PubMed Health⌕ Search

Biomedical subjects

M Thakar

Publications and source records attributed to M Thakar.

16 recordsLinked to original sources
PubMed2003

Comparing modified and plain peptide linked enzyme immunosorbent assay (ELISA) for detection of human immunodeficiency virus type-1 (HIV-1) and type-2 (HIV-2) antibodies.

Serological diagnosis of human immunodeficiency virus (HIV) based on detection of HIV antibodies is one of the easiest, cheapest and simplest assay. Synthetic peptides corresponding to immunodominant regions of envelope glycoprotein (gp41, V3 loop for HIV-1 and gp36 for HIV-2) were used in the present study, to detect the anti-HIV antibodies in sera of Sexually Transmitted Diseases (STD), Tuberculosis (TB), Anti-Natal Care (ANC) patients. About 550 serum samples were tested using Enzyme Linked Immunosorbent Assay (ELISA) technique. The human sera positive for antibody to HIV-1 and HIV-2, reacted to different degrees with these peptides when used as a plain peptide with or without CGG motif/biotin motif at the amino terminus. The selected sequences are of Indian strain with 'C' serotype. The results showed a 100% sensitivity and specificity for V3 loop peptide and 98% sensitivity and specificity for gp41 peptide containing CGG moiety while the plain peptides showed similar sensitivities but low specificity's, i.e. 98% for V3 loop peptide and 42% for gp41 peptide when reacted with HIV-1 positive sera. The presence of biotin at the amino terminus did not provide any beneficial effect in increasing the sensitivity although the specificity was enhanced for both the peptide sequences, i.e. gp41 and V3 loop peptide. Furthermore, the gp36 peptide containing CGG moiety detected the HIV-2 sera with 100% sensitivity and 98% specificity while the sensitivity and specificity of gp36 plain peptide was reduced to 98 and 90%. Thus the study overall highlighted the importance of synthetic peptides containing CGG moiety as a capture antigen in detecting both HIV-1 & 2 sera using an indigenously built ELISA system which is simple, cheap, sensitive and cost effective for rural areas.

Enzyme-Linked Immunosorbent Assay↗
PubMed2001

Insulin-like growth factor type 1 (IGF-1) and IGF binding protein-3 in patients with Ewing sarcoma family of tumors.

BACKGROUND: Ewing sarcoma family of tumors (ESFTs) are the second most common bone tumor, that most often affects persons ages 3-40 years. The ESFTs rely on signaling through the insulin-like growth factor-1 receptor (IGF-1R) for growth and transformation. The current studies were performed to determine the levels of IGF-1 and IGF binding protein-3 (IGFBP-3) in patients with ESFT. The authors then performed an exploratory analysis to evaluate whether IGF parameters could differentiate event free or overall survival in ESFT patients. METHODS: The authors measured serum levels of IGF-1 and IGFBP-3 by using a radioimmunoassay from 111 patients with ESFT with a median follow-up of 13 years from diagnosis. RESULTS: The IGF-1 levels were lower among patients with metastatic disease to the bones or the bone marrow compared with patients without metastasis to these sites (p2 = 0.021 and 0.0038, respectively). IGFBP-3 is known to sequester IGF-1; the ratios of IGFBP-3 to IGF-1 were evaluated. Patients with metastatic disease to any site had higher IGFBP-3 to IGF-1 ratios than patients with localized disease (p2 = 0.0067). There was a trend toward increased survival in patients with localized disease who had high IGFBP-3 to IGF-1 levels. Metastatic patients showed a similar trend. CONCLUSIONS: Levels of IGF-1 and IGFBP-3 in ESFT patients can identify patients with the most widespread disease. The IGFBP-3 to IGF-1 ratio in patients with either localized or metastatic disease identified patients with a trend toward increased survival. Further prospective evaluation with higher patient numbers might show a prognostic role for the IGFBP-3 to IGF-1 ratio in patients with ESFT.

Biomarkers, Tumor↗
PubMed1999

Phosphoinositide 3-hydroxide kinase blockade enhances apoptosis in the Ewing's sarcoma family of tumors.

Ewing's sarcoma family of tumors (ESFTs) affects patients between the ages of 3 and 40 years, with most cases occurring in the second decade of life. These tumors contain a characteristic translocation, t(11;22), that produces a unique fusion protein, EWS/FLI-1. EWS/FLI-1 transforms mouse fibroblasts; this transformation requires intact EWS and FLI-1 domains as well as the insulin-like growth factor-I receptor (IGF-IR). The IGF-IR is a well-described transmembrane tyrosine kinase receptor that modulates transformation, cell growth, and survival. IGF-IR survival signaling is mediated through the downstream activation of phosphoinositide 3-OH kinase (PI 3-K) and Akt. Apoptosis, programmed cell death, progresses from a central death signal to a caspase cascade, including activation of caspase-3. Because the IGF-IR has been shown to play a role in the transformation and growth of ESFTs, we wanted to determine the role of downstream molecules in the cellular response to doxorubicin treatment. Doxorubicin increased caspase-3 activity in two ESFT cell lines, TC-32 and TC-71. Concomitant treatment of the doxorubicin-treated cells with IGF-I reduced caspase-3 activity 8-fold in TC-32 and 4-fold in TC-71. To determine whether PI 3-K has a role in IGF-I-mediated survival in ESFTs, PI 3-K was then inhibited with wortmannin and LY294002. Doxorubicin treatment reduced cell number and enhanced apoptosis in PI 3-K inhibited cells compared with noninhibited cells. Akt, a serine/threonine kinase activated downstream of PI 3-K, was investigated to determine whether its constitutive activation would render ESFT cells more resistant to doxorubicin. A constitutively activated Akt was stably transfected into ESFT and those cells with high levels of expression demonstrated increased resistance to doxorubicin-induced caspase-3 activation. These results indicate that ESFT cell lines use an IGF-IR initiated signaling pathway through PI 3-K and Akt for survival when treated with doxorubicin.

Androstadienes↗
PubMed1999

Expression of the wild-type insulin-like growth factor II receptor gene suppresses growth and causes death in colorectal carcinoma cells.

The insulin-like growth factor II receptor (IGFIIR) has been implicated as a tumor suppressor gene in human malignancy. Frequent mutation, loss of heterozygosity, and microsatellite instability (MSI) directly affecting the IGFIIR gene have been reported in several primary human tumor types. However, to our knowledge, dynamic functional evidence of a growth-suppressive role for IGFIIR has not yet been provided. We identified one MSI-positive colorectal carcinoma cell line, SW48, with monoallelic mutation in IGFIIR identical to that seen in primary colorectal carcinomas. A zinc-inducible construct containing the wild-type IGFIIR cDNA was stably transfected into SW48 cells. Growth rate and apoptosis were compared between zinc-treated, untreated, and untransfected cells. A twofold increase in IGFIIR protein expression was detected after zinc treatment in discrete clonal isolates of transfected SW48 cells. Moreover, zinc induction of exogenous wild-type IGFIIR expression reproducibly decreased growth rate and increased apoptosis. These data prove that wild-type IGFIIR functions as a growth suppressor gene in colorectal cancer cells and provide dynamic in vitro functional support for the hypothesis that IGFIIR is a human growth suppressor gene.

Adenocarcinoma↗
PubMed1999

HIV seroprevalence & awareness about AIDS among pregnant women in rural areas of Pune district, Maharashtra, India.

The present unlinked anonymous study was done among sexually active rural women to assess the extent of spread of HIV and its awareness. Peripheral blood samples were collected on filter paper strips from 1251 pregnant women residing in villages in three Primary Health Centres in Pune district of Maharashtra. Elutes were tested for HIV antibodies in two different ELISA systems. Awareness on HIV/AIDS was assessed using a structured questionnaire. Fifteen (1.2%) samples were detected to be HIV seropositive. HIV seroprevalence was significantly higher among villages situated close to highways (P < 0.025). Majority (> 95%) of the participating women were housewives. Although 70 per cent were aware of the existence of AIDS, only 33 per cent knew about all the main modes of HIV transmission. Their main sources of information on AIDS were health camps, health workers (70%) and television (45%). Awareness was associated with higher level of literacy (P < 0.001). Many women had misconceptions about the modes of spread of HIV. Greater emphasis needs to be placed on instituting long-term and sustainable strategies to create awareness among young couples with an emphasis on involvement of health workers in rural areas.

Acquired Immunodeficiency Syndrome↗
PubMed1994

Osmolyte mediation of T7 DNA polymerase and plasmid DNA stability.

The thermal stability of T7 DNA polymerase and pGEM4Z plasmid DNA in the presence and absence of the osmolyte N-methylglycine (sarcosine) was determined by means of UV spectroscopy. The decrease in melting temperature observed upon addition of sarcosine to solutions containing the plasmid DNA is linear with the concentration of sarcosine present. The enthalpy of the transition is also linear in its relationship to the melting temperature, and the entropy of the transition is linear in the natural log of the melting temperature. The slopes of both the entropic and enthalpic plots are equal. Destabilization of the plasmid DNA is observed to be entropically driven. The melting temperature of the T7 DNA polymerase complex is increased from 41 degrees C by addition of sarcosine to the solution. The relationship between the amount of sarcosine added and the melting temperature is linear, with a temperature of 61 degrees C observed for a 6 M solution. No clear trend of the effect of sarcosine on the enthalpy or entropy of the transitions could be observed.

DNA↗
PubMed1994

Oral fluconazole therapy for keratomycosis.

A 12-year-old girl with fungal keratitis following injury with a vegetable foreign body was successfully treated with oral fluconazole, a new antifungal agent. The infection resolved completely with good visual recovery. Fluconazole was well tolerated systemically.

Administration, Oral↗
PubMed1993

Postirradiation exposure to hypotonic saline shows normal damage processing in radiation-sensitive cell lines.

Three pairs of cell lines (one human and two Chinese hamster ovary (CHO) cell lines) each comprising a cell line with a normal radiation response and a radiation-sensitive mutant, were evaluated for recovery of potentially lethal damage (PLDR) and recovery of sublethal damage (SLDR). In all cases, the normal cell lines (GM1522, human; AA8-4 and K1, CHO) exhibited capacity for PLDR and SLDR was also normal in the two CHO lines. For the mutants (GM3395, human AT; V3 and 5-11, CHO) there was no ability for PLDR and SLDR was also absent in the two CHO cell lines. Postirradiation exposure to hypotonic NaCl solutions resulted in fixation of radiation damage in all the cell lines. This form of damage is repaired if left unperturbed after irradiation. This shows that cells have a large capacity for repair of this form of damage which accounts for much greater changes in survival than those observed in conventional PLDR experiments. These data show that the mutant cell lines retained their capacity to repair the damage which was susceptible to postirradiation fixation by anisotonic salt solutions. In addition, initial (i.e. preirradiation) DNA polymerase activities were measured in the four CHO cell lines; they were not correlated to radiation sensitivity.

Animals↗
PubMed1992

Serum beta 2 microglobulin levels in HIV seropositive persons.

As beta 2 microglobulin (B2M) has been found to be elevated in immunological disorders and also in HIV infection, its levels were studied in 475 HIV seropositive, asymptomatic persons; 101 HIV seronegative persons from high risk groups for HIV and 99 healthy controls. The B2M levels in asymptomatic HIV seropositives are found to be significantly higher than healthy controls (1.0 mg/1 to 2.7 mg/1, P less than 0.001) and HIV seronegatives from high risk groups (1.1 mg/1 to 2.7 mg/1, P less than 0.001). Two hundred and thirty four (49.26%) seropositives showed increased levels of serum B2M. Thus, quantitative analysis of B2M may be useful as an early nonspecific marker of HIV infection and immune dysfunction. The prognostic value of B2M was assessed in a follow up study of 54 HIV seropositives in a 2 yr period. Within this period, B2M levels were found to be significantly increased in these subjects (1.2 mg/1 to 4.6 mg/1, P less than 0.001). Three of the subjects who showed high increase in the B2M levels, progressed to AIDS-related condition, whereas one progressed to persistent generalised lymphadenopathy. Thus, the rising levels of B2M appears to correlate well with disease progression.

Follow-Up Studies↗
PubMed1992

HIV-2 antibodies in serum samples from Maharashtra state.

Two hundred serum samples were tested to detect the presence of HIV-2 infection in Maharashtra state. The serum samples were derived from various groups including those with high risk behaviour for HIV infection. All samples were tested by three combined HIV-1 and HIV-2 ELISA kits. The reactivity was confirmed by LiaTek HIV 1+2 immunoblot. The study confirmed that HIV-2 infection exists in Maharashtra, as 14 samples showed antibodies to HIV-2 and 14 showed antibodies to both HIV-1 and HIV-2. Antibodies to HIV-2 or HIV-1 and HIV-2 were detected mainly in persons with high risk behaviour.

Acquired Immunodeficiency Syndrome↗
PubMed1991

Circulating immune complexes in healthy, HIV-antibody positive subjects.

The components of the circulating immune complexes (CICs) were characterised in asymptomatic HIV seropositive individuals. Forty four of 214 individuals (20.56%) showed the presence of CICs. Specific HIV anti-HIV CICs were detected in 33 of them (75%). The isotypic specificity of antibodies found to these CICs was measured. IgG and IgA immunoglobulin classes were detected in CICs.

Antigen-Antibody Complex↗
PubMedDate not supplied

Thermal radiosensitization in two pairs of CHO wild-type and radiation-sensitive mutant cell lines.

Two pairs of Chinese hamster ovary cell lines, each comprising a parental normal-responding and a radiation-sensitive mutant, were tested for their X-radiation sensitivity, hyperthermia sensitivity and thermal radiosensitization. The relative radiation responses were similar to those already published in the literature, indicating that the mutant cell lines, V-3 and 5-11, were extremely sensitive to radiation while their parental strains (AA8-4 and K1) exhibited normal sensitivity. The hyperthermia response of V3 was the same as that of the parental cell line AA8-4; the thermal sensitivity of 5-11 was greater than that of its parental line K1 but this difference decreased at higher temperatures (greater than 43 degrees C). These data indicate that the hyperthermia response and its targets are probably not related to the mutation-causing radiation sensitivity in the two pairs of parent-mutant cell lines. When heat and radiation were combined, the thermal enhancement of radiation sensitivity for different heat treatments was the same or larger in the mutant than in the parental cell lines. The mutants are known to be deficient in potentially lethal damage repair (PLDR), sublethal damage repair (SLDR) and repair of double-stranded DNA breaks; however, these deficiencies did not impair thermal radiosensitization, indicating that thermal radiosensitization may not be through these pathways in these particular mutants. In addition, the parental cell lines exhibited a significant sequence dependence of thermal radiosensitization (i.e. whether heat was applied before or after irradiation) at low survival levels (10(-4); this sequence dependence was very small to absent in the mutant cell lines at these survival levels.

Animals↗
PubMedDate not supplied

Human immunodeficiency virus-1 infection in spouses of seropositive individuals.

BACKGROUND: Unprotected sex can lead to transmission of the human immunodeficiency virus-1 (HIV-1) to the spouse of an infected individual. We studied the incidence of HIV-1 infection in the spouses of cases diagnosed to have HIV-1 infection by serology and the polymerase chain reaction (PCR). METHODS: Blood samples collected from 9 index cases and their respective spouses were tested for HIV-1 infection by ELISA, Western blot (WB) and PCR as well as from 10 healthy individuals with no high-risk behaviour. DNA extracted from both plasma and peripheral blood mononuclear cells was amplified by PCR, using multiple primer pairs for distinct regions of the HIV-1 genome. Specificity of the PCR product was demonstrated by hybridization to an oligonucleotide probe. RESULTS: All the index cases which were seropositive by ELISA and WB were also positive by PCR of plasma extracted DNA. Eight of the spouses were seronegative. Of these seven were positive by PCR--one spouse was negative by ELISA but showed a p55 band on WB and was positive by PCR. One spouse was negative by serology and PCR. The spouse belonging to the lone concordant couple was positive by serology and PCR. Except for one index case, PCR signals were obtainable only from DNA extracted from plasma but not from the DNA extracted from peripheral blood mononuclear cells. The control samples were negative by serology and PCR. CONCLUSION: It is possible to detect HIV-1 infection by PCR using DNA extracted from plasma even when the individuals are negative by ELISA and WB. It can help in the early counselling of HIV infected persons and their spouses.

AIDS Serodiagnosis↗