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Biomedical subjects

R Raju

Publications and source records attributed to R Raju.

At least 55 records · Page 3Linked to original sources
PubMed1994

Phacoemulsification--a senior surgeon's learning curve.

We sought to prospectively document and evaluate the learning curve of an experienced extracapsular surgeon making a supervised transition from extracapsular cataract extraction (ECCE) to phacoemulsification. Over a period of 2 weeks, 51 phacoemulsification procedures using an endocapsular nucleofractis technique were performed by a single senior surgeon at the Christian Medical College in Vellore, India under the supervision of a visiting US expert with more than 15 years' experience in the technique. Vitreous loss occurred in seven eyes (six prior to the stage of cortical aspiration); failure of capsulorhexis necessitated conversion to standard ECCE in four. Injury to the inferior iris during phacoemulsification was cosmetically significant in three eyes. Two eyes had mild persistent localized corneal edema, but there were no instances of permanent corneal damage. One eye had intraoperative displacement of the nucleus into the vitreous. In one eye with vitreous loss, the implanted intraocular lens dislocated into the vitreous cavity. Two patients had clinically detectable cystoid macular edema at 6 weeks. Eleven patients were lost to follow up after 3 weeks. Six weeks after surgery, 36 of the remaining 40 eyes (90%) had achieved a best-corrected visual acuity of 6/6. We conclude that phacoemulsification requires supervised learning, even for an experienced surgeon. Complications still occurred, but were restricted to the unfamiliar steps of the surgery. Factors identified in the first 2 days of surgery (10 cases) as critical in the smooth transition to phacoemulsification were careful selection of initial cases, a successful capsulorhexis, and hydrodissection with vigorous nucleus mobilization.(ABSTRACT TRUNCATED AT 250 WORDS)

Astigmatism↗
PubMed1993

Percutaneous balloon valvuloplasty in mirror-image dextrocardia and rheumatic mitral stenosis.

A 28-year-old woman with mirror-image dextrocardia and severe rheumatic mitral stenosis underwent successful percutaneous balloon mitral valvuloplasty (PBMV). Standard technique was altered to include transseptal catheterization via the left femoral vein and inter-atrial septal puncture with the transseptal needle rotated to a 7 o'clock position. The predilatation transmitral gradient (mean) of 16 mmHg decreased to 5 mmHg following PBMV. The valve area improved from 1.0 cm2 to 2.6 cm2. There were no complications. This case illustrates that transseptal catheterization can be accomplished safely in patients with unusual cardiac anatomy and mitral valvuloplasty can be performed in patients with mirror-image dextrocardia and rheumatic mitral stenosis.

Adult↗
PubMed1993

Device use patterns and clinical outcome of implantable cardioverter defibrillator patients with moderate and severe impairment of left ventricular function.

The beneficial effects of implanted cardioverter defibrillator (ICD) therapy in patients with malignant ventricular tachyarrhythmias and variable degrees of left ventricular (LV) dysfunction are debated. ICD use and patient survival were examined in 128 patients with malignant ventricular arrhythmias and moderate or severe LV dysfunction. Group I included 64 patients with moderate LV dysfunction (LV ejection fraction of > 30%) and group II, 64 patients with severe LV dysfunction (LV ejection fraction of < or = 30%). Follow-up period ranged from 1 to 78 months. The two groups were similar in age, incidence of coronary artery disease and presenting arrhythmia. The mean LV ejection fraction in group I was 44% +/- 8% and group II was 22% +/- 5% (P < 0.0001). At 4 years of follow-up, 66% of patients from group I and 62% from group II (P = NS) had ICD activation for presumed ventricular tachyarrhythmia. Survival was calculated using actuarial analysis. Arrhythmic or sudden death mortality at 4 years of follow-up was 4% in group I and 7% in group II (P = NS). Cardiac mortality was for group I, 7% (P < 0.05), 12% (P < 0.01), 15% (P < 0.01), and 15% (P < 0.01) for follow-up years 1, 2, 3, and 4, respectively. For group II, cardiac mortality was 27%, 36%, 41%, and 41% for follow-up years for 1, 2, 3, and 4, respectively. The majority of cardiac deaths in both groups was observed in the first 2 years of follow-up.(ABSTRACT TRUNCATED AT 250 WORDS)

Actuarial Analysis↗
PubMed1993

Low-energy endocardial defibrillation using an axillary or a pectoral thoracic electrode location.

