PubMed Health⌕ Search

Biomedical subjects

M Saini

Publications and source records attributed to M Saini.

At least 19 recordsLinked to original sources
PubMed2006

Identification of a 66 kDa Haemonchus contortus excretory/secretory antigen that inhibits host monocytes.

A 66 kDa adult Haemonchus contortus excretory/secretory (E/S) antigen was identified in Western blot by reaction with sera from the infected goats. The protein was purified from the adult worm extract and E/S products by anion exchange and ConA-Sepharose chromatography. The purified protein inhibited monocyte function in vitro as judged by decreased production of hydrogen peroxide and nitric oxide in the culture medium. The protein also caused proliferation of peripheral blood mononuclear cells. The absence of protein in the free living L3 larvae suggests that the expression of this protein coincides with the adaptation to the parasitic life. A correlation of antibody titre and worm burden was observed in the infected goats with higher antibody levels in high worm burdened animals. Anti-protein antibody caused loss of adult worm motility in vitro resulting in the death of the parasite. The fact that the protein is recognized by the host together with in vitro killing of adult parasites by antibodies makes this protein a promising candidate for vaccination trial.

Abomasum↗
PubMed2006

A DNA vaccine that encodes rabies virus glycoprotein lacking transmembrane domain enhances antibody response but not protection.

Rabies virus (RV) glycoprotein (gp) consists of three domains: cytoplasmic, transmembrane and ectodomain. It occurs in a complete, membrane-bound form within the infected cell, but it is released from them in a deleted, secreted form lacking the transmembrane domain. This study was performed to test the importance of the transmembrane domain for the capability of the RV gp gene, introduced into mice via a recombinant plasmid (DNA vaccine), to induce immune response and protection against challenge. Although the antibody response to the secreted form of gp was higher than that to complete gp, the protective efficacy of the respective DNA vaccine against challenge was not better than that of the DNA vaccine inducing complete gp. This indicates that the transmembrane domain of RV gp is important for generating protection against rabies and should be present in RV DNA vaccines.

Animals↗
PubMed2005

Fasciola gigantica cathepsin-L cysteine proteinase in the detection of early experimental fasciolosis in ruminants.

Cathepsin-L cysteine proteinase was purified from Fasciola gigantica regurgitant by two-step alcoholic fractionation, followed by ion-exchange chromatography. The purification strategy was evolved to eliminate other contaminating proteins co-precipitating with the purified proteinase during alcoholic fractionation. The enzyme was stable on long-term storage at -20 degrees C rendering it more suitable for field diagnostic use. The purified cathepsin-L cysteine proteinase was assayed for detection of F. gigantica experimental infection in sheep and buffaloes and could detect infection, as early as 4 weeks post-infection by ELISA, Western blotting and Dipstick ELISA. The 28-kDa cathepsin-L cysteine proteinase seems a promising antigen for the diagnosis of tropical fasciolosis in domestic animals.

Alcohols↗
PubMed2005

Immunogenic and antigenic properties of recombinant soluble glycoprotein of rabies virus.

Rabies virus glycoprotein is a type I transmembrane protein exposed on the surface on the mature virus particle that induces virus neutralizing antibodies. In the present study, 60 amino acid C-terminal hydrophobic anchor (transmembrane) and cytoplasmic domains of glycoprotein were deleted from full-length glycoprotein and fused with polyhistidine tag. The N-terminal viral signal peptide was also replaced with CD33 signal peptide for efficient secretion in mammalian cells. Following transfection of Madin Darby bovine kidney (MDBK) cells with plasmid encoding this soluble form of glycoprotein, polyclonal populations of stably transfected resistant cells were obtained after G418 selection. The protein was expressed as a glycosylated protein and secreted outside the cells utilizing N-terminal CD33 signal peptide. The secreted soluble glycoprotein was purified from cell culture supernatant by Ni--agarose affinity chromatography utilizing C-terminal polyhistidine tag. Like full-length glycoprotein, the expressed recombinant soluble glycoprotein was found to be immunogenic when injected in rabbits. In this study, we have assessed the potential of recombinant soluble glycoprotein as diagnostic antigen in ELISA and found that this recombinant protein can be used as diagnostic antigen in ELISA for detecting anti-glycoprotein antibodies in immunized host.

