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Biomedical subjects

Shahid Khan

Publications and source records attributed to Shahid Khan.

11 recordsLinked to original sources

Differential activation of Escherichia coli chemoreceptors by blue-light stimuli.

Enteric bacteria tumble, swim slowly, and are then paralyzed upon exposure to 390- to 530-nm light. Here, we analyze this complex response in Escherichia coli using standard fluorescence microscope optics for excitation at 440 +/- 5 nm. The slow swimming and paralysis occurred only in dye-containing growth media or buffers. Excitation elicited complete paralysis within a second in 1 muM proflavine dye, implying specific motor damage, but prolonged tumbling in buffer alone. The tumbling half-response times were subsecond for onset but more than a minute for recovery. The response required the chemotaxis signal protein CheY and receptor-dependent activation of its kinase CheA. The study of deletion mutants revealed a specific requirement for either the aerotaxis receptor Aer or the chemoreceptor Tar but not the Tar homolog Tsr. The action spectrum of the wild-type response was consistent with a flavin, but the chromophores remain to be identified. The motile response processed via Aer was sustained, with recovery to either step-up or -down taking more than a minute. The response processed via Tar was transient, recovering on second time scales comparable to chemotactic responses. The response duration and amplitude were dependent on relative expression of Aer, Tar, and Tsr. The main response features were reproduced when each receptor was expressed singly from a plasmid in a receptorless host strain. However, time-resolved motion analysis revealed subtle kinetic differences that reflect the role of receptor cluster interactions in kinase activation-deactivation dynamics.

Bacterial Proteins↗

Dexamethasone as antiemetic during gynaecological laparoscopic surgery.

OBJECTIVE: To evaluate the efficacy of intravenous dexamethasone for preventing postoperative nausea and vomiting in women undergoing gynecological laparoscopic surgery. DESIGN: Double blind trial. PLACE AND DURATION OF STUDY: December 2001 to June 2002 at Hayatabad Medical Complex, Peshawar. PATIENTS AND METHODS: A 100 admitted female patients of ASA-1-II scheduled for diagnostic gynecological laparoscopic surgery were included in this study. Patients with severe systemic or endocrine disease who had predisposing factors for delayed gastric emptying, such as diabetes, chronic cholecystitis or neuromuscular disorders were excluded. In addition patients who suffered from postoperative nausea and vomiting (PONV) or had received an antiemetic agent or narcotic medications within last 24 hours were also excluded. Patients were divided into two equal groups, patients in one group were given dexamethasone while saline was injected to patients in the second group. Nausea and vomiting were assessed immediately after operation, at 1 hour interval for 4 hours in the recovery and from 4-10 hours in the Ward. Result was compared in two groups by chi-square test. RESULTS: During patient s stay in the Postanaesthesia Care Unit (4 hours postoperatively) 26% patients in the dexamethasone group in comparison with 54% of patients in the saline group reported PONV (P < 0.01). Sixteen percent of patients in the dexamethasone group, in comparison with 28% of patients in the saline group needed rescue antiemtic (P 0.05), postoperative nausea vomiting (P < 0.01). During the postoperative observation period of 10 hours, 42% of the patients in the dexamethasone group in comparison with 82% of patients in the saline group reported postoperative nausea and vomiting (P < 0.001). CONCLUSION: Dexamethasone significantly reduced the incidence of postoperative nausea and vomiting in women undergoing laparoscopic surgery. Furthermore, it is freely available, is less costly and has few side-effects. So, it should be more frequently used as prophylactic antiemetic in women undergoing gynaecological laparoscopic surgery.

Adult↗

Novel strains of Salmonella typhimurium as potential vectors for gene delivery.

DNA vaccines are known to induce long-term antigen specific cellular responses. We tested two new strains of Salmonella typhimurium, one carrying a mutation in a SPI-2 gene and the aroC-gene and another carrying mutations in the sifA- and aroC-genes, as potential DNA vaccine delivery vehicles. We compared them with the SL7207 strain and found that the new strains were more invasive, and that they were efficient mediators of gene transfer in vitro using EGFP as reporter gene. We tested the ability of the new strains to survive within the spleen, liver and mesenteric lymph nodes and evaluated their safety in C57/BL/6J mice.

Animals↗

Biphasic excitation by leucine in Escherichia coli chemotaxis.

Leucine concentration jumps (applied by photolysis of inert derivatives) triggered swim or tumble responses in Escherichia coli mutants lacking Tsr or Tar, respectively. Wild-type E. coli bacteria were attracted in spatial assays when the initial leucine concentration difference was 5 to 120 micro M but were repulsed when it was over 0.5 mM. Their responses to concentration jumps confirmed earlier deductions regarding biphasic excitation.

Chemotaxis↗

The fast tumble signal in bacterial chemotaxis.

