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Biomedical subjects

S Khan

Publications and source records attributed to S Khan.

At least 163 records · Page 9Linked to original sources
PubMed2000

Characterization and outcome of "hard to mobilize"' lymphoma patients undergoing autologous stem cell transplantation.

A "hard to mobilize" patient was defined as one in whom >or= 1x10(6) CD 34+ cells/kg cannot be obtained after two consecutive large volume aphereses. Forty-four consecutive Hodgkin's and non-Hodgkin's lymphoma patients who underwent autologous peripheral blood stem cell (PBSC) transplant treatment between June 1996 and June 1998 were included in this study. Twenty-one patients (48%) met the definition of "hard to mobilize" (Group I). All the rest of the patients (n=23) were the good mobilizers (Group II). The initial mobilization protocol for most patients was 10 microg/kg of G-CSF alone for both groups. For Group I, 7/21 (33%) patients were unable to achieve a minimal dose of >or= 1x10(6) CD34+ cells/kg even after a second mobilization attempt and/or bone marrow (BM) harvest (n=5). Overall, 11/21 (52%) required an additional mobilization and/or BM harvest. Only 3/21 (14%) patients were able to meet the target cell dose of >or= 2.5x10(6) CD34+ cells/kg (median of 4 apheresis). In contrast, 87% of Group II achieved the target dose with a median of 2 aphereses. Predictors of poor mobilization were greater than two prior treatment regimens (p=0.038) and the WBC count (<25,000/microL) on the first day of apheresis (p=0.053). Nineteen patients in Group I and all Group II completed treatment with a median time to engraftment of ANC>500/microl of 12 and 11 days, and platelet >20x10(3)/microl of 31 and 13 days, respectively. Outcome analysis revealed that 6/19 patients in Group I died of relapse within one year from transplant compared with only 2/23 of Group II who died of relapse (p=0.005, log rank test). There were no treatment related deaths in either group. Independent predictive features for "hard to mobilize" patients are a lack of significant increase in WBC count on the first day of apheresis and the number of prior treatment regimens. Poor mobilization appears to predict a worse outcome after autografting for lymphoma patients.

Actuarial Analysis↗
PubMed1999

Response tuning in bacterial chemotaxis.

Chemotaxis of enteric bacteria in spatial gradients toward a source of chemoattractant is accomplished by increases in the length of swimming runs up the gradient. Biochemical components of the intracellular signal pathway have been identified, but mechanisms for achieving the high response sensitivity remain unknown. Binding of attractant ligand to its receptor inactivates a receptor-associated histidine kinase, CheA, which phosphorylates the signal protein CheY. The reduction in phospho-CheY, CheY-P, levels prolongs swimming runs. Here, the stimulus-response relation has been determined by measurement of excitation responses mediated by the Tar receptor to defined concentration jumps of the attractant, aspartate, administered within milliseconds by photolysis of a photolabile precursor. The bacteria responded to <1% changes in Tar occupancy when adapted to aspartate over concentrations spanning three orders of magnitude. Response amplitudes increased approximately logarithmically with stimulus strength, extending responsiveness over a greater stimulus range. The extent and form of this relation indicates that, in contrast to mechanisms for adaptive recovery, excitation signal generation involves amplification based on cooperative interactions. These interactions could entail inactivation of multiple receptor-CheA signaling complexes and/or simultaneous activation of CheY-P dephosphorylation.

Adaptation, Physiological↗
PubMed1999

Comparison of plasma sample purification by manual liquid-liquid extraction, automated 96-well liquid-liquid extraction and automated 96-well solid-phase extraction for analysis by high-performance liquid chromatography with tandem mass spectrometry.