Animals↗
PubMed2005

Surgical removal of primary central nervous system lymphomas (PCNSL) presenting as space occupying lesions: a series of 33 cases.

AIMS: In this study we present a series of 33 patients with primary CNS lymphomas (PCNSL), many presenting with acute signs of increased intracranial pressure due to large space occupying lesions. METHODS: A series of 32 PCNSL patients for a total of 33 tumours treated from 1986 to 2000 in the Neurosurgical Department were reviewed. RESULTS: Radiotherapy and chemotherapy improved survival. No benefit could be demonstrated for the role of surgery. CONCLUSIONS: Our data confirm previous reports about the role of radiation and chemotherapy in the treatment of PCNSL's. Surgery might have a role in a selected subset of patients presenting with large single space occupying lesions and deteriorating neurological status.

Adult↗
PubMed2004

A model for intratumoural chemotherapy in the rat brain.

To achieve the best reproducibility in rat brain tumour models several injection techniques have been used. Although stereotactic cell injections have proved to be effective and reliable, they are expensive and time consuming. A new permanently implanted device is presented here. It allows precise cell delivery for best tumour reproducibility, and it can be left in place for future injections at the exact same location, such as intratumoural chemotherapy. A Teflon tube was mounted on a disc, inserted into the rat brain and sealed to the skull. The device was tested in two rat strains (Wistar and New Zealand Nude rats) with two different glioma cell lines (9L and C6). Rats were treated with placebo to determine if repeated treatments had an effect on the device placement, or if device-related morbidity was induced. Analysis of brain sections showed that the device path was always within the tumour. The device never moved or came off the scalp. Both Wistar rats and NZ nude rats tolerated the device well. No morbidity or mortality was observed, regardless of the presence of the device; no infections were seen. Biocompatible, non-irritating and well tolerated, such a device can be used for reproducible tumour cell injections and repeated intralesional delivery of drugs.

Animals↗
PubMed2004

High nucleotide and amino acid sequence similarities in tumour necrosis factor-alpha amongst Indian buffalo (Bubalus bubalis), Indian cattle (Bos indicus) and other ruminants.

Tumour necrosis factor-alpha (TNF-alpha) mRNA from Indian water buffalo (Bubalus bubalis) and Indian cattle (Bos indicus) was reverse transcribed and amplified using reverse transcriptase-polymerase chain reaction (RT-PCR). The nucleotide sequences of cDNAs were determined after cloning into pGEM-T-Easy vector (Promega, Madison, WI) and compared with reported nucleotide sequences of TNF-alpha cDNA from other species. The nucleotide sequences of TNF-alpha from Indian cattle revealed significantly high similarities at nucleotide (99.2%) and amino acid (100%) levels with those of cattle (Bos taurus; Zebu). The sequences from buffalo had 98.4% nucleotide and 99.1% amino acid similarities with Indian cattle, indicating functional cross-reactivity. One amino acid deletion at position 63 and one substitution (A-->P) at position 64 were observed in buffalo compared with Indian cattle. The amino acid deletion at position 63 was predicted due to differences in pre-mRNA splicing.

Amino Acid Sequence↗
PubMed2002

A study of prevalence of health problems in asymptomatic elderly individuals in Delhi.

