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Biomedical subjects

S A Desai

Publications and source records attributed to S A Desai.

16 recordsLinked to original sources
PubMed1998

Characterization of human anti-high molecular weight-melanoma-associated antigen single-chain Fv fragments isolated from a phage display antibody library.

The human high molecular weight-melanoma-associated antigen (HMW-MAA) meets the criteria to be used as an immunogen for immunotherapy of malignant melanoma, because it is expressed by a large percentage of melanoma lesions with limited heterogeneity and has a restricted distribution in normal tissues. The high immunogenicity of the HMW-MAA in BALB/c mice has resulted in the development of a large number of anti-HMW-MAA monoclonal antibodies (mAbs). In contrast, no human anti-HMW-MAA mAbs have been described. Because the latter may serve as useful probes to characterize the antigenic profile of the HMW-MAA, human anti-HMW-MAA single-chain fragments of the variable region (scFvs) were isolated by panning synthetic scFv library 1 on purified HMW-MAA. Colony hybridization studies and nucleotide sequence analysis revealed that scFv 19, 44, 56, and 61 belong to the V(H)3 gene family and use the DP-38 germ-line gene segment but have a diverse third complementarity-determining region. The human scFvs share some characteristics with mouse anti-HMW-MAA mAb but also display some distinct features. Like mouse mAbs, human scFvs recognize determinants of HMW-MAA with a heterogeneous cellular and molecular distribution in human melanoma cells. Furthermore, like some mouse mAbs, human scFvs react with rat neural cells expressing the chondroitin sulfate proteoglycan NG2, which shows 81% homology to the HMW-MAA. However, at variance with mouse mAbs, the human scFvs show poor reactivity with guinea pig melanoma cells. Lastly, human scFv 61 stains both benign and malignant lesions of melanocytic origin, although with a lower frequency than mouse mAbs. Analysis of the clinical significance of the differential expression of the scFv 61-defined determinant in melanoma lesions will be facilitated by its reactivity with formalin-fixed melanoma lesions. In contrast to mouse mAb, scFv 61 immunoprecipitates the >450-kDa chondroitin sulfate proteoglycan component of the HMW-MAA, but not its 250-kDa subunit from melanoma cells. Thus, contrary to the current view about the structure of HMW-MAA, our results demonstrate that the two components are not associated. The described scFv antibodies, which represent the first example of human anti-HMW-MAA antibodies, have provided novel information about the structure of this antigen. Future studies will assess the impact of these in vitro-assembled antibody fragments on the identification of antigenic determinants of the HMW-MAA that can be recognized by the human immune system.

Animals↗
PubMed1998

Adverse neurodevelopmental outcome after extracorporeal membrane oxygenation among neonates with bronchopulmonary dysplasia.

OBJECTIVE: The relationship between bronchopulmonary dysplasia (BPD) and neurodevelopmental outcome after extracorporeal membrane oxygenation (ECMO) has not been extensively reported. We compared the outcomes in a large series of infants with and without BPD after ECMO. STUDY DESIGN: Hospital charts and follow-up records of 145 infants treated with ECMO (1985 through 1990) were reviewed. Complete long-term respiratory and follow-up outcome data were available in 64 infants. BPD occurred in 17 survivors; the remaining 47 did not have BPD. RESULTS: Babies with BPD were more likely to have had respiratory distress syndrome. Mean (+/- SD) age at ECMO initiation was later for the BPD group (127+/-66 vs 53+/-39 hours, p < 0.001), and the duration of ECMO treatment was longer (192+/-68 vs 119+/-53 hours, p < 0.001). Bayley Scales of Infant Development scores at <30 months were lower in infants with BPD (p < 0.001), as were three of four Mullen Scales of Early Learning scores (> or = 30 months, p < 0.001 or p = 0.01). At 57+/-16 months 11 (64%) patients with BPD had mild neurologic disabilities, and 3 (18%) had severe disabilities. At a similar age (53+/-16 months, p = NS) 16 (34%) patients without BPD had mild disabilities, whereas 2 (4%) had severe disabilities (p < 0.01). CONCLUSIONS: The occurrence of BPD after ECMO is associated with adverse neurodevelopmental outcome. Patients with BPD after ECMO merit close long-term follow-up.

