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Biomedical subjects

P Joshi

Publications and source records attributed to P Joshi.

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

Crystallization of a fragment of human fibronectin: introduction of methionine by site-directed mutagenesis to allow phasing via selenomethionine.

Crystals of a fragment of human fibronectin encompassing the 7th through the RGD-containing 10th type III repeats (FN7-10) have been produced with protein expressed in E. coli. The crystals are monoclinic with one molecule in the asymmetric unit and diffract to beyond 2.0 A Bragg spacings. A mutant FN7-10 was produced in which three methionines, in addition to the single native methionine already present, have been introduced by site-directed mutagenesis. Diffraction-quality crystals of this mutant protein have been grown in which methionine was replaced with selenomethionine. The introduction of methionine by site-directed mutagenesis to allow phasing from selenomethionyl-substituted crystals is shown to be feasible by this example and is proposed as a general approach to solving the crystallographic phase problem. Strategies for selecting propitious sites for methionine mutations are discussed.

Amino Acid Sequence↗
PubMed1994

Decreased insulin-like growth factor I-mediated protein tyrosine phosphorylation in human olivopontocerebellar atrophy and lurcher mutant mouse.

We examined insulin-like growth factor I (IGF-I)-dependent phosphorylation and protein tyrosine kinase (PTK) activity in cerebellar cortex of normal humans, patients with olivopontocerebellar atrophy (OPCA) ("C" kindred) and in lurcher mutant mouse, a suggested animal model for OPCA. PTK activity and IGF-I-dependent protein tyrosine phosphorylation was significantly reduced in cerebellar cortex of human OPCA patients as compared to the normal controls. Immunoblot analysis also demonstrated a decrease in cerebellar 80 kDa phosphotyrosine protein in these patients. By autoradiography, IGF-I receptors were localized in the molecular layer of 30-day-old control and lurcher mutant mice cerebella. However, the lurcher mutant mice showed a decrease in [125I]-IGF-I binding in the molecular layer as compared to the littermate controls. The IGF-I receptor autophosphorylation was also markedly reduced in 15-day- and 22-day-old lurcher cerebella. These results suggest that the process of cerebellar degeneration in human OPCA and lurcher mutant mouse may be associated with altered IGF-I receptor binding and protein tyrosine phosphorylation.

Amino Acid Sequence↗
PubMed1994

Trial of intravesical versus extravesical ureteroneocystostomy in renal transplant recipients.

We carried out a trial to evaluate the complication rate of intravesical (LP) versus extravesical (Lich) ureteroneocystostomy in recipients of renal transplantation. Ureteric stenosis was the predominant complication in the LP technique, which was more difficult to correct. Complications by the Lich technique were urinary leaks, which were managed successfully by prolonged bladder drainage. We conclude that the Lich technique is simpler to perform and avoids the complication of ureteric stenosis, and should therefore be the procedure of choice for ureteric implantation in recipients of renal transplants.

Adult↗
PubMed1994

Cell cycle-dependent inhibition of p34cdc2 synthesis by transforming growth factor beta 1 in cycling epithelial cells.

Cycling epithelial cells were shown to reversibly arrest in late G1 phase following treatment with transforming growth factor beta 1. Associated with this G1-S phase arrest was a decrease in the synthesis and histone H1 kinase activity of p34cdc2. Transforming growth factor beta 1 did not reduce p34cdc2 levels by modulating the turnover of newly synthesized p34cdc2. The decrease in p34cdc2 synthesis preceded any detectable effect on DNA synthesis. Moreover, the action of transforming growth factor beta 1 was regulated in a cell cycle-specific manner; epithelial cells were sensitive to transforming growth factor beta 1 only during the G1 phase. The results suggest that p34cdc2 might be a useful biochemical marker for investigating the mechanism(s) of transforming growth factor beta 1 signaling.

Animals↗
PubMed1993

Cell- and heparin-binding domains of the hexabrachion arm identified by tenascin expression proteins.