A survey was conducted in asymptomatic aged individuals (> or = 60 years) in The National Capital Territory of Delhi for the prevalence of major health problems like hypertension, diabetes mellitus and respiratory diseases. A total of 200 individuals (100 males and 100 females) were studied over a period of three months in 1998-99. Hypertension was defined as BP > or = 140/90 mmHg (JNC VI criteria), while diabetes mellitus was diagnosed if fasting whole blood sugar was 120 mg/dl or more (WHO criteria). Diagnosis of other health problems was based on relevant history and physical examination. Prevalence of hypertension in the study group was 32.5 per cent (more in males). Of these 18 per cent and 4.2 per cent had isolated systolic and diastolic hypertension, respectively. Prevalence of diabetes mellitus in the same population was 13.0 per cent. Both diseases were more prevalent in urban population. A high prevalence of respiratory disorders was observed (pulmonary tuberculosis 16 per cent, COPD 10 per cent, asthma 4.5 per cent). Cataract was present in 7.5 per cent while 1.5 per cent had symptoms of urinary tract infection. History of Jaundice was present in 3.5 per cent. Three per cent each had a history suggestive of IHD and TIA, respectively. Proteinuria and glycosuria was seen in 22.2 and 7.6 percent, respectively. A large percentage of the study group (34.4 per cent) had asymptomatic ECG abnormalities.

Aged↗
PubMed2001

Transfection of mGSTA4 in HL-60 cells protects against 4-hydroxynonenal-induced apoptosis by inhibiting JNK-mediated signaling.

The mammalian alpha-class glutathione S-transferase (GST) isozymes mGSTA4-4, rGSTA4-4, and hGSTA4-4 are known to utilize 4-hydroxynonenal (4HNE) as a preferred substrate. During the present studies, we have examined the effect of transfecting human myeloid HL-60 cells with mGSTA4, on 4-HNE-induced apoptosis and the associated signaling mechanisms. Results of these studies show that treatment of the wild-type or vector-only-transfected HL-60 cells with 20 microM 4-HNE caused apoptosis within 2 h. The cells transfected with mGSTA4 did not undergo apoptosis under these conditions even after 4 h. In the wild-type and vector-transfected cells, apoptosis was preceded by JNK activation and c-Jun phosphorylation within 30 min, and an increase in AP-1 binding within 2 h of treatment with 20 microM 4-HNE. In mGSTA4-transfected cells, JNK activation and c-Jun phosphorylation were observed after 1 h, and increased AP-1 binding was observed after 8 h under these conditions. In the control cells, 20 microM 4-HNE caused caspase 3 activation and poly(ADP-ribose) polymerase cleavage within 2 h, while in mGSTA4-transfected cells, a lesser degree of these effects was observed even after 8 h. Transfection with mGSTA4 also provided protection to the cells from 4-HNE and doxorubicin cytotoxicity (1.6- and 2.6-fold, respectively). These results show that 4-HNE mediates apoptosis through its effects on JNK and caspase 3, and that 4-HNE metabolizing GST isozyme(s) may be important in the regulation of this pathway of oxidative-stress-induced apoptosis.

Aldehydes↗
PubMed2001

Role of glutathione S-transferases in protection against lipid peroxidation. Overexpression of hGSTA2-2 in K562 cells protects against hydrogen peroxide-induced apoptosis and inhibits JNK and caspase 3 activation.

The physiological significance of the selenium-independent glutathione peroxidase (GPx) activity of glutathione S-transferases (GSTs), associated with the major Alpha class isoenzymes hGSTA1-1 and hGSTA2-2, is not known. In the present studies we demonstrate that these isoenzymes show high GPx activity toward phospholipid hydroperoxides (PL-OOH) and they can catalyze GSH-dependent reduction of PL-OOH in situ in biological membranes. A major portion of GPx activity of human liver and testis toward phosphatidylcholine hydroperoxide (PC-OOH) is contributed by the Alpha class GSTs. Overexpression of hGSTA2-2 in K562 cells attenuates lipid peroxidation under normal conditions as well as during the oxidative stress and confers about 1.5-fold resistance to these cells from H(2)O(2) cytotoxicity. Treatment with 30 microm H(2)O(2) for 48 h or 40 microm PC-OOH for 8 h causes apoptosis in control cells, whereas hGSTA2-2-overexpressing cells are protected from apoptosis under these conditions. In control cells, H(2)O(2) treatment causes an early (within 2 h), robust, and persistent (at least 24 h) activation of JNK, whereas in hGSTA2-2-overexpressing cells, only a slight activation of JNK activity is observed at 6 h which declines to basal levels within 24 h. Caspase 3-mediated poly(ADP-ribose) polymerase cleavage is also inhibited in cells overexpressing hGSTA2-2. hGSTA2 transfection does not affect the function of antioxidant enzymes including GPx activity toward H(2)O(2) suggesting that the Alpha class GSTs play an important role in regulation of the intracellular concentrations of the lipid peroxidation products that may be involved in the signaling mechanisms of apoptosis.