Bronchopulmonary Dysplasia↗
PubMed1997

Pore size of the malaria parasite's nutrient channel.

The malaria parasite, Plasmodium falciparum, requires large amounts of nutrients to sustain its rapid growth within the human red blood cell. A recently identified ion channel on the surface of the intraerythrocytic parasite may provide direct access to these nutrients in the red blood cell cytosol. Evidence supporting this role was obtained by incorporating this channel into planar lipid bilayers. In bilayers, this channel has conductance and gating properties identical to the in situ channel, passes soluble macromolecules of up to 1400 Da, and functions as a high capacity, low affinity molecular sieve. These properties, remarkably similar to those of a pore on Toxoplasma gondii (another protozoan parasite causing human disease), suggest a novel class of channels used by these intracellular parasites to acquire nutrients from host cytosol.

Animals↗
PubMed1996

A novel pathway for Ca++ entry into Plasmodium falciparum-infected blood cells.

Growth of the human malaria parasite, Plasmodium falciparum, within the red blood cell (RBC) requires external Ca++ and is associated with a markedly elevated intracellular Ca++ concentration, [Ca++]i. We used 45Ca++ flux studies and patch clamp recordings to examine the mechanisms responsible for this increased [Ca++]i. The 45Ca++ flux studies indicated that net Ca++ entry into parasitized RBCs (PRBCs) is 18 times faster than into unparasitized ATPase that keeps the [Ca++]i of unparasitized RBCs exceedingly low. Acceleration of the preexisting Ca++ entry, ATPase that keeps the [Ca++] of unparasitized RBCs exceedingly low. Acceleration of the preexisting Ca++ entry, mediated by a divalent cation carrier, also cannot explain Ca++ accumulation in PRBCs: there are fundamental differences in substrate preference and in the effects of external Ca++ on 45Ca++ efflux between unparasitized RBCs and PRBCs. Patch clamp of intact PRBC surface membranes revealed rare unitary channel openings not observed on unparasitized RBCs. With 80 mM of CaCl2 in the patch pipette, this channel carried inward current, suggesting Ca++ entry at a rate comparable with the observed 45Ca++ flux. These data indicate that the malaria parasite induces a novel pathway in the host RBC membrane for Ca++ entry and suggest that this pathway is a Ca++-permeable channel.

Animals↗
PubMed1994

Right common carotid artery reconstruction after extracorporeal membrane oxygenation: vascular imaging, cerebral circulation, electroencephalographic, and neurodevelopmental correlates to recovery.

OBJECTIVE: Right common carotid artery (RCCA) ligation after extracorporeal membrane oxygenation by venoarterial bypass may contribute to lateralized cerebral injury. Reconstruction of this artery after extracorporeal membrane oxygenation has proved feasible but has not been evaluated for neurologic outcome in any substantial series of infants. METHODS: We evaluated RCCA reconstruction in 47 infants treated with ECMO and compared their cerebrovascular and neuroanatomic imaging findings, electroencephalograms, and developmental outcomes with those of 93 infants who had no reconstruction. SUMMARY RESULTS: Color Doppler blood flow imaging revealed that carotid artery patency was usually obtained after RCCA reconstruction. Right internal carotid and bilateral anterior and middle cerebral arterial blood flow velocities were generally higher, and were more symmetrically distributed in infants with reconstructed RCCA. Electroencephalography did not disclose an increased risk of deterioration or marked abnormalities in infants after reconstruction, nor were neuroimaging findings consistent with an increased number of either focal or generalized abnormalities. Neurodevelopmental follow-up revealed no differences in the incidence of delays between those with a reconstructed RCCA and those with a ligated RCCA during the first year of life. CONCLUSIONS: Reconstruction of the RCCA after extracorporeal membrane oxygenation may facilitate normal distribution of cerebral blood flow through the circle of Willis, and may augment both left and right middle cerebral artery blood flow immediately after decannulation. The long-term consequences of either ligation or reconstruction of the RCCA will require careful scrutiny, however, before either course is recommended routinely.