We have produced a set of bacterial expression proteins corresponding to 10 segments of tenascin and two of fibronectin and tested them for heparin binding and cell adhesion. We used polymerase chain reaction cloning to terminate the segments precisely at domain boundaries. Heparin binding activity was mapped to two different tenascin segments: one comprising the fourth and fifth fibronectin type III domains, and to TNfbg, the fibrinogen-like terminal knob. TNfbg, but none of the other tanascin segments, also supported adhesion of primary rat embryo skin fibroblasts. The fibroblasts did not spread on TNfbg but remained rounded. Cell binding to TNfbg occurred in the presence or absence of divalent cations and was not inhibited by RGD peptides, suggesting that integrins are not involved. Fibroblast binding to TNfbg was strongly inhibited by soluble heparin, by treating the cells with heparitinase, or by culture conditions that cause undersulfation of proteoglycans. These observations suggest that cell attachment to TNfbg is mediated by cell surface proteoglycans. We have also made full-length cDNA constructs for the largest and smallest splice variants of human tenascin, as well as one truncated after the 14th epidermal growth factor-like domain, in the pNUT mammalian cell expression vector. Stably transfected baby hamster kidney cell lines secreted large quantities of tenascin, and this was assembled into normal hexabrachions, the arm length corresponding to the construct.

Animals↗
PubMed1993

Characterization of paralogous and orthologous members of the superoxide dismutase gene family from genera of the halophilic archaebacteria.

Four species representing three genera of halophilic archaebacteria were examined for the presence of genomic sequences that encode proteins of the superoxide dismutase family. Three species, Halobacterium cutirubrum, Halobacterium sp. strain GRB, and Haloferax volcanii, contain duplicated (paralogous) genes of the sod family; a fourth species, Haloarcula marismortui, contains only a single gene. These seven genes were cloned and sequenced, and their transcripts were characterized by Northern (RNA) hybridization, S1 nuclease protection, and primer extension. The expression of one of the two genes in H. cutirubrum, Halobacterium sp. strain GRB, and Haloferax volcanii was shown to be elevated in the presence of paraquat, a generator of superoxide radicals. The other genes, including the single gene from Haloarcula marismortui, exhibited no elevated expression in the presence of paraquat. The 5' and 3' flanking regions of all the genes contain recognizable promoter and terminator elements that are appropriately positioned relative to the 5' and 3' transcript end sites. Between genera, the orthologous paraquat-responsive genes exhibit no sequence similarity in either their 5' or 3' flanking regions, whereas the orthologous nonresponsive genes exhibit limited sequence similarity but only in the 5' flanking region. Within the coding region, the two paralogous genes of Haloferax volcanii are virtually identical (99.5%) despite the absence of similarity in the flanking regions. In contrast, the paralogous genes of H. cutirubrum and Halobacterium sp. strain GRB are only about 87% identical. In the alignment of all seven sequences, there are nine codon positions where both the TCN and AGY serine codons are utilized; some or all of these may well be examples of convergent evolution.

Amino Acid Sequence↗
PubMed1993

Structure, function, and evolution of the family of superoxide dismutase proteins from halophilic archaebacteria.

The protein sequences of seven members of the superoxide dismutase (SOD) family from halophilic archaebacteria have been aligned and compared with each other and with the homologous Mn and Fe SOD sequences from eubacteria and the methanogenic archaebacterium Methanobacterium thermoautotrophicum. Of 199 common residues in the SOD proteins from halophilic archaebacteria, 125 are conserved in all seven sequences, and 64 of these are encoded by single unique triplets. The 74 remaining positions exhibit a high degree of variability, and for almost half of these, the encoding triplets are connected by at least two nonsynonymous nucleotide substitutions. The majority of nucleotide substitutions within the seven genes are nonsynonymous and result in amino acid replacement in the respective protein; silent third-codon-position (synonymous) substitutions are unexpectedly rare. Halophilic SODs contain 30 specific residues that are not found at the corresponding positions of the methanogenic or eubacterial SOD proteins. Seven of these are replacements of highly conserved amino acids in eubacterial SODs that are believed to play an important role in the three-dimensional structure of the protein. Residues implicated in formation of the active site, catalysis, and metal ion binding are conserved in all Mn and Fe SODs. Molecular phylogenies based on parsimony and neighbor-joining methods coherently group the halophile sequences but surprisingly fail to distinguish between the Mn SOD of Escherichia coli and the Fe SOD of M. thermoautotrophicum as the outgroup. These comparisons indicate that as a group, the SODs of halophilic archaebacteria have many unique and characteristic features. At the same time, the patterns of nucleotide substitution and amino acid replacement indicate that these genes and the proteins that they encode continue to be subject to strong and changing selection. This selection may be related to the presence of oxygen radicals and the inter- and intracellular composition and concentration of metal cations.