Three extraction procedures were developed for the quantitative determination of a carboxylic acid containing analyte (I) in human plasma by high-performance liquid chromatography (HPLC) with negative ion electrospray tandem mass spectrometry (MS-MS). The first procedure was based on the manual liquid-liquid extraction (LLE) of the acidified plasma samples with methyl tert.-butyl ether. The second procedure was based on the automation of the manual LLE procedure using 96-well collection plates and a robotic liquid handling system. The third approach was based on automated solid-phase extraction (SPE) using 96-well SPE plates and a robotic liquid handling system. A lower limit of quantitation of 50 pg/ml was achieved using all three extraction procedures. The total time required to prepare calibration curve standards, aliquot the standards and plasma samples, and process a total of 96 standards and samples by manual LLE was three-times longer than the time required for 96-well SPE or 96-well LLE (4 h, 50 min vs. 1 h, 43 min). Even more importantly, the time the bioanalyst physically spent on the 96-well LLE or 96-well SPE procedure was only a small fraction of the time spent on the manual LLE procedure (<10 min vs. 4 h, 10 min). It should be noted that the 96-well SPE procedure incorporated the two steps of evaporation of the eluates to dryness and subsequent reconstitution of the dried extract. The total time required for the 96-well SPE could be reduced by 50% if the eluates were injected directly, eliminating the drying and reconstitution steps, which is achievable when sensitivity is less of an issue.

Automation↗
PubMed1999

Effects of acute restraint stress on endogenous adrenomedullin levels.

Adrenomedullin (ADM) is a 52 amino acid peptide, with a potent hypotensive/vasodilatory action. Levels of ADM are significantly elevated in patients with hypertension, and it has been postulated that such stressor-related increases may serve a regulatory or protective function. The current study assessed the effects of acute restraint stress on ADM levels in regions of the brain, plasma and peripheral tissue including heart, lung and the adrenal glands of rats. This stressor, known to stimulate sympathetic activity as well as the hypothalamic-pituitary adrenal (HPA) axis, produced a significant increase in ADM levels in the pituitary gland, plasma and adrenal glands, all of which are key components of the HPA axis. The results suggest a regulatory and/or protective role for ADM in countering HPA activation following a variety of physiological and psychological stressors.

Acute Disease↗
PubMed1999

Maintenance of steroid metabolism and hormone responsiveness in cryopreserved dog, monkey and human hepatocytes.

The efficient and effective use of hepatocytes from larger species and rare human material requires a reliable storage method for cells not needed on the day of preparation. Cryopreservation would seem to be the only viable alternative. In this study the suitability of a published cryopreservation technique on dog, monkey and human hepatocytes has been examined and the cells were tested for functionality directly after thawing and subsequent to culture using steroid metabolism and hormone responsiveness of glycogen phosphorylase a. Monkey and human hepatocytes appear to survive the freezing and thawing process better than dog cells-the latter losing the ability to respond to adrenergic stimuli and their ability to maintain steroid metabolism in culture. Although monkey and human cells do preserve their steroid metabolising capacity after freeze/thawing, there is not the significant increase in enzyme activity seen during culturing freshly isolated cells. It would appear, therefore, that some damage has occurred to the cells during the freeze/thaw process. As previously noted, Williams' medium E is superior to Ham's F-10 in maintaining enzyme activities in culture. It is suggested that cryopreservation is the way forward for the development of stockpiles of viable hepatocytes for biomedical and toxicological research and development but that further modifications to the process are still necessary to optimise the maintenance of liver-specific functions in the thawed cells.

Androstenedione↗
PubMed1999

A multi-laboratory evaluation of cryopreserved monkey hepatocyte functions for use in pharmaco-toxicology.