BACKGROUND: A significant proportion of patients receiving endocardial defibrillation lead systems must accept either high defibrillation thresholds (DFTs) with lower safety margins or lead implantation by thoracotomy. We examined the feasibility of achieving universal application of endocardial leads and lower defibrillation energy requirements by optimizing the lead system location in conjunction with biphasic shocks. METHODS AND RESULTS: Two defibrillation catheter electrodes were positioned in the right ventricle and superior vena cava. Thoracic patch electrodes were placed at three sites (apical, pectoral, and axillary). Fifteen-joule, 10-J, and 5-J bidirectional simultaneous biphasic shocks were delivered across three different triple electrode configurations (right ventricle, superior vena cava, and patch) after inducing ventricular fibrillation (VF), and DFT was determined. All patients in whom VF was reproducibly inducible (14 patients) could be reproducibly defibrillated at 15 J at one or more patch electrode locations. Fifteen-joule shocks were effective at three thoracic electrode locations in 12 patients and at two electrode locations in 6 patients. The lowest mean single-shock DFT was 8.1 +/- 3.8 J. In 4 patients, ventricular flutter was reproducibly induced and reverted at 15 J in all patients. Mean DFT for the axillary location was 8.3 +/- 3.5 J and was significantly lower than apical (12.8 +/- 5.6 J, P = .008) and pectoral (11.6 +/- 4.1 J, P < .04) patch locations. The probability of success was significantly higher at 10 J with axillary location (78% of patients, P < .03 compared with both other sites) and at 15 J (P < .05 compared with the apical location). Low-energy endocardial defibrillation (< or = 10 J) was feasible in 10 of 14 tested patients at more than 1 thoracic electrode location at 10 J, whereas only 1 of 7 successful patients could be reverted at more than 1 electrode location at 5 J (P < .02). CONCLUSIONS: The use of axillary or pectoral patch lead location can allow endocardial defibrillation with biphasic shocks at energies < or = 15 J in this lead configuration. Virtually universal application of endocardial defibrillation lead systems can be predicted from these data. Reduction in maximum pulse generator output to < or = 25 J using these two thoracic electrode locations with bidirectional shocks can be feasible and maintain an adequate safety margin and permit thoracic pulse generator implantation. Lowering endocardial defibrillation energy < 10 J requires increasing specificity of thoracic electrode location.

Aged↗
PubMed1993

Incidence of complications following 5-fluorouracil with trabeculectomies.

To investigate the incidence of complications reportedly caused by the use of 5-fluorouracil (5-FU) in glaucoma filtering surgery, we performed trabeculectomy with 5-FU on 57 eyes of 56 patients with advanced glaucoma. The mean preoperative intraocular pressure was 34.8 mm Hg. The most frequent complication encountered was superficial punctate keratopathy (26%), followed by shallowing of the anterior chamber (12%), choroidal detachment (9%), and flap retraction (7%). We, however, did not encounter any cases of confluent corneal epithelial defect, subepithelial scarring, or striate melanokeratosis which are usually described with the use of 5-FU. With a mean follow-up of 7.1 months, an intraocular pressure less than 16 mm Hg without hypotensive therapy were achieved in 73.7% of cases, and less than 21 mm Hg without hypotensive therapy in 86%. Considering the low incidence of complications and the high success rate, we recommend the use of 5-FU in routine glaucoma filtering surgery.

Combined Modality Therapy↗
PubMed1993

Anterior capsular support for posterior chamber intraocular lenses following vitreous loss in endocapsular surgery.

We used anterior capsular support for posterior chamber intraocular lenses (PC IOLs) in fourteen eyes with large posterior capsular ruptures. An endocapsular technique preserved the anterior capsule and facilitated implantation. With a median follow up of 8.5 months, all patients had a visual acuity of 6/9 or better. One lens was lost in the vitreous and one patient had a clinically significant cystoid macular edema. In the event of a posterior capsular rupture we suggest this technique as an alternative to anterior chamber or scleral-fixated lenses.