Blood Flow Velocity↗
PubMed1993

A nutrient-permeable channel on the intraerythrocytic malaria parasite.

During its 48-hour cycle inside the red blood cell, the human malaria parasite, Plasmodium falciparum, increases its volume 25-fold and divides asexually. This rapid growth demands large amounts of nutrients, a problem exacerbated by the lower metabolic rate and relative ionic impermeability of the host red blood cell. Direct passage of small nutrients across the two membranes that separate the parasite from the erythrocyte cytosol may be important for parasite development and has been demonstrated for radiolabelled glucose, amino acids, and purine nucleosides. Flux studies on plasmodia are limited, however, to suspensions of erythrocyte-free parasites and so cannot be used to examine the individual transport properties of the two membranes involved. Here we use the cell-attached patch clamp method to overcome this limitation. After removing the intervening red blood cell membrane and forming gigaohm seals on the small (3-5 microns) parasite, we studied transport across the parasitophorous vacuole membrane (PVM), the outer of the two membranes that separate the parasite from the erythrocyte cytosol. A 140-pS channel which is permeable to both cations and anions was identified on the PVM. This channel is present at high density, is open more than 98 per cent of the time at the resting potential of the PVM, and is permeable to lysine and glucuronate. The channel can readily transport amino acids and monosaccharides across the PVM and may be essential for fulfilling the parasite's metabolic demands.

Animals↗
PubMed1993

Modulation of tumour associated antigen expressed on human squamous cell carcinoma cell lines by recombinant interferon-alpha.

Our earlier studies have shown that monoclonal antibody (Mab) 3F8E3 generated against a head and neck cancer cell line LICR-LON-HN2 showed reactivity with squamous cell carcinoma (SCC) irrespective of the tissue of origin and identified antigens on SCC cell lines AW 13516 and AW 8507. The affinity constants (Ka) for binding of Mab 3F8E3 to AW 13516 and AW 8507 cell lines were 6.2 x 10(8) and 4.3 x 10(8) mol/l and it identified 6.8 x 10(4) and 3.77 x 10(4) sites/cell, respectively, as determined by Scatchard analysis. Treatment of both the cell lines with recombinant human interferon-alpha (rHu-IFN alpha) increased the binding affinity of the Mab but did not increase the number of binding sites on the SCC cell lines. Shedding of antigen recognised by the Mab in the culture supernatant of the cell lines was reduced after rHu-IFN alpha treatment. The results suggest that rHu-IFN alpha may bring about a firm anchorage of the tumour associated antigen on the SCC cells. Cells modulated with rHu-IFN alpha may serve as better targets for assessing cell mediated as well as Mab mediated cytotoxicity in oral cancer patients.

Animals↗
PubMed1993

Modulatory effect of cytokines on natural killer and antibody dependent cellular cytotoxicity directed to squamous cell carcinoma targets by lymphocytes from oral cancer patients.

Cells from solid tumours are generally poor targets for natural killer (NK) cytotoxicity and antibody dependent cellular cytotoxicity (ADCC). In this paper, we have analysed NK cytotoxicity and ADCC mediated by peripheral blood mononuclear cells from healthy individuals and oral cancer patients before and after modulation with recombinant interleukin-2 (rIL-2), on target cells derived from two squamous cell carcinoma (SCC) cell lines prior to and after treatment with recombinant interferon-alpha (rIFN alpha). Target SCC cell directed monoclonal antibody 3F8E3 was used in ADCC. The results showed that the unmodulated SCC cells were poor targets for NK and ADCC compared to standard targets (K562 cells and chicken red blood cells, respectively). Modulation of targets alone with rIFN alpha showed moderate increase in their susceptibility while rIL-2 treated effectors could significantly lyse even unmodulated targets. Combined treatment of targets with rIFN alpha and effectors with rIL-2 showed additive enhancement in NK and ADCC activity against SCC cells. Lymphocytes from treated patients with recurrent disease could not efficiently lyse SCC targets even after combined modulation.