Ethical, economic and technical reasons hinder regular supply of freshly isolated hepatocytes from higher mammals such as monkey for preclinical evaluation of drugs. Hence, we aimed at developing optimal and reproducible protocols to cryopreserve and thaw parenchymal liver cells from this major toxicological species. Before the routine use of these protocols, we validated them through a multi-laboratory study. Dissociation of the whole animal liver resulted in obtaining 1-5 billion parenchymal cells with a viability of about 86%. An appropriate fraction (around 20%) of the freshly isolated cells was immediately set in primary culture and various hepato-specific tests were performed to examine their metabolic, biochemical and toxicological functions as well as their ultrastructural characteristics. The major part of the hepatocytes was frozen and their functionality checked using the same parameters after thawing. The characterization of fresh and thawed monkey hepatocytes demonstrated the maintenance of various hepato-specific functions. Indeed, cryopreserved hepatocytes were able to survive and to function in culture as well as their fresh counterparts. The ability for synthesis (proteins, ATP, GSH) and conjugation and secretion of biliary acids was preserved after deep freeze storage. A better stability of drug metabolizing activities than in rodent hepatocytes was observed in monkey. After thawing, Phase I and Phase II activities (cytochrome P450, ethoxycoumarin-O-deethylase, aldrin epoxidase, epoxide hydrolase, glutathione transferase, glutathione reductase and glutathione peroxidase) were well preserved. The metabolic patterns of several drugs were qualitatively and quantitatively similar before and after cryopreservation. Lastly, cytotoxicity tests suggested that the freezing/thawing steps did not change cell sensitivity to toxic compounds.

Adenosine Triphosphate↗
PubMed1999

Catabolism of adipose tissue by a tumour-produced lipid-mobilising factor.

Induction of lipolysis in murine white adipocytes by a tumour lipid-mobilising factor (LMF) was associated with stimulation of adenylate cyclase in adipocyte plasma membrane preparations. Induction of lipolysis was attenuated by the adenylate cyclase inhibitor MDL12330A and the protein kinase A inhibitor H8, suggesting that cAMP was the intracellular mediator of induction. The effect of LMF on adenylate cyclase was responsive to GTP, with low concentrations (0.1 microM) causing stimulation and high concentrations (10 microM) causing inhibition, suggesting the involvement of both stimulatory (Gs) and inhibitory (Gi) guanine nucleotide-binding proteins. At a concentration of 10 microM, propranolol noncompetitively reduced the induction of lipolysis by LMF. Thus, lipolysis in white adipose tissue during the process of cancer cachexia is mediated by a tumour factor which stimulates cAMP production, possibly through a beta-adrenergic receptor.

Adenylyl Cyclase Inhibitors↗
PubMed1999

Augmentation of 12-O-tetradecanoyl 13-phorbol acetate-mediated tumor promoting response by the porphyrin photosensitization of 7,12-dimethyl benz[a]anthracene-initiated murine skin: role of in situ generated reactive oxygen species.

Recently, we have shown that sustained ROS generation by prolonged porphyrin-mediated photosensitization in murine skin acts as a stage I and weak complete tumor promoter. Further to this, in the present study, we show that porphyrin photosensitization of DMBA-initiated murine skin results in the augmentation of TPA-mediated tumor promoting response. The photosensitization increased tumor yield to 15 tumors per mouse as compared to 7.5 tumors per mouse in the group treated with TPA alone. Further, 100% tumor incidence in the TPA-treated photosensitized group occurred at week 11 whereas it occurred at week 19 in the TPA alone treated group. Porphyrin photosensitization slightly decreased the latency period of TPA-mediated tumor formation by 1 week. The TPA-mediated ODC induction (1300% of saline-treated control) has been augmented in the photosensitized group (1950%). However, the amount of [3H]thymidine incorporation was not significantly different in the photosensitized TPA-treated and TPA alone-treated groups. Similarly, TPA treatment in photosensitized animals augmented the depletion of cutaneous glutathione and enhancement of lipid peroxidation. These changes were attenuated in butylated hydroxytoluene-pretreated animals. Our results suggest that cutaneous porphyrin photosensitization augments TPA-mediated tumor promotion in murine skin.

9,10-Dimethyl-1,2-benzanthracene↗
PubMed1999

The effects of copper on blood and biochemical parameters of rainbow trout (Oncorhynchus mykiss).