Amino Acid Sequence↗
PubMed1993

Endothelial cells adhere to the RGD domain and the fibrinogen-like terminal knob of tenascin.

We have found that endothelial cells adhere much more strongly than fibroblasts to domains of tenascin and fibronectin. Endothelial cells adhered weakly, without spreading, to bacterial expression proteins corresponding to the tenth fibronectin type III (FN-III) domain of fibronectin, which contains the RGD. A larger fibronectin protein, containing this domain and the three amino-terminal 'synergy' domains gave strong adhesion and spreading. Two widely separated domains of tenascin gave adhesion. The third FN-III domain, TNfn3, which contains an RGD sequence in human and chicken tenascin, gave very strong adhesion and spreading of endothelial cells when tested as an isolated domain. Larger segments containing TNfn3 and the adjacent TNfn2 gave weaker adhesion, probably because the RGD sequence is partially blocked. Adhesion to this domain required divalent cations, was exquisitely sensitive to soluble GRGDSP peptide, and was blocked by antisera to the integrin alpha v beta 3. The second tenascin adhesion domain was the fibrinogen-like C-terminal knob, TNfbg. Cells adhered to but did not spread on this domain. This adhesion required divalent cations and was also sensitive to GRGDSP peptide, so it may be mediated by an integrin receptor. We have explored a range of conditions for preparing the adhesion substratum, and our results may resolve the controversy over whether tenascin can act as a substratum adhesion molecule. When coated for short times (1-2 hours) on plastic, tenascin had no adhesion activity, in contrast to fibronectin and the expression proteins, which gave strong adhesion under these conditions. When coated for longer times (12-24 hours) on plastic, the tenascin substratum supported good adhesion, but not spreading, of endothelial cells. Tenascin coated on nitrocellulose gave substantially stronger adhesion than on plastic, but still required long coating times for maximal activity. Adhesion of endothelial cells to native TN was inhibited by GRDGSP peptide. The cell adhesion activity demonstrates the presence on endothelial cells of tenascin receptors, which may play a supportive role in angiogenesis, in the structure of blood vessels, or in binding tenascin to the cell surface to elicit or enhance a signalling function.

Amino Acid Sequence↗
PubMed1992

Successful renal transplantation in a patient with systemic sarcoidosis and renal failure due to focal glomerulosclerosis.

In 1987, a patient presented with pulmonary sarcoidosis and progressive renal failure. Percutaneous renal biopsy showed focal glomerular sclerosis (FGS). Over the subsequent 4-year period, her renal failure progressed to require hemodialysis. During this time, her pulmonary sarcoidosis was treated with daily corticosteroid therapy and she remained in clinical remission. The patient received a one-haplotype-identical living-related renal transplant from her mother. The patient is now 25 months posttransplant with a serum creatinine of 106 mumol/L (1.2 mg/dL). Her sarcoidosis continues to be in clinical remission. In this report, we review the rare association of a primary glomerular lesion causing renal failure in patients with sarcoidosis. In addition, we detail the first successful renal transplant in such a patient.

Adult↗
PubMed1992

A high flow semi-open system for preoxygenation: an evaluation.

We have compared an alternative breathing system for preoxygenation comprising a Hudson face mask with high oxygen inflow (48 litre min-1) and a Mapleson A breathing system (100 ml kg-1 min-1). The study consisted of two parts: the first involved adult volunteers (10 male, seven female) and the second part used a lung model for spontaneous ventilation with a sinusoidal ventilatory wave pattern. In the volunteers, preoxygenation was achieved at mean times of 138 (SD 31.3) s and 164 (SD 36.7) s with the high flow semi-open and Mapleson A systems, respectively. In the lung model, at peak inspiratory flow rates of 30 and 40 litre min-1, the preoxygenation times were 139 and 120 s, respectively, with the semi-open system and 167 and 156 s with the Mapleson A system. The high flow semi-open system may be an alternative for current techniques, provided peak inspiratory flows are not excessive.

Adult↗
PubMed1992

Nitrous oxide administration using commonly available oxygen therapy devices.

Administration of nitrous oxide is useful for providing sedation and analgesia. The therapeutic range for nitrous oxide is 20-30%. Several oxygen treatment devices have been used for administering nitrous oxide, but little is known about the concentrations of nitrous oxide and oxygen delivered to the trachea. We have studied this, using an analogue lung model, with several oxygen therapy devices. With a 1:1 nitrous oxide-oxygen mixture in the primary flow for all systems, end-expired nitrous oxide concentrations varied between 6.5% and 34.3%. Therapeutic concentrations were produced using the Hudson (nominal oxygen concentration 60%) fixed-performance mask, the variable performance Hudson mask at 4 litre min-1, the MC masks at 4 and 6 litre min-1 and the nasal prongs at 6 and 8 litre min-1. Simultaneous end-expired oxygen concentrations for all devices tested were within a safe range.