We have analyzed repellent signal processing in Escherichia coli by flash photorelease of leucine from photolabile precursors. We found that 1). response amplitudes of free-swimming cell populations increased with leucine jump concentration, with an apparent Hill coefficient of 1.3 and a half-maximal dose of 14.4 microM; 2). at a 0-0.5 mM leucine concentration jump sufficient to obtain a saturation motile response, the swimming cell response time of approximately 0.05 s was several-fold more rapid than the motor response time of 0.39 +/- 0.18 s measured by following the rotation of cells tethered by a single flagellum to quartz coverslips; and 3). the motor response time of individual cells was correlated with rotation bias but not cell size. These results provide information on amplification, rate-limiting step, and flagellar bundle mechanics during repellent signal processing. The difference between the half-maximal dose for the excitation response and the corresponding value reported for adaptation provides an estimate of the increase in the rate of formation of CheYP, the phosphorylated form of the signal protein CheY. The estimated increase gives a lower limit receptor kinase coupling ratio of 6.0. The magnitude and form of the motor response time distribution argue for it being determined by the poststimulus switching probability rather than CheYP turnover, diffusion, or binding. The temporal difference between the tethered and swimming cell response times to repellents can be quantitatively accounted for and suggests that one flagellum is sufficient to cause a measurable change of direction in which a bacterium swims.

Bacterial Proteins↗

Binding of the chemotaxis response regulator CheY to the isolated, intact switch complex of the bacterial flagellar motor: lack of cooperativity.

In bacteria, the chemotactic signal is greatly amplified between the chemotaxis receptors and the flagellar motor. In Escherichia coli, part of this amplification occurs at the flagellar switch. However, it is not known whether the amplification results from cooperativity of CheY binding to the switch or from a post-binding step. To address this question, we purified the intact switch complex (constituting the switch proteins FliG, FliM, and FliN and the scaffolding protein FliF) in quantities sufficient for biochemical work and used it to investigate whether the binding of CheY to the switch complex is cooperative. As a negative control, we used complexes of switchless basal bodies, formed from the proteins FliF and FliG and similarly isolated. Using double-labeling centrifugation assays for binding, we found that CheY binds to the isolated, intact switch complex in a phosphorylation-dependent manner. We observed no significant phosphorylation-dependent binding to the negative control of the switchless basal body. The dissociation constant for the binding between the switch complex and phosphorylated CheY (CheY approximately P) was 4.0 +/- 1.1 microm, well in line with the published range of CheY approximately P concentrations to which the flagellar motor is responsive. Furthermore, the binding was not cooperative (Hill coefficient approximately 1). This lack of CheY approximately P-switch complex binding cooperativity, taken together with earlier in vivo studies suggesting that the dependence of the rotational state of the motor on the fraction of occupied sites at the switch is sigmoidal and very steep (Bren, A., and Eisenbach, M. (2001) J. Mol. Biol. 312, 699-709), indicates that the chemotactic signal is amplified within the switch, subsequent to the CheY approximately P binding.

Bacterial Physiological Phenomena↗

Variable symmetry in Salmonella typhimurium flagellar motors.

Electron cryomicroscopy of rotor complexes of the Salmonella typhimurium flagellar motor, overproduced in a nonmotile Escherichia coli host, has revealed a variation in subunit symmetry of the cytoplasmic ring (C ring) module. C rings with subunit symmetries ranging from 31 to 38 were found. They formed a Gaussian distribution around a mean between 34 and 35, a similar number to that determined for native C rings. C-ring diameter scaled with the number of subunits, indicating that the elliptical-shaped subunits maintained constant intersubunit spacing. Taken together with evidence that the M ring does not correspondingly increase in size, this finding indicates that rotor assembly does not require strict stoichiometric interactions between the M- and C-ring subunits. Implications for motor function are discussed.

Bacterial Proteins↗

Two msbB genes encoding maximal acylation of lipid A are required for invasive Shigella flexneri to mediate inflammatory rupture and destruction of the intestinal epithelium.

Shigella flexneri is a Gram-negative pathogen that invades and causes inflammatory destruction of the human colonic epithelium, thus leading to bloody diarrhea and dysentery. A type III secretion system that delivers effector proteins into target eukaryotic cells is largely responsible for cell and tissue invasion. However, the respective role of this invasive phenotype and of lipid A, the endotoxin of the Shigella LPS, in eliciting the inflammatory cascade that leads to rupture and destruction of the epithelial barrier, was unknown. We investigated whether genetic detoxification of lipid A would cause significant alteration in pathogenicity. We showed that S. flexneri has two functional msbB genes, one carried by the chromosome (msbB1) and the other by the virulence plasmid (msbB2), the products of which act in complement to produce full acyl-oxy-acylation of the myristate at the 3' position of the lipid A glucosamine disaccharide. A mutant in which both the msbB1 and msbB2 genes have been inactivated was impaired in its capacity to cause TNF-alpha production by human monocytes and to cause rupture and inflammatory destruction of the epithelial barrier in the rabbit ligated intestinal loop model of shigellosis, indicating that lipid A plays a significant role in aggravating inflammation that eventually destroys the intestinal barrier. In addition, neutralization of TNF-alpha during invasion by the wild-type strain strongly impaired its ability to cause rupture and inflammatory destruction of the epithelial lining, thus indicating that TNF-alpha is a major effector of epithelial destruction by Shigella.

Acylation↗

The early clinical and