Adolescent↗
PubMed1993

Releasable suture technique for trabeculectomy.

We studied the effect of the releasable suture technique on immediate postoperative intraocular pressure (IOP). Nine eyes of nine patients with glaucoma had trabeculectomy with a releasable suture. In the six eyes that did not receive antimitotics, the suture was released by the fifth postoperative day; in the others suture release was delayed up to the fourteenth day. Of the nine patients, one had an acceptable postoperative IOP and did not need suture release; in another the suture broke and could not be released. In the remaining seven patients, the difference between the pre-release and post-release IOP was statistically significant (p < 0.001). The complications of this technique include failed suture release, subconjunctival hematoma and a distinctive "windshield wiper" keratopathy.

Corneal Diseases↗
PubMed1991

Analysis of Sindbis virus promoter recognition in vivo, using novel vectors with two subgenomic mRNA promoters.

Four types of Sindbis virus vectors, each carrying two promoters for subgenomic mRNA synthesis, were designed to measure relative promoter strengths and to survey potential contextual effects on promoter strengths. One of the promoters in each vector was used as the reference promoter, while the other was the one being tested. We used these vectors to measure the relative strengths of four promoters: the minimal promoter, an extended sequence believed to have full promoter activity, and two mutant promoters, one with an inactivating 3-nucleotide insertion called CR4.1 and the other with a 4-nucleotide deletion called delta 4. The strengths of the promoters were measured by quantitating the RNA transcribed from each promoter in vivo and also by assaying for chloramphenicol acetyltransferase activity encoded by one of the two transcripts. We found that the relative strengths of the promoters were similar in different contexts. The complete promoter was 6-fold more active, the delta 4 promoter was (surprisingly) about twice as active, and the CR4.1 promoter was 100-fold less active than the minimal promoter. At least two contextual effects were identified that can alter the activity of one or both promoters in the vectors. One effect is that given identical promoters, the 3'-proximal promoter on the minus-strand template can be more active than the 5'-proximal promoter. This may be due to preferential association of one or more components of the transcription complex for the 3' end of the minus-strand template. A second effect is promoter competition, particularly when the promoters are closely spaced.

Animals↗
PubMed1990

Nontemplated bases at the 5' ends of Tacaribe virus mRNAs.

Centrifugation of Tacaribe arenavirus-infected cell extracts on CsCl density gradients was used to separate genomes and antigenomes, which band at 1.31 g/ml as nucleocapsids, from mRNAs which pellet. Primer extensions on the banded RNAs showed that the 5' ends of the genomes and antigenomes were unique, whereas primer extensions on the mRNAs showed that their 5' ends were heterogenous in length, extending 0-4 bases beyond the 3' ends of the templates for their synthesis. This suggests that arenavirus mRNAs may initiate by a cap-snatching mechanism, somewhat similar to influenza viruses and bunyaviruses. We also found an extra G residue at the 5' end of the genome RNA, which was not predicted according to current models. This is now the third time that the unexpected G residue has been found at the 5' end of arenavirus genomes.

Animals↗
PubMed1989

The ends of La Crosse virus genome and antigenome RNAs within nucleocapsids are base paired.

The three La Crosse virus genomes are found as circular structures in the electron microscope, and the RNA ends of at least the small (S) and medium (M) segments are highly complementary. When examined for psoralen cross-linking, about half of the S, at most 1 to 2% of the M, and none of the large (L) nucleocapsid RNAs could be cross-linked in virions or at late times intracellularly, under conditions in which each free RNA reacted completely. For the S segment, genomes and antigenomes first detected intracellularly could not be cross-linked at all, and their cross-linkability increased gradually with time. Antigenomes behaved similarly to genomes in all respects. It appears that the majority of all three segments are base paired at their ends and that the limited cross-linkability reflects the accessability of the RNA within nucleocapsids to psoralen. The gradual increase in cross-linkability may be important in persistent mosquito cell infection, in which it correlates with decreased S mRNA synthesis rates, and may be part of the mechanism which this infection becomes self-limiting. The implications of double-stranded RNA panhandles within nucleocapsids are discussed.