Adult↗
PubMed1993

Cardiac involvement in systemic sclerosis.

The clinical profile of systemic sclerosis with screening for cardiac involvement and the effect of cold stimulation on myocardium are described in 17 patients. Besides routine investigations, echocardiography, radionuclide ventriculography and computerized stress test, both before and after cold pressor test were done. Ten patients had abnormal ECG findings, two patients had mild pericardial effusion and five patients had moderate grade pulmonary hypertension. Echocardiography and radionuclide ventriculography did not reveal wall motion abnormality either before or after cold stimulation. Computerized stress test was positive for ischaemia following cold stimulation in one patient.

Adult↗
PubMed1992

Right common carotid artery reconstruction in neonates after extracorporeal membrane oxygenation: color Doppler imaging.

Thirty-three neonates treated for reversible respiratory failure underwent reconstruction of their previously ligated right common carotid arteries (RCCAs) immediately after extracorporeal membrane oxygenation (ECMO). Cerebral color Doppler imaging, performed during and repeatedly after ECMO, revealed antegrade flow in the right internal carotid artery (ICA) in all neonates within 6 days after successful RCCA reconstruction. Mean ICA velocity was significantly less in the right artery compared with the left during ECMO and within 1 hour of reconstruction, but there was no difference after 12 hours. In neonates with successful RCCA reconstruction, the flow in the proximal right anterior cerebral artery was antegrade in only 4% of examinations during ECMO but became antegrade in 94% after 6 days. Retrograde flow in the right posterior communicating artery persisted in 50% of examinations performed 1 day after reconstruction. ICA flow become antegrade with symmetric velocities shortly after successful RCCA reconstruction. Collateral flow persists longer but decreases rapidly.

Blood Flow Velocity↗
PubMed1992

Cannula-induced vertebral steal in neonates during extracorporeal membrane oxygenation: detection with color Doppler US.

To determine whether flow through the subclavian artery might be affected during extracorporeal membrane oxygenation (ECMO), 40 neonates were examined with color Doppler ultrasound during and after ECMO. Retrograde flow in the right vertebral artery, noted in 12 of the 40 neonates (30%), was consistent with vertebral steal. Brachial systolic velocity was significantly less (P less than .01) on the right than on the left side in neonates both with and without vertebral steal. When the arterial cannula was removed after ECMO, vertebral artery flow became antegrade with symmetric velocity. Brachial velocities became symmetric in infants without vertebral steal, but mild asymmetry persisted in neonates who had had vertebral steal. Only one neonate had clinical signs of arm ischemia, which resolved promptly after removal of the cannula. No surviving neonates (n = 11) had neurologic findings related to the vertebrobasilar insufficiency over a 12-22-month period of observation. Vertebral steal appears to be common during ECMO and is resolved after removal of the cannula.

Basilar Artery↗
PubMed1991

Physiologic rate of carrier-mediated Ca2+ entry matches active extrusion in human erythrocytes.

