studied during a period of 3 years. Age of patients ranged from 1 year to 85 years with mean of 43.59 years.180 cases were male, and 154 cases were female with male:female ratio of 1.2 :1. The commonest cause of pancytopenia was megaloblastic anemia seen in 103 cases (30.8%) followed by dual deficiency anemia seen in 69 cases (20.7%).37 cases (11%) were of acute leukaemia. Aplastic anemia was seen in 35 cases(10.5%). Other causes of pancytopenia were myelodysplastic syndrome, multiple myeloma, iron deficiency anemia and hypersplenism.Conclusions: Bone marrow aspiration in patients of pancytopenia helps in the identification of the underlying cause in most of the cases. BMA is helpful for understanding the disease process; and in planning further investigations and management of cytopenia patients.

Cytotoxicity, Regulation Of Apoptotic And Anti-Apoptotic Gene Expression By Il-27 In Mcf-7 And Mda-Mb-231 Breast Cancer Cell Lines / Jurnal Sains Kesihatan Malaysia

Breast cancer is one of the commonest cancers among women. Conventional therapies cause adverse side effects inpatients. Cytokine immunotherapy such as interleukin-27 (IL-27) has been sought as an alternative cancer treatment inrecent years. IL-27 has been shown to improve anticancer immunity and anti-angiogenesis in cancers, however, its effecton apoptotic and anti-apoptotic gene expression especially in breast cancers is yet to be explored. Cytotoxicity of IL-27in non-cancerous (184b5) and cancerous (MCF-7 and MDA-MB-231) breast cell lines was first determined for 24-72h in this study. The results indicated that IL-27 treatment did not retard 184b5 cell growth, however, did inhibit MCF-7(48 h) and MDA-MB-231 (72 h) cell growth with IC50 at 442 and 457 ng/ml, respectively. Apoptotic (TRAIL, FADD, FAS,caspase-3 and caspase-8) and anti-apoptotic (BCL-2, AKT, and COX-2) genes were then amplified from untreated (control)and treated breast cancer cells and studied. TRAIL, caspase-3, caspase-8 gene expression was significantly (p < 0.05)upregulated in treated MCF-7 (442 ng/ml) and MDA-MB-231 (457 ng/ml) cells. Expression of FADD and FAS genes wasnot detected in both control and treated MCF-7 and MDA-MB-231 cells. COX-2 gene was also not expressed by MCF-7cells, but reduced significantly (p < 0.05) in treated MDA-MB-231 cells. In MDA-MB-231 cells, IL-27 treatment seemedto slightly enhance the expression of AKT and BCL-2 genes which, on the other hand, was downregulated in treatedMCF-7 cells. Conclusively, IL-27 is able to inhibit breast cancer cell growth and regulate apoptotic and anti-apoptoticgene expression in breast cancer cells.

Phytochemical and biological assessments on Lipidium meyenii [maca] and Epimidium sagittatum [horny goat weed]

The effects of Lipidium meyenii [maca, LM] and Epimidium sagittatum [horny goat weed, ES] have been investigated due to their involvement in fertilization. Both of the drugs showed good results before, during and after fertilization in male and female mice. The results revealed that the crude extract of Lipidium meyenii caused a significant decrease in the no. of writhes at 300 and 500mg/kg [p<0.05] as compare to control, Epimidium sagittatum and standard drug. The gross behavioral, open field, exploratory behaviour, forced swimming test for stress, diuretic activity, chronic toxicity with the effect on reproduction of both male and female and change in body weight were also studied. The phytochemical study showed the presence of tannin, alkaloid, carbohydrate, rich protein and absence of sterol in LM, whereas ES shows presence of sterol and less protein. LS improve in muscle activity and exploratory behaviours without any toxic effects on mice and their pups. It does not have diuretic effect for first two hour but act normally after initial phase of drug therapy. Epimidium sagittatum has dual action that is at low dose it has slight stimulation action and at high dose little depressive effect. ES also has some diuretic effect. Overall these results suggest that LM is highly effective remedy for treatment of impotency and reduces stress and depression, because of dual effect ES not only suggested as an anxiolytic medicine but also effective in female hormonal disorder

Treatment Modalities for Skeletal Class III Malocclusion: Early to Late Treatment.