Aedes↗
PubMed1989

The translational requirement for complete La Crosse virus mRNA synthesis is cell-type dependent.

The translational requirement to prevent premature termination during La Crosse virus S mRNA synthesis was found to be cell-type dependent. This requirement was present in the BHK, HEL, and Vero cell lines we examined, but not in C6/36 mosquito cells. The cell-dependent translational requirement could be reproduced in vitro by using either cell extracts or purified virions of BHK and C6/36 cells. In the BHK reactions, the polymerase terminated predominantly at nucleotide 175 in the absence of concurrent translation and required translation to read through this position. In the C6/36 reactions, however, the polymerase reads through nucleotide 175 efficiently independent of translation. Reconstitution studies suggested that the translational requirement was due to a factor(s) present in BHK, but not in C6/36, cells.

Animals↗
PubMed1989

La Crosse virus nucleocapsid protein controls its own synthesis in mosquito cells by encapsidating its mRNA.

Within 24 to 48 h of La Crosse virus infection of mosquito cells, greater than 75% of the S mRNA was found to band in CsCl density gradients at the position of genome or antigenome nucleocapsids. The encapsidation of the S mRNA correlates with the repression of N protein synthesis in vivo, and the encapsidated S mRNA cannot be translated in vitro. Unlike genome and antigenome assembly, S mRNA assembly is a relatively slow process, which is not coupled to its synthesis. Within the encapsidated S mRNA population, three forms could be distinguished, those with intact primers which were or were not also assembled with N protein and those in which the primer and up to 3 template bases had been lost. We suggest that genome replication, but not transcription, is down regulated with time in mosquito cells for reasons that are unclear. The pool of unassembled N protein then increased to the point at which it began to interact with its own mRNA, as this mRNA also contains what is considered to be the assembly site, i.e., the conserved sequences at the 5' ends of all genome and antigenome chains. This lead to the assembly of the entire mRNA, except for the nontemplate primer. Some of the primers were then also assembled with N protein, whereas others were digested to produce truncated mRNAs.

Animals↗
PubMed1989

A cis-acting mutation in the Sindbis virus junction region which affects subgenomic RNA synthesis.

The synthesis of Sindbis virus minus-strand and genomic and subgenomic RNAs is believed to require specific cis-acting sequences or structures in the template RNAs and a combination of virus-specific proteins and host components which act in trans. A conserved sequence of about 21 nucleotides in the junction region and encompassing the start site for the subgenomic RNA has been proposed to function as the promoter on the minus-strand template for synthesis of the subgenomic RNA (J.-H. Ou, C. M. Rice, L. Dalgarno, E. G. Strauss, and J. H. Strauss, Proc. Natl. Acad. Sci. USA 79:5235-5239, 1982). We introduced a three-base insertion in this sequence, which also inserts a single amino acid near the COOH terminus of nsP4, in a cDNA clone of Sindbis virus from which infectious RNA transcripts can be generated. The phenotype of this mutant, called Toto1100CR4.1, was studied after RNA transfection of chicken embryo fibroblasts or BHK cells. The mutation leads to a drastic reduction in the level of the subgenomic RNA but does not alter the start site of the RNA. Probably as a consequence of depressed structural-protein synthesis, very few progeny virions are released and the mutant makes tiny or indistinct plaques even after prolonged incubation. The cis-acting effect of this mutation was demonstrated by incorporating either a wild-type or mutant junction region into a defective-interfering RNA and examining the relative synthesis of defective-interfering RNA-derived subgenomic RNA in vivo in the presence of wild-type helper virus. These results show that the junction region is recognized by yet unidentified viral trans-acting components for subgenomic RNA synthesis. When the Toto1100CR4.1 mutant was passaged in culture, plaque morphology variants readily arose. A total of 24 independent revertants were isolated, and 16 were characterized in detail. All revertants analyzed showed an increase in the level of subgenomic RNA synthesis. Sequence analysis of the junction region showed that all were pseudorevertants, with only two containing potentially compensating changes in the junction region. An assay was developed to identify revertants with second-site changes in trans-acting viral components involved in subgenomic RNA synthesis. At least two such revertants were identified. Mapping of these and other second-site compensating mutations may provide genetic clues as to which virus-specific protein(s) is responsible for interaction with the conserved junction region to promote subgenomic RNA synthesis.