Metals are released into aquatic systems from many sources, often at sublethal concentrations. The effects of sublethal concentrations of metals on fish are not entirely understood. The objective of this study was to determine the hematological and biochemical effects of a range of copper concentrations (6.4, 16.0, 26.9 microg Cu/L) on rainbow trout (Oncorhynchus mykiss) over a prolonged period of time. Trout were exposed to copper, and, at intervals of 3, 7, 14, and 21 days, selected parameters were evaluated. Hemoglobin, hematocrit, plasma glucose, and plasma cortisol levels were elevated in trout exposed to 26.9 microg Cu/L at day 3 and then returned to levels comparable to control fish. Plasma protein and lactate levels were not significantly altered in trout from any copper treatment. Hepatic copper concentration and hepatic metallothionein mRNA expression were consistently elevated in trout exposed to 26.9 microg Cu/L. Both of these parameters stabilized by day 3, with only hepatic copper concentration showing a further increase at day 21. Hepatic copper concentration and hepatic metallothionein mRNA expression appear to be robust indicators of copper exposure. Most blood-based parameters evaluated appear to be associated with a transitory, nonspecific stress response. The return of elevated hematological and biochemical parameters to control levels after 3 days and the stabilization of hepatic metallothionein mRNA expression and copper concentration over a similar time period suggested acclimation to dissolved copper at 26.9 microg/L. Further analysis of the data on blood-based parameters indicated that certain parameters (hemoglobin, hematocrit, plasma glucose, plasma cortisol) may be useful in field monitoring.

Animals↗
PubMed1999

Actual failure rates: A method of assessing tissue valve reoperation rates.

BACKGROUND: Patients older than age 70 with coronary disease undergoing mitral valve replacement have a limited survival rate, suggesting that few of these patients will require reoperation if they receive a tissue mitral valve. However, traditional actuarial analysis of valve durability censors patients who die before valve failure and doesn't reflect the lower frequency of reoperation in patient subgroups with limited survival rates. METHODS: Actual or cumulative reoperation rates represent the rates of reoperation in patients without censoring for death and estimate the risk that the valve will fail before the patient dies. The actual rates of valve rereplacement were determined in 255 recipients of tissue mitral valves, categorizing patients by age and presence of coronary artery disease at 8 years after surgery and compared with standard actuarial estimates of valve durability. RESULTS: Operative mortality rates were 17.5% for patients with coronary artery disease and 6.0% for those without (P <.001). At 8 years, the actual reoperation rates for patients younger than age 70 with and without coronary artery disease were 9.2% (n = 76) and 10% (n = 90), respectively. In patients older than age 70 without coronary artery disease the reoperation rate was 9.4% (n = 32). In contrast, the actual reoperation rate was only 1.7% (n = 58) in patients older than age 70 with coronary artery disease. This rate was significantly lower (P =.05) than the other groups. The difference in reoperation rates was not significant if standard actuarial analysis was used. CONCLUSIONS: Actual reoperation rates are significantly lower in patients older than 70 years with CAD receiving mitral tissue valves than in younger patients or in patients without coronary artery disease. Calculation of actual reoperation rates provides a unique and clinically useful perspective in evaluating data on prosthetic valve reoperation rates.

Age Factors↗
PubMed1999

Identifying patients at low risk of death from cardiac failure after operation for postinfarct ventricular tachycardia.

BACKGROUND: In unselected patients, cardiac failure accounted for most deaths after antiarrhythmic operation (ER) for postinfarction ventricular tachycardia (VT). This study aimed to determine whether patients at low risk of this outcome could be predicted from a retrospective analysis of variables from 100 consecutive ER patients. METHODS: Thirteen variables suggested by other researchers as predictive of outcome were analyzed. At the time of study, ER was the only therapy available for drug refractory VT. RESULTS: Only emergency ER, wall motion score less than 3 and Killip classification were significantly related to death from cardiac failure. The lack of correlation between emergency ER and variables of ER timing, VT less than 24 hours of ER or VT type implies that the need for emergency ER is also related to ventricular dysfunction. Multivariate analysis identified a group at particularly low risk of death with a specificity of 95%. CONCLUSIONS: Patients at low risk of death after ER can be identified prospectively. In the implantable cardioverter defibrillator era, elective ER is best reserved for such patients. Emergency ER may still be justified in younger patients without comorbidity who will die of VT without it.