Skeletal Class III malocclusion has always been characterized by an unpredictable growth pattern with doubtful prognosis though treated at a very young age. Treatment protocols may include growth modification, orthodontic camouflage, or orthognathic surgery, depending on the degree of the skeletal discrepancy and the age of the patient. The orthopedic approach for growth modification is limited to the children in a growth phase. In a patient with underdeveloped maxilla, the face mask may be the treatment of choice, whereas in a Class III patient with over developed mandible a chin cap is needed. Camouflage orthodontic treatment is performed in patients with a mild skeletal Class III discrepancy and no remaining growth. However, adult patients with severe skeletal Class III discrepancy might need orthognathic surgery to correct the malocclusion.

Cytotoxic effect of proteinaceous postbiotic metabolites produced by Lactobacillus plantarum I-UL4 cultivated in different media composition on MCF-7 breast cancer cell

Aims: Lactobacillus sp. has capability of producing an array of bioactive metabolites that exhibit probiotic effects. Therefore, the objective of this study was to determine the cytotoxicity effect of proteinaceous postbiotic metabolites (PPM) produced by Lactobacillus plantarum I-UL4 cultivated in different media composition on MCF-7 breast cancer cell. Methodology and results: L. plantarum I-UL4 was cultivated in yeast extract and modified de Man, Rogosa and Sharpe broth containing Tween 80 (CRMRS+T80) or without Tween-80 (CRMRS-T80). Human breast adenocarcinoma cell (MCF-7) was employed as cancer cell in this study. Cytotoxicity and antiproliferative effects of PPM were determined using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl Tetrazolium Bromide assay and Trypan Blue Dye Exclusion assay, whereas Acridine Orange/Propidium Iodide staining was employed to determine the cytotoxicity mechanism. PPM produced in CRMRS+T80 exerted cytotoxicity in a time and dose dependent manner that was selective towards MCF-7 cancer cell. Profound cytotoxicity with the lowest IC50 concentration of 10.83 µg was detected at 72 h of incubation, whereas the most potent antiproliferative effect revealed by the lowest viable cell population was observed at 24 h of incubation. PPM cultivated in CRMRS+T80 induced 80.87% of apoptotic MCF-7 cells at 48 h of incubation. Conclusion, significance and impact of study: PPM of L. plantarum I-UL4 cultivated in different media composition induced different levels of MCF-7 cancer cell death. The percentage of apoptotic MCF-7 cells treated with PPM cultivated in CRMRS+T80 increased significantly (p < 0.05) from 24 to 48 h of incubation. The results obtained in this study have revealed the potential of PPM produced by L. plantarum I-UL4 as human health supplement and as anticancer preventive agent. Keywords: Lactobacillus plantarum I-UL4; cytotoxic effect; proteinaceous postbiotic metabolites; media composition; breast cancer

In vitro antioxidant and in vivo hepatoprotective effect on ethanol mediated liver damage of spray dried Vernonia amygdalina water extract

Vernonia amygdalina is a strong natural antioxidant that possessed various medicinal properties. In this study, the spray-dried water extract of V. amygdalina was evaluated for its in vitro antioxidant capacity and in vivo hepatoprotective effect against alcoholic-mediated liver damage. Total phenolic and flavonoid content of spray-dried V. amygdalina water extract were determined. Liver enzyme profiles, liver antioxidant level and nitric oxide level were evaluated in alcohol-induced liver injured mice or co-supplement with spray-dried V. amydalina. Water extract of spraydried V. amygalina that contained phenolic content of 24.8 +/- 1.5 mg/g gallic acid equivalent and total flavonoid content of 25.7 +/- 1.3 mg/g catechin equivalent was able to inhibit 50% of xanthine and tyrosinase oxidation at 170 micro g/mL and 2 mg/mL, respectively. On the other hand, extracts at both 10 and 50 mg/kg body weight were able to reduce the levels of Alanine transaminase [ALT], Alkaline phosphatase [ALP], Aspartate transaminase [AST], triglyceride and total bilirubin content inthe alcohol-mediated liver injury in mice. Furthermore, it also helped to increase levels of Superoxide dismutase [SOD], Ferric reducing ability of plasma [FRAP] and reduce the levels of Nitric oxide [NO] and Malondialdehyde [MDA] in the liver of the treated mice. These results suggested that water extract of spray-dried V. amygdalina exhibited liver protective effect, which could be contributed by its antioxidant properties

Maternal and fetal outcomes in rheumatic heart disease in pregnancy.

