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T Das

Publications and source records attributed to T Das.

At least 19 recordsLinked to original sources
PubMed1996

Inhibition of human parainfluenza virus-3 replication by interferon and human MxA.

We have investigated the IFN-mediated inhibition of human parainfluenza virus-3 (HPIV-3) replication in cultured human A549 cells. IFN-alpha inhibited the virus yield significantly with concomitant reduction of viral RNA accumulation by more than 90%. Further studies indicated that the inhibitory action of IFN was at the level of primary transcription of HPIV3 replication. Since the IFN-inducible protein, MxA, has been shown to inhibit virus replication in several RNA viruses, we examined the role of MxA in HPIV-3 replication using a stably transfected human glioblastoma cell line expressing MxA. In these cells HPIV-3 replication was decreased by more than 100-fold depending on the virus dosage used with concomitant inhibition of viral RNA synthesis by about 80%. However, the viral primary transcription was not affected in this MxA-producing cell line. In contrast, in the parental cell line IFN-mediated inhibition occurred at the primary transcription step of HPIV-3 replication. These data suggest that in addition to MxA, other IFN-inducible proteins are involved in the anti-HPIV-3 effect of IFN in both the cell lines used.

Antiviral Agents↗
PubMed1996

Expression of the human MxA protein is associated with hyperphosphorylation of VSV P protein in human neural Cells.

Constitutive expression of the type I interferon-inducible human cytoplasmic MxA protein has been shown to interfere with primary transcription of vesicular stomatitis virus (VSV) in tissue culture cells. As phosphorylation of the VSV P protein has been linked to its ability to stimulate viral transcription, we analyzed the phosphorylation status of this protein in human brain cells (U-87) stably transfected with MxA. We observed a general increase in cellular kinase activity in the presence of MxA, affecting both cellular proteins and VSV P protein. Phosphorylation of the latter was up to threefold higher both in vivo and in vitro. In vitro phosphorylation of recombinant VSV P protein could be enhanced in MxA-negative cell extracts after exogenous addition of recombinant His-MxA. Biochemical evidence and phosphorylation of a mutant P protein lacking the recognized casein kinase II (CKII) sites suggested that hyperphosphorylation of VSV P protein was not due to a stimulation of CKII. We thus propose that expression of MxA in human brain cells is associated with the stimulation of a cellular kinase that is active in phosphorylating both cellular target proteins and VSV P protein.

Casein Kinase II↗
PubMed1996

Dietary garlic extract in modifying clastogenic effects of inorganic arsenic in mice: two-generation studies.

Mice are fed by gavage crude garlic extract (100 mg/kg b.wt.) for 30 consecutive days. One set was administered sodium arsenite (0.1 mg/kg b.wt.) simultaneously. Another set was treated with sodium arsenite only. Mice given distilled water were kept as negative control. Exposed mice from each set were sacrificed and bone marrow preparations examined for chromosomal aberrations and damaged cells. Sodium arsenite is a strong clastogen and the effects were reduced to a significant level by prolonged administration of garlic extract. For F1 studies, exposed male mice were mated with exposed female mice, and the progeny examined. In the progeny, clastogenic effects of sodium arsenite persisted in a lower degree, indicating that the metal is able to cross the transplacental barrier. There was no statistically significant difference between the effect in progeny of parents only given sodium arsenite when given simultaneously for prolonged periods in the parents; however, the effect is meagre in the next generation.

Animals↗
PubMed1996

A 50 KDa protein modulates guanine nucleotide binding of transglutaminase II.