Analgesia↗
PubMed1992

An evaluation of oxygen delivery using nasal prongs.

Oxygen delivery using nasal prongs was assessed using a lung model for spontaneous ventilation. The analogue lung was attached to a manikin, which provided a model of the 'face and pharynx' to which the nasal prongs were applied. Oxygen concentrations were measured in the model trachea at varying fresh gas inflow and peak inspiratory flows. The study demonstrated enormous variability in the both the peak-inspired (26.3-90.0%) and end-expired concentrations (25.2-78.6%) of oxygen delivered to the trachea. There was a regular relationship between the ratio of peak inspiratory flows, expressed over fresh gas inflow and the end-expired oxygen concentrations which could allow estimation of inspired oxygen concentration.

Evaluation Studies as Topic↗
PubMed1992

Pre-oxygenation: the Hudson mask as an alternative technique.

The use of a simple oxygen facemask (Hudson) with high oxygen inflow (48 l.min-1) was investigated as a technique for pre-oxygenation, comparing it with the Magill system (oxygen flow: 100 ml.kg-1.min-1). One hundred and thirty-eight patients scheduled for elective gynaecological and orthopaedic surgery were studied: group 1, Hudson mask and group 2, Magill system (ASA 1-2, n = 107); group 3, Hudson mask and group 4, Magill system (ASA 3, n = 30). Pre-oxygenation was assessed by measuring the times to 97%, 95% and 93% arterial desaturation (finger pulse oximetry) following 3 min of pre-oxygenation. The times taken to achieve these end-points in all the study groups suggest that the Hudson mask offers an alternative technique for pre-oxygenation.

Adult↗
PubMed1992

Immunization status of casualty attenders: risk factors for non-compliance and attitudes to 'on the spot' immunization.

Outbreaks of vaccine preventable infections have focused attention on 'missed opportunities' for immunizing children. The immunization status of 520 consecutive children attending Casualty during a 10 day period was studied. Only 70% of children had received their diphtheria, tetanus, pertussis (DTP) and poliomyelitis immunization at the appropriate time, 13% had completed the schedule later than recommended and 17% had immunizations overdue by 4 weeks or more. For measles (mumps/rubella) vaccine (MM or MMR) 75% were up to date, 10% were given late and 15% were overdue. A subset of 171 families was interviewed to evaluate factors affecting compliance. Families possessing a Social Security 'Health Care Card', whose father was unemployed, who spoke poor English or who had lived in Australia for 5 years or less were significantly more likely (P < 0.02) to be inadequately immunized. There were 84 children whose immunization was overdue and who were well enough to be immunized. The parents of 70 (83%) of these 84 said that they would agree to 'on the spot' immunization if it were available; 14 (17%) parents refused, the commonest reason for refusal being that the parents felt that the child was too sick at the time to be immunized.

Child↗
PubMed1992

Nitrous oxide-oxygen analgesia: the performance of the MC mask delivery system.

Nitrous oxide administration via an oxygen facemask such as the Mary Catterall (MC) is an accepted technique for supplementing regional anaesthesia. This study assessed the range of nitrous oxide and oxygen concentrations delivered to the trachea using a mechanical lung model. The inspired concentrations of these two gases were determined by the total fresh gas inflow rates, their concentrations in the fresh gas mixture and the peak inspiratory flow rate of the analogue lung. A 1:1 nitrous oxide-oxygen mixture in the fresh gas with an inflow rate of 4-6 l/min is recommended to ensure adequate oxygen delivery into the trachea as well as achieving the therapeutic concentrations of nitrous oxide (20-30%).

Analgesia↗
PubMed1992

First and second metatarsophalangeal joint dislocation. A case report.

A case of dislocated first and second metatarsophalangeal joints was reported along with the mechanics and mechanism of injury. The practitioner must be familiar with the mechanism of injury and radiographic classification to determine the proper indications for closed versus open reduction. The authors' review of the literature did not reveal a similar case involving a dislocation of the first and second metatarsophalangeal joints.

Bandages↗