Background: Rheumatic Heart Disease remains the most common type of heart disease in pregnancy in developing countries. Over a period of 2 years,129 pregnant patients with Rheumatic Heart Disease attending our outpatient department, were observed from admission till discharge and the clinical course during pregnancy and the maternal, foetal outcomes, and desire of future contraception studied. Methods: Patients were routinely examined every antenatal visit for signs of anemia and congestive cardiac failure. They were usually admitted at 28-32 weeks of gestation unless they presented with symptoms of cardiac failure earlier in pregnancy. Frequency of the antenatal visits depended on the functional cardiac status. Results: The incidence of heart disease was 0.96% for all deliveries during our study period. The incidence of RHD was 71.6%. 65.1% of women gave a h/o rheumatic fever. 45(34.9%) patients had undergone surgical correction and 84(65.1%) did not undergo surgical correction. 95.3% had moderate to severe disease but only 18.6% were in NYHA class III/IV. Hence severity of disease did not correlate with NHYA class. 76.7% of women had vaginal delivery. LSCS was done for obstetric reasons alone. Conclusion: The association of the pre pregnancy functional class with the risk of maternal events raises attention to the possibility of reducing these complications in pregnant women with mitral stenosis by means of early interventions aimed at improving their functional class.

To evaluate the effect of yoga on moderate degree hypertension and lipid profile.

Yoga is a practice consisting of gentle stretching, breath control and mind-body intervention. So we aimed at yoga practice on blood pressure and lipid profile. Methods: Carried out at Dept of Cardiology, Owaisi Hospital and research centre. After taking written informed consent, we selected 60 Patients (30 females and 30 males) age 40 years to 60 years who attended our programs with moderate hypertension who is volunteered to participate in the study. They were trained in asanas (postures), Pranayama (breathing exercise) and relaxation techniques for 6 months. Blood pressure, Serum total cholesterol, LDL, VLDL, HDL cholesterol and total triglycerides were measured at the beginning (baseline, day 0) and at the end of the study (after 6 months). Results: The systolic blood pressure came down from 164.0+ 1.9 to 140+1.9 mmHg, Diastolic blood pressure 96+0.8 to 82+0.6 mmHg, pulse rate declined from 85+1.2 to 77 + 0.7 per min, the TC concentration decreased significantly from 200+ 6.5 to 170+ 3.6 mg/dl (p< 0.001) the LDL reduced from 166+ 4.2 to 148+3.7 mg/dl (p<0.001) and the triglycerides showed a significant decrease from 189+10.3 to 166+8.6 mg/dl (p<0.001), while the HDL cholesterol showed a marked increase from 39+1.4 to 46+1.2 mg/dl (p<0.05). Conclusion: The results conclude that the Yoga practice in patients with moderate degree hypertension leads to decrease in blood pressure and lipid profile within the period of 6 months.

Status of heavy metals distribution in municipal solid waste in Tiruchirappalli City, India.

Urbanization and rapid growth of population in India has led to drastic increase in municipal solid waste. Unscientific disposal of municipal solid waste is one of the main reasons attributed for environmental degradation. The present work concentrates on municipal solid waste management in Tiruchirappalli City which comprises of four zones namely Srirangam, Goldenrock, Araiyamangalam and Abishekapuram. This study also attempted to assess the physical composition, characteristics and the heavy metals content in municipal solid waste. It can be observed that the bio-degradable fraction of municipal solid waste is found to be 74 percent of the total solid waste generated from the city. Hence composting could be the best option for the treatment of municipal solid waste.

Sybr Green Real-Time PCR is a Useful Method in Screening for High Risk Human Papilloma Virus Types 16 and 18 in Neoplastic Cervical Cancer

Human papillomavirus (HPV) plays an important role in the pathogenesis of cervical cancer. HPV has been found in 99.7% of cervical cancers worldwide. In Malaysia, it is the second most common cancer among women in all major ethnic groups. The main purpose of this study was to establish the method of SyBrGreen Real-Time PCR and apply it for identification of multiple infections of the two high risk HPV subtypes. In this study, 57 positive samples for HPV 16 and HPV 18 were used to establish a simple and sensitive method to detect and identify HPV infection in the cervical neoplasia at different stages of the disease by using real-time ABICycler SyBrGreen 1 technology. The results showed 67 HPV genomes in 57 samples. HPV 16 genome was detected in 55/67 (82%) cases while HPV 18 was detected in 8/67 (12%) cases with 4 cases showing multiple infections of HPV 16 and HPV 18. HPV 16 was the most prevalent followed by HPV 18. Using SyBr Green Real-Time PCR techniques, the results showed that DNA melting curve for HPV 16 had a peak around 80.2ºC and Ct value of 20 cycles whereas the DNA melting curve for HPV 18 around 79.2ºC and Ct INTRODUCTION value of 22 cycles. In conclusion, a SyBr Green Real-Time PCR method has the potential for clinical usage in detection and identification of HPV infection in cervical neoplasia at different stages of the disease.