Regulation of cellular response is an important mechanism for controlling cellular functions. The transmembrane signaling of the hormone receptors is regulated by GTP-binding proteins (GTPases) and their associated proteins. Our previous studies demonstrated that the bifunctional GTP-binding protein, G alpha h (transglutaminase II), consistently copurified with an approximately 50 kDa protein (G Beta h) which is dissociated from G alpha h upon activation with GTP gamma S or AlF4-. Present immunological and biochemical studies on the regulation of the GTPase cycle of G alpha h, which involves the alpha 1-adrenoceptor and 50 KDa G beta h, reveal that the 50 kDa protein is indeed a G alpha h-associated protein and down regulates functions of G alpha h. Thus, polyclonal antibody against G Beta h coimmunoprecipitates GDP-bound G alpha h but not the GDP-AlF4--bound form. The GTP gamma S binding and GTPase activity of G alpha h are inhibited in a G beta h concentration dependent manner. Supporting this notion, G beta h accelerated GTP gamma S release from G alpha h and changes the affinity of G alpha h from GTP to GDP. Moreover, the ternary complex preparation exhibits TGase activity that is inhibited in the presence of the alpha 1-agonist and GTP. The GTP gamma S binding by the ternary complex, consisting of the alpha 1-agonist, the receptor, and Gh, is also inhibited by G beta h. The inhibition of GTP gamma S binding with the ternary complex requires a > or = 2.7-fold higher concentration of G beta h than the G alpha h alone, indicating that the receptor enhances the affinity of G alpha h for GTP. In addition, G beta h copurifies with an alpha 1-agonist, adrenoceptor, and G alpha h ternary complex, showing that the complex is a heterotetramer. Our data also suggest that G beta h does not directly interact with alpha 1-adrenoceptor. These findings clearly demonstrate that G alpha h associates with a novel protein which modulates the affinity of G alpha h for guanine nucleotides and that the GDP-bound Gh is the ground state for the counterpart activator, the alpha 1-adrenoceptor, in this signaling system.

Animals↗
PubMed1996

Effects of crude garlic extract on mouse chromosomes in vivo.

Three concentrations (25, 50 and 100 mg/kg body weight) of fresh garlic (Allium sativum L.) were administered daily by gavage to Swiss albino mice for different durations up to 60 days. These concentrations had been observed to protect significantly against effects of known clastogens. The endpoints scored were frequencies of chromosomal aberrations and damaged cells induced in bone marrow preparations. These parameters were found to be directly dose dependent and after an initial enhancement at 7 days, were reduced following prolonged exposure for 30 and 60 days to the low level observed at 24 hr. Therefore, administration of a low concentration of garlic extract daily is suggested for at least 30 days to obtain the maximum benefit of the extract in protecting against the clastogenic effects of known genotoxicants.

Administration, Oral↗
PubMed1996

Presumed noninfectious endophthalmitis after cataract surgery.

PURPOSE: To study the clinical features and results of conservative management in cases of presumed noninfectious endophthalmitis after cataract surgery. SETTING: L.V. Prasad Eye Institute, Hyderabad, India. METHODS: This retrospective study evaluated cases of presumed noninfectious endophthalmitis occurring after cataract surgery in a hospital-based population. Patients with minimal hypopyon and decreased visual acuity unassociated with pain, lid edema, conjunctival chemosis, corneal abscess, and obvious vitreous involvement were treated conservatively. The mainstays of treatment were topical and systemic steroids. Four eyes with clinical evidence of worsening required surgical intervention. Main outcome measures were resolution of inflammation and final visual acuity. RESULTS: In 23 of 27 patients, the inflammation resolved with medical therapy alone; 21 had a final visual acuity of 20/50 or better. In 3 of the 4 eyes that had surgery, the culture was positive for low virulent organisms; all had a satisfactory outcome. One eye with preoperative glaucoma developed absolute glaucoma with resolution of inflammation. CONCLUSION: Noninfectious endophthalmitis invariably presents with hypopyon and decreased vision unassociated with pain and lid edema. A trial of medical therapy with corticosteroids as the mainstay of treatment is often rewarding. Frequent monitoring and preparation for surgical intervention if required are key.

Adolescent↗
PubMed1996

Sensitivity and predictability of vitreous cytology, biopsy, and membrane filter culture in endophthalmitis.

PURPOSE: Forty-seven consecutive patients with endophthalmitis were prospectively studied to: 1) compare the undiluted vitreous biopsy culture and membrane filter culture of vitrectomy cassette fluid; 2) determine the sensitivity and specificity of initial smear examinations in relation to final culture; and 3) characterize vitreous cytology in culture positive and negative endophthalmitis. METHODS: Examinations included smears (Gram stains, Diff Quik [Bacto Laboratories Pvt. Ltd., Liverpool, Australia] and unstained) and aerobic/anaerobic culture of undiluted vitreous biopsy specimens and diluted vitreous collected in the cassette. The cassette fluid was passed through 5-microns polycarbonate filter for cytology and a 0.22-microns polyvinylidene difluoride millipore filter for culture. RESULTS: By either method culture results were positive in 27 (57.4%) patients. Culture positive of the vitreous biopsy alone was 44.6% and cassette fluid alone was 49% (P > 0.05). In the initial smear examination the sensitivity and specificity of the Gram stain were 66.6% and 84.2%, respectively. comparatively, Diff Quik provided lower sensitivity and specificity at 40.7% and 80.9% respectively. Examination of wet film of vitreous samples did not provide any useful information. Although polymorphs were seen in large numbers in all cultures that yielded positive results on examination, they also were seen in 80% of the sterile samples. There was no significant difference in the quantity of macrophages observed in infected and noninfected samples. CONCLUSIONS: Initial smear examination and cytology have limited roles in the diagnosis of infectious endophthalmitis. Obtaining cultures of both an undiluted vitreous biopsy sample and the vitrectomy cassette fluid has a significant advantage compared with culture of only one sample.