Adolescent↗
PubMed1999

Chemotactic responses of Escherichia coli to small jumps of photoreleased L-aspartate.

Computer-assisted motion analysis coupled to flash photolysis of caged chemoeffectors provides a means for time-resolved analysis of bacterial chemotaxis. Escherichia coli taxis toward the amino acid attractant L-aspartate is mediated by the Tar receptor. The physiology of this response, as well as Tar structure and biochemistry, has been studied extensively. The beta-2, 6-dinitrobenzyl ester of L-aspartic acid and the 1-(2-nitrophenyl)ethyl ether of 8-hydroxypyrene-1,3,6-tris-sulfonic acid were synthesized. These compounds liberated L-aspartate and the fluorophore 8-hydroxypyrene 1,3,6-tris-sulfonic acid (pyranine) upon irradiation with near-UV light. Photorelease of the fluorophore was used to define the amplitude and temporal stability of the aspartate jumps employed in chemotaxis experiments. The dependence of chemotactic adaptation times on aspartate concentration, determined in mixing experiments, was best fit by two Tar aspartate-binding sites. Signal processing (excitation) times, amplitudes, and adaptive recovery of responses elicited by aspartate jumps producing less than 20% change in receptor occupancy were characterized in photorelease assays. Aspartate concentration jumps in the nanomolar range elicited measurable responses. The response threshold and sensitivity of swimming bacteria matched those of bacteria tethered to glass by a single flagellum. Stimuli of similar magnitude, delivered either by rapid mixing or photorelease, evoked responses of similar strength, as assessed by recovery time measurements. These times remained proportional to change in receptor occupancy close to threshold, irrespective of prior occupancy. Motor excitation responses decayed exponentially with time. Rates of excitation responses near threshold ranged from 2 to 7 s-1. These values are consistent with control of excitation signaling by decay of phosphorylated pools of the response regulator protein, CheY. Excitation response rates increased slightly with stimulus size up to values limited by the instrumentation; the most rapid was measured to be 16 +/- 3 (SE) s-1. This increase may reflect simultaneous activation of CheY dephosphorylation, together with inhibition of its phosphorylation.

Adaptation, Physiological↗
PubMed1999

Role of the cellular redox state in modulating acute ethanol toxicity in isolated hepatocytes.

OBJECTIVES: To propose a mechanism for ethanol induced hepatocytotoxicity. DESIGN AND METHODS: Hepatocytotoxicity was determined at various concentrations of oxygen and agents involved in NADH metabolism. RESULTS: At 1% O2, hepatocytes were nearly 8-fold more susceptible to ethanol than at 95% O2 (carbogen). Cytotoxicity at 1% O2 was enhanced in the presence of glycolytic substrates that generate NADH (e.g., sorbitol or xylitol), and prevented by glycolytic substrates that reoxidise NADH (e.g., fructose or dihydroxyacetone). Susceptibility to ethanol correlated with the cytosolic redox state (lactate; pyruvate ratio). Cytotoxicity also correlated with reactive oxygen species (ROS) formation. Cytotoxicity was averted by ROS scavengers or the ferric chelator desferoxamine but was increased by hydroxylamine, a catalase inhibitor, or by prior glutathione depletion. Ethanol induced cytotoxicity was also decreased by inhibitors of alcohol/aldehyde dehydrogenases or CYP2E1, an alcohol inducible cytochrome P450. CONCLUSIONS: A cytotoxic mechanism was proposed where the sustained increase in NADH levels, resulting from ethanol metabolism, maintains CYP2E1 in a more reduced state that increases ROS formation.

Alcohol Dehydrogenase↗
PubMed1999

Fulminant septicaemic syndrome of Bacillus cereus: three case reports.