Cytotoxic effects of commercial wheatgrass and fiber towards human acute promyelocytic leukemia cells [Hl60]

Cytotoxicity, the possible selective activity upon HL60 as well as the anti-proliferation effect of local health supplement wheatgrass and mixture of fibers were investigated in vitro using various cancerous cell line and normal blood cell culture. The IC[50] of wheatgrass-treated HL60 [17.5 +/- 1.1, 12.5 +/- 0.3, and 16 +/- 0.5 microgram/ml for 24, 48 and 72 h, respectively] and fibers-treated HL60 [86.0 +/- 5.5, 35.0 +/- 2.5, and 52.5 +/- 4.5 microgram/ml for 24, 48 and 72 h, respectively] showed that both extracts possessed optimum effect after 48 hours of treatment. No significant cytotoxic effect was observed on other type of cells. For trypan blue dye exclusion method, wheatgrass reduced the number of viable cells by 13.5% [ +/- 1.5], 47.1% [ +/- 3.6], and 64.9% [ +/- 2.7] after 24, 48 and 72 h exposure, respectively. Mixture of fibers reduced the number of viable cells by 36.4% [ +/- 2.3], 57.1% [ +/- 3.1], and 89.0% [ +/- 3.4] after 24, 48 and 72 h exposure, respectively, indicated that necrosis is also an alternative to the apoptotic mechanism of cell death. Annexin-V/propidium iodide staining revealed that both extracts induced apoptosis where early apoptosis had been detected concurrently with the reduction of percentage of cell viability. Cell cycle analysis revealed that in HL60, the percentage of apoptosis increased with time [wheatgrass: 16.0% +/- 2.4, 45.3% +/- 3.4 and 39.6% +/- 4.1; mixture of fibers: 14.6% +/- 1.8, 45.4% +/- 2.3 and 45.9% +/- 1.2] after exposure for 24, 48 and 72 h, respectively at the concentration of 100 microgram/ml and showed optimum effect at 48 hours. Thus, these health products can be a potential alternative supplement for leukaemia patients

Immunomodulatory effects of zerumbone isolated from roots of zingiber zerumbet

In this study, the immunomodulatory effects of zerumbone isolated from Zingiber zerumbet were investigated by evaluating the effects of this compound towards the lymphocytes proliferation [mice thymocytes, mice splenocytes and human human peripheral blood mononuclear cells, PBMC], cell cycle progression and cytokirie [interleukin 2 and 12] induction. Lymphocyte proliferation assay showed that zerumbone was able to activate mice thymocytes, splenocytes and PBMC at dosage dependent pattern where the best concentration was 7.5 micro g/mL. Flow cytometry analysis showed the highest population of PBMC entered into G2/M phase after treatment for 72 h with 7.5 micro g/mL zerumbone. The production of human interleukin-2 and human interleukin-12 cytokines in culture supernatant from zerumbone activated lymphocytes was prominently upregulated at 24 hour and decreased from 48 h to 72 h. The above results indicate that zerumbone can be used as immunomodulatory agent which can react toward the immune cell cytokine production in dosage dependent pattern

Gestational diabetes mellitus-screening in a developing country

All gravid women attending the antenatal clinic of the department of Gynaecology Unit-II, Civil Hospital Karachi and unbooked cases presenting in labour from October, 1986 to December, 1993, were included in the study. On the basis of risk factors for diabetes mellitus [DM], they were divided into 3 groups. Women in Group-I had family history of DM, bad obstetric history, congenitally malformed babies, polyhydramnios, obesity and macrosomia. Group-II included women with pregnancy induced hypertension [PIH] abruptio placentae, age >30 years and multiple pregnancy. Group-Ill comprised of the pregnant women with no risk factors for DM. Screening of 4497 subjects from the antenatal clinic was performed and DM was diagnosed in 211[12%] in Group-I, 11[3%] in Group-Il and 13[0.6%] in Group III. Chi-square analysis demonstrated that frequency of DM was significantly higher [P<0.001] in Group I and Group II as compared to Group III. Non- booked admissions numbered 7600, of these, DM was detected in 166[20%] belonging to Groups-I and II, whereas, 38 [0.6%] were from Group-Ill. Screening was done by the O'Sullivan criteria. It was concluded that all pregnant women should be screened for DM. However, if financial constraints prevail, then women with high risk factors only should be subjected to screening