Adolescent↗
PubMed1996

Expression of L protein of vesicular stomatitis virus Indiana serotype from recombinant baculovirus in insect cells: requirement of a host factor(s) for its biological activity in vitro.

The 241-kDa large (L) protein of vesicular stomatitis virus (VSV) Indiana serotype, a multifunctional catalytic subunit of the viral RNA polymerase, has been expressed in Spodoptera frugiperda cells infected with recombinant baculovirus BacPAK6-L containing the L gene under the control of a polyhedrin promoter. The recombinant L protein was biologically active and supported viral mRNA synthesis in vitro. When the expressed L protein was purified by phosphocellulose column chromatography, it eluted in two peaks, one at 0.4 M NaCl (peak I) and the second at 0.75 M NaCl (peak II). The L protein in peak I showed significant transcriptional activity in an in vitro transcription reconstitution experiment, whereas the L protein in peak II was inactive. Interestingly, the addition of cytoplasmic extract from uninfected Sf21 cells to peak II completely restored transcription in vitro, indicating the requirement of a host factor(s) for the activity of the L protein. This factor is relatively heat stable and is dissociable from the recombinant L protein. It is also present in BHK, COS, and HeLa cells in detectable levels. The role of the putative host protein(s) in the activation of the L protein is discussed.

Animals↗
PubMed1996

Pneumatic retinopexy: principles and practice.

Pneumatic retinopexy (PR) is an alternative to scleral buckling for the surgical repair of selected retinal detachments. A gas bubble is injected into the vitreous cavity, and the patient is positioned so that the bubble closes the retinal break (s), allowing absorption of the subretinal fluid. Cryotherapy or laser photocoagulation is applied around the retinal break(s) to form a permanent seal. The procedure can be done in an outpatient setting, and no incisions are required. A multicenter randomized controlled clinical trial has demonstrated that the anatomic success rate is comparable to scleral buckling, but the morbidity is significantly less with PR. If the macula was detached for less than two weeks, the visual results are significantly better with PR than with scleral buckling. Cataract surgery was required significantly more often following scleral buckling than following PR. Two independent reports have shown that an attempt with PR does not disadvantage the eye; such that the results of scleral buckling after failed PR are not significantly different than primary scleral buckling. A comprehensive review of the world literature on PR revealed 27 statistical series totaling 1,274 eyes. These combined series had a single-operation success rate of 80%, and 98% were cured with reoperations. Pneumatic retinopexy should be considered in cases without inferior or extensive retinal breaks and without significant proliferative vitreoretinopathy. The cost of buckling varies from 4 to 10 times that of PR.

Air↗
PubMed1995

Role of cellular casein kinase II in the function of the phosphoprotein (P) subunit of RNA polymerase of vesicular stomatitis virus.

The phosphorylation of the P protein of vesicular stomatitis virus by cellular casein kinase II (CKII) is essential for its activity in viral transcription. Recent in vitro studies have demonstrated that CKII converts the inactive unphosphorylated form of P (P0) to an active phosphorylated form P1, after phosphorylation at two serine residues, Ser-59 and Ser-61. To gain insight into the role of CKII-mediated phosphorylation in the structure and function of the P protein, we have carried out circular dichroism (CD) and biochemical analyses of both P0 and P1. The results of CD analyses reveal that phosphorylation of P0 to P1 significantly increases the predicted alpha-helical structure of the P1 protein from 27 to 48%. The phosphorylation defective double serine mutant (P59/61), which is transcriptionally inactive, possesses a secondary structure similar to that of P0. P1, at a protein concentration of 50 micrograms/ml, elutes from a gel filtration column apparently as a dimer, whereas both P0 and the double serine mutant elute as a monomer at the same concentration. Interestingly, unlike wild-type P1 protein, the P mutants in which either Ser-59 or Ser-61 is altered to alanine required a high concentration of CKII for optimal phosphorylation. We demonstrate here that phosphorylation of either Ser-59 or Ser-61 is necessary and sufficient to transactivate L polymerase although alteration of one serine residue significantly decreases its affinity for CKII. We have also shown that P1 binds to the N-RNA template more efficiently than P0 and the formation of P1 is a prerequisite for the subsequent phosphorylation by L protein-associated kinase. In addition, mutant P59/61 acts as a transdominant negative mutant when used in a transcription reconstitution assay in the presence of wild-type P protein.