Three patients with acute leukaemia, who were severely neutropenic and iatrogenically immunosuppressed post-chemotherapy, developed rapidly fatal septicaemic shock and coma caused by Bacillus cereus (B. cereus). The illness was marked by two phases: a mild febrile illness lasting 6-14 h and accompanied by subtle symptoms of autonomic sympathetic nervous system overactivity, and a second short fulminant one, marked by high fever of 40-41 degrees C accompanied by major central nervous system disturbances, and ending with deep coma and brain stem dysfunction. One patient developed the sepsis in spite of 4 days of coverage with amikacin. In the other two patients, amikacin was commenced at the earliest phase of the infection, but failed to influence the outcome. This form of B. cereus sepsis in neutropenic patients seems to be caused by strains capable of causing bacteraemia and meningitis and has the ability to produce a substance that causes leptomeningeal and neuronal necrosis. Lack of early clinical and laboratory markers inevitably leads to death. Use of antibiotics effective against B. cereus and capable of achieving high concentrations in the cerebrospinal fluid. and identification and neutralization of the necrotizing substance may hopefully help to reverse this fatal illness.

Acute Disease↗
PubMed1999

Gastroesophageal reflux disease in children.

In the pediatric population, gastroesophageal reflux most often presents in infancy as effortless regurgitation, but pathologic GERD is accompanied by signs of malnutrition, respiratory diseases, and esophagitis or its complications. Because of the distinctive pathophysiology predisposing infants to GERD, the diagnostic approach must begin with a thorough history that determines the extent of further diagnostic tests and the course of management. Empiric therapy assumes importance in infants with GERD because of the limited differential diagnoses in consideration. Conservative therapy is of utmost importance because of the unique provocative factors in the pathophysiology of infantile GERD. Prokinetic pharmacotherapy takes precedence over acid suppression because of the more important role of motility factors compared with acid secretion in infantile GERD.

Child↗
PubMed1999

Development of human papillomavirus plasmids capable of episomal replication in human cell lines.

Gene delivery into human cells has been facilitated by the development of viral vector systems. These vectors have shown great potential for the efficient delivery of therapeutic genes into human cells. A problem with many of the existing systems, however, is the integration of these vectors into the chromosome which affects the length of gene expression and may promote oncogenic transformation. In an effort to develop viral vectors that can replicate extrachromosomally in human cells, we have generated human papillomavirus (HPV) plasmids containing all the elements required for replication on a single DNA molecule. HPV plasmids containing the viral E1 and E2 genes (or the E1 gene alone) and an origin of replication were shown to replicate to significant levels in the transfected human cervical carcinoma C-33A cell line. Since approaches towards the possible gene therapy of cystic fibrosis (CF) are currently under intensive investigation, we have also tested short-term replication of HPV plasmids in the IB3 cell line derived from a CF patient. Our results demonstrate that HPV plasmids are capable of extrachromosomal replication in these cell lines and may potentially be important vectors for the delivery of therapeutic genes into human cells.

Cystic Fibrosis↗
PubMed1999

2'-carboxy-D-arabitinol 1-phosphate protects ribulose 1, 5-bisphosphate carboxylase/oxygenase against proteolytic breakdown.

Trypsin-catalysed cleavage of purified ribulose 1,5-bisphosphate carboxylase/oxygenase (Rubisco) and the resultant irreversible loss of carboxylase activity were prevented by prior incubation with the naturally occurring nocturnal Rubisco inhibitor 2'-carboxy-D-arabitinol 1-phosphate (CA1P), as well as with ribulose 1,5-bisphosphate (RuBP), Mg2+ and CO2. CA1P also protected Rubisco from loss of activity caused by carboxypeptidase A. When similar experiments were carried out using soluble chloroplast proteases, CA1P was again able to protect Rubisco against proteolytic degradation and the consequent irreversible loss of catalytic activity. Thus, CA1P prevents the proteolytic breakdown of Rubisco by endogenous and exogenous proteases. In this way, CA1P may affect the amounts of Rubisco protein available for photosynthetic CO2 assimilation. Rubisco turnover (in the presence of RuBP, Mg2+ and CO2) may confer similar protection against proteases in the light.

Carboxypeptidases↗