Apoptosis↗
PubMed2001

Induction of immune responses in cattle with a DNA vaccine encoding glycoprotein C of bovine herpesvirus-1.

A DNA vaccine expressing glycoprotein C (gC) of bovine herpesvirus-1 (BHV-1) was evaluated for inducing immunity in bovines. The plasmid encoding gC of BHV-1 was injected six times intramuscularly or intradermally into calves at monthly intervals. After immunization by both routes neutralizing antibody and lymphoproliferative responses developed. The responses in the intradermally immunized calves were better than those in calves immunized intramuscularly. However, the intradermal (i.d.) route was found to be less efficacious when protection against BHV-1 challenge was compared. Following intranasal BHV-1 challenge, all immunized calves demonstrated a rise in IgG antibody titre on day 3, indicating an anamnestic response. The control non-immunized calf developed a neutralizing antibody response on day 7 post-challenge. The immunized calves showed a slight rise in temperature and mild clinical symptoms after challenge. The intramuscularly immunized calves showed earlier clearance of challenge virus compared with intradermally immunized calves. These results indicate that DNA immunization with gC could induce neutralizing antibody and lymphoproliferative responses with BHV-1 responsive memory B cells in bovines. However, the immunity developed was not sufficient to protect calves completely from BHV-1 challenge.

Animals↗
PubMed2001

Characterization of toxin from cheilanthes fern and its effect on lymphocyte proliferation and DNA fragmentation.

Thin layer chromatography of aqueous extract of whole Cheilanthesfarinosa fern indicated the presence of ptaquiloside or ptaquiloside like compound, coinciding Rf values with that of Pterosin B standard. HPLC analysis revealed the presence of 26.3 mg/kg ptaquiloside. In vitro studies of the aqueous extract on lymphocyte culture revealed a correlation between stimulative indices and concentration of aqueous extract. Stimulation in lymphocyte proliferation was in order of bracken > cheilanthes > ConA> ptaquiloside standard. On incubation of lymphocyte with aqueous extract of ferns, no DNA damage was observed in isolated DNA.

Animals↗
PubMed1999

Effects of mGST A4 transfection on 4-hydroxynonenal-mediated apoptosis and differentiation of K562 human erythroleukemia cells.

Cellular levels of downstream products of membrane lipid oxidation appear to regulate differentiation in K562 human erythroleukemia cells. 4-Hydroxynonenal (4-HNE) is a diffusible and relatively stable product of peroxidation of arachidonic and linoleic acids, cellular levels of which are regulated through metabolism to glutathione (GSH) conjugate by glutathione S-transferases (GSTs). A group of immunologically related alpha-class mammalian GSTs expressed in mice (mGST A4-4), rat (rGST A4-4), human (hGST A5.8), and other species, as well as the more distantly related human hGST A4-4, preferentially utilize 4-HNE as a substrate and are suggested to be major determinants of intracellular levels of 4-HNE. Present studies were designed to examine the effects of 4-HNE on K562 cells and to study the effect of transfection of mGSTA4-4 in these cells. Exposure of K562 cells to 20 microM 4-HNE for 2 h resulted in a rapid erythroid differentiation of K562 cells, as well as apoptosis evidenced by characteristic DNA laddering. Stable transfection of cells with mGST A4-4 resulted in a fivefold increase in GST-specific activity toward 4-HNE compared with wild-type or vector-only transfected cells. The mGST A4-4-transfected cells were resistant to the cytotoxic, apoptotic, and differentiating effects of 4-HNE. The mGST A4 transfection also conferred resistance to direct oxidative stress (IC(50) of H(2)O(2) 22, 23, and 35 microM for wild-type, vector-transfected, and mGST A4-transfected cells, respectively). mGST A4-4-transfected cells also showed a higher rate of proliferation compared with wild-type or vector-transfected K562 cells (doubling time 22.1 +/- 0.7, 31 +/- 1.2, and 29 +/- 0.6 h, respectively). Cellular 4-HNE levels determined by mass spectrometry were lower in mGST A4-4-transfected cells compared to cells transfected with vector alone (5.9 pmol/5 x 10(7) cells and 62.9 pmol/5 x 10(7) cells, respectively). Our studies show that 4-HNE can induce erythroid differentiation in K562 cells and that overexpression of mGST A4 suppresses 4-HNE levels and inhibits erythroid differentiation and apoptosis.