Amino Acid Sequence↗
PubMed1995

Involvement of cellular casein kinase II in the phosphorylation of measles virus P protein: identification of phosphorylation sites.

The phosphoprotein P gene of measles virus (Edmonston strain) has been cloned in the Escherichia coli expression vector pET-3a with a histidine tag at the C-terminal end. The expressed protein was soluble, unphosphorylated, and constituted 10 to 20% of total cellular protein. Recombinant P protein purified by Ni-affinity chromatography was found to be efficiently phosphorylated in vitro by recombinant casein kinase II (CKII) or by the CKII activity present in the uninfected cell extract. A comparison of phosphopeptide analyses between the in vivo- and the in vitro-32P-labeled P proteins revealed that both proteins share common phosphorylation sites. In an attempt to identify the exact site of the CKII-mediated phosphorylation, we altered specific serine residues located within the CKII consensus motif to alanine by site-directed mutagenesis. The results indicate that Ser 86, Ser 151, and Ser 180 located within the N-terminal half of the P protein are involved in the CKII-mediated phosphorylation of the P protein.

Alkaloids↗
PubMed1995

Casein kinase II is the P protein phosphorylating cellular kinase associated with the ribonucleoprotein complex of purified vesicular stomatitis virus.

Protein kinase activities associated with a highly purified transcriptionally active ribonucleoprotein complex from the virions of vesicular stomatitis virus (VSV) were isolated and characterized. Based upon several biochemical and immunological criteria, the protein kinase activity, which phosphorylated the bacterially expressed unphosphorylated (Po) protein, was shown to be cellular casein kinase II (CKII). These studies included inhibition of the protein kinase by specific inhibitors, phosphorylation of mutant phosphoproteins (P), immunoprecipitation by CKII antibody and Western blot analyses, and finally its ability to activate Po to synthesize RNA in a transcription-reconstitution reaction. The P protein is phosphorylated intracellularly by cellular CKII. The present study demonstrates that VSV specifically packages CKII which remains strongly associated with the ribonucleoprotein complex during morphogenesis.

Amino Acid Sequence↗
PubMed1995

Postsurgical endophthalmitis: diagnosis and management.

Infectious endophthalmitis following intraocular surgery is a complication that could cause severe visual loss or loss of the eye. The categorisation of the event that led to intraocular infection will help the clinician to predict the infectious agent and begin appropriate therapy. Most of the cases of postsurgical endophthalmitis are seen following cataract surgery. It is important for all ophthalmologists, irrespective of specialisation and areas of interest, to be familiar with the management of endophthalmitis. This review briefly describes the facets of clinical and laboratory diagnosis, pathology, and management. While the different viewpoints in the management of endophthalmitis are mentioned in appropriate places, more attention is paid to present a rational approach to the management of endophthalmitis.

Anti-Bacterial Agents↗
PubMed1994

Gh: a GTP-binding protein with transglutaminase activity and receptor signaling function.

The alpha 1-adrenergic receptors activate a phospholipase C enzyme by coupling to members of the large molecular size (approximately 74 to 80 kilodaltons) G alpha h family of guanosine triphosphate (GTP)-binding proteins. Rat liver G alpha h is now shown to be a tissue transglutaminase type II (TGase II). The transglutaminase activity of rat liver TGase II expressed in COS-1 cells was inhibited by the nonhydrolyzable GTP analog guanosine 5'-O-(3-thiotriphosphate) or by alpha 1-adrenergic receptor activation. Rat liver TGase II also mediated alpha 1-adrenergic receptor stimulation of phospholipase C activity. Thus, G alpha h represents a new class of GTP-binding proteins that participate in receptor signaling and may be a component of a complex regulatory network in which receptor-stimulated GTP binding switches the function of G alpha h from transglutamination to receptor signaling.

Amino Acid Sequence↗