The intracellular Ca2+ concentration of nearly all cells is kept at submicromolar levels. The magnitudes of transmembrane Ca2+ movement that maintain this steady state in the human red blood cell have long been debated. Although there is agreement that the physiologic extrusion of Ca2+ by the well-characterized Ca2+. ATPase amounts to 45 mumol/liter cells per h (1982. Nature (Lond.). 298:478-481), the reported passive entry rates in physiological saline (2-20 mumol/liter cells per h) are all substantially lower. This discrepancy could be due to incomplete inhibition of the pump in the previous measurements of Ca2+ entry. We therefore examined both rate and mechanism of entry after completely inactivating the pump. This required pretreatment with iodoacetamide (to lower the intracellular ATP concentration) and vanadate (to inhibit any residual Ca2+ pump activity). The rate of Ca2+ entry (53 mumol/liter cells per h) was now found to be comparable to the accepted extrusion rate. Entry closely obeyed Michaelis-Menten kinetics (Vmax = 321 +/- 17 nmol Ca/g dry wt per h, Km = 1.26 +/- 0.13 mM), was competitively inhibited by external Sr2+ (Ki = 10.8 +/- 1.2 mM), and was accelerated by intracellular Ca2+. 45Ca2+ efflux from these pump-inactivated cells was also accelerated by either external Ca2+ or Sr2+. These accelerating effects of divalent cations on the opposite (trans) face of the membrane rule out a simple channel. Substrate-gated channels are also ruled out: cells equilibrated with 45Ca2+ lost the isotope when unlabeled Ca2+ or Sr2+ was added externally. Thus, passive Ca2+ movements occur predominantly by a reversible carrier-mediated mechanism for which Sr2+ is an alternate substrate. The carrier's intrinsic affinity constants for Ca2+ and Sr2+, 1.46 and 0.37 mM-1, respectively, indicate that Ca2+ is the preferred substrate.

Calcium↗
PubMed1990

Right carotid artery ligation in neonates: classification of collateral flow with color Doppler imaging.

Extracorporeal membrane oxygenation (ECMO) is a lifesaving procedure for neonates that involves permanent ligation of the right common carotid artery. To determine the collateral flow patterns that develop after ligation, 58 infants underwent a total of 115 color Doppler imaging studies during (n = 54), within 1 month after (n = 36), or more than 1 month after (n = 25) ECMO. On the basis of the direction of flow in the right internal carotid artery (ICA) proximal and distal to the right posterior communicating artery (PCoA), 85 of the 115 studies were placed in one of three categories: (a) Anterior communicating artery (ACoA) dominance (n = 22), with retrograde flow in the entire right ICA, was most common during ECMO and was never seen more than 1 month after ECMO. (b) PCoA dominance (n = 47), with antegrade right ICA flow distal to the right PCoA but retrograde proximally, was common during all time periods. (c) External carotid artery (ECA) dominance (n = 16), with antegrade flow throughout the right ICA, was noted in 53% of studies more than 1 month after ECMO but in only 9% of earlier studies. Patterns changed from ACoA to PCoA to ECA dominance in 17 infants, but no change in the opposite direction was noted.

Carotid Arteries↗
PubMed1990

Immunoglobulins and allergy mediators in bronchial asthma.

Seventy four young asthmatics were evaluated to establish the role of immunoglobulins and allergy mediators in clinical asthma. They were categorized into Group I (n = 38) or Group II (n = 36) when having mild or severe bronchospasm respectively. In some patients, reversibility of bronchospasm (n = 35), exercise challenge (n = 24) and intradermal allergy testing (n = 24) were also studied. Seventy subjects had raised IgE levels (greater than 100 KU/L) but these did not correlate with the severity of clinical or exercise induced asthma or with reversibility of bronchospasm. An IgE level greater than 1000 KU/L was associated with more number of positive skin tests. Group II subjects had a shorter blood recalcification time (p less than 0.01) and higher levels of IgG, histamine and serotonin (p less than 0.001). It appears that severe asthma is associated with activation of blood coagulation, and release of allergy mediators contributes to the evolution of bronchospasm. Total IgE predicts the utility of skin testing in asthma.

Adult↗
PubMed1978

Phase II trial of a combination of cyclosphamide, vincristine, and methotrexate in advanced colorectal carcinoma.

Twenty patients with advanced colon cancer were treated with a combination of low-dose weekly cyclophosphamide, vincristine, and methotrexate (COM) after demonstrated progression of disease with 5-fluorouracil (5-FU). Four patients (20%) had partial responses with a median survival of 15 months, six (30%) had disease stabilization for 3--16 months (median survival, 8 months), and ten progressed with a median survival of 5.5 months. All four partial responses and five of six stabilizations occurred in patients with a performance status of 0 or 1. COM appeared well-tolerated and at least as effective as alternative secondary chemotherapy following 5-FU failure.

Adult↗