Amino Acid Sequence↗
PubMed1988

LaCrosse virus gene expression in mammalian and mosquito cells.

LaCrosse virus infection of mammalian BHK cells is highly cytopathic, whereas that of mosquito C6/36 cells is asymptomatic and persistent. When the individual mRNAs and their genome segments are followed in parallel infections, cytopathic effects were found to correlate with the rate of synthesis, but not the accumulation, of the viral RNAs. The change from the acute to the persistent phase of the infection in C6/36 cells was found to take place at 24 hr p.i., at which time genome and N protein synthesis was severely reduced, even though mRNA levels remained high. When the persistent infection was followed for 72 days, the total amounts of genomes and their relative proportions were found to fluctuate greatly, whereas mRNA levels were either severely reduced or undetectable. DI genomes could not be detected during this time. The self-limiting nature of the mosquito cell infection appears to be due the translational control of N protein synthesis.

Animals↗
PubMed1988

La Crosse virus infection of mammalian cells induces mRNA instability.

La Crosse virus infection of BHK cells leads to a dramatic shutoff of not only host protein synthesis but also viral protein synthesis later in infection. This shutoff can be accounted for by the loss of the cytoplasmic cellular and viral mRNAs. The induction of mRNA instability requires extensive virus replication, since when cycloheximide is added early in infection the preexisting viral and cellular mRNAs do not decrease upon incubation of the cultures. Pretreatment of the cultures with actinomycin D does not affect the ability of La Crosse virus infection to induce mRNA instability, and examination of the rRNAs shows no evidence of specific degradation due to activation of the interferon-associated latent RNase. The induction of mRNA instability therefore does not appear to operate through an interferon pathway. Viral mRNA synthesis, on the other hand, is not turned off during infection, and the cap-dependent endonuclease involved in viral mRNA initiation may be responsible for the mRNA instability.

2',5'-Oligoadenylate Synthetase↗
PubMed1987

Translational requirement of La Crosse virus S-mRNA synthesis: in vitro studies.

The exceptional requirement of La Crosse virus mRNA synthesis for ongoing protein synthesis in vivo was examined in vitro by using purified virions and a reticulocyte lysate. Transcription from the S genome produced two incomplete transcripts (110 and 205 nucleotides [nt]) in the absence of the lysate, whereas S-mRNA (900 nt) was predominantly made when the lysate was present. The addition of drugs which inhibit protein synthesis also inhibited the synthesis of S-mRNA, and in some cases led to the reappearance of the 205-nt RNA. Reconstruction experiments demonstrated that the incomplete transcripts were not the result of rapid and selective degradation of S-mRNA but were due to premature termination of the polymerase at defined sites. The requirement for ongoing protein synthesis for productive transcription in vitro is not at the level of chain initiation but for elongation of the nascent RNA beyond these sites.

Animals↗
PubMed1987

Translational requirement of La Crosse virus S-mRNA synthesis: in vivo studies.

By using methods to isolate cytoplasmic RNAs which limit degradation, the effect of drugs which inhibit protein synthesis on the accumulation of La Crosse virus plus-strand S RNAs in vivo has been studied. Cycloheximide and puromycin treatment of infected cultures caused an abortive transcript of ca. 205 nucleotides (nt) to accumulate, whereas pactamycin led to the appearance of an RNA which was slightly shorter (ca. 200 nt). Both the 205- and 200-nt RNAs contained the same range of host primers at their 5' end, but their 3' ends mapped at ca. positions 175 and 165, respectively. Examination of the sequence in this region and at the mature mRNA termination site (position 886) suggests that the sequence YAAAAAT(A)GCAG is involved in transcription termination.

Animals↗