Aldehydes↗
PubMed1999

Morphometrical characterization of two glioma models in the brain of immunocompetent and immunodeficient rats.

Although several glioma models exist, systematic morphometrical studies on such experimental tumors are lacking. The purpose of this study was the quantitative assessment of how rat strains, cell lines, injection techniques and location affect tumors reproducibility and histopathological features. Glioma cells were implanted in 3 brain locations, with different injection techniques (free hand, stereotactic, water-tight device), variable volumes, cell concentrations and infusion rates. Tumors were developed from 2 rat glioma cell lines (9L and C6) in immunocompetent (Wistar and Fischer 344) and immunodeficient rats (New Zealand). Animals underwent daily neurological examination. At the scheduled time the tumors were macro and microscopically evaluated and a quantitative morphometrical analysis was performed. C6 gliomas appeared very infiltrative and irregularly shaped; 9L gliomas showed, by using the same injection technique, a grossly regular shape. Margins at the tumor-brain interface were macroscopically demarcated in the immunocompetent rats. In the nude rats, 9L tumors appeared microscopically more infiltrative, although regularly shaped, with a closer morphological resemblance to human gliomas. The implantation in the frontal area, anterior to the nucleus caudatus (3 mm anterior the coronal suture) gave reproducible tumor shape and size, no hydrocephalus and no early neurological deterioration. The use of a stereotactic technique or of a water-tight device, small volume (< 10 microl) of cell suspension, low infusion rate were useful to reduce morbidity and to improve data reproducibility. No difference in morbidity and mortality were observed in immunocompetent and immunodeficient rats. The 9L glioma model with stereotactic implantation constitutes a good option for reliable morphometrical evaluation of tumor growth. We propose a location for tumor implantation anterior to the nucleus caudatus. This produced the longest symptom-free survival.

Animals↗
PubMed1999

A new xenograft model of primary central nervous system lymphoma.

The management of primary lymphoma of the central nervous system (PCNSL) remains controversial and patients' outcome dismal. In order to investigate new selective therapeutic strategies in a controlled system, a reproducible model of PCNSL in nude rats was developed and characterized. Human B lymphoma cells (BL2) were implanted in the brain frontal area in New Zealand nude rats through a silastic device sealed to the skull. Fifteen and 30 days post-implantation, animals were sacrificed. An autopsy was performed. Representative brain sections were cut and examined for the presence of lymphoma. Immunohistochemistry was performed for proliferation (MIB1-Ki67), a B-cell marker (L26-CD20), a T-cell marker (UCHL1-CD45RO). The analysis of the brains showed tumor growth in 88% of the rats. No mortality was observed. At autopsy no extracerebral, spinal or cerebellar metastasis were found. Microscopically the brain tumors appeared non-encapsulated, highly vascularized, with a characteristic perivascular and diffuse lymphomatous spread in the parenchyma. Immunohistochemistry showed a marked positivity of the tumor cells for L26. Tumor cells were negative for UCHL1. Mean proliferation rate was 30%. The device was well tolerated and caused no local infection. Controlled studies on PCNSL in animal models are lacking. This PCNSL model in nude rats reproduces the histology and location of human CNS lymphoma. Tumor dimensions are within the resolution limits of CT and MRI and therefore suitable for stereotactic therapy. This model provides a tool to test new chemo and radiotherapeutical strategies in a controlled fashion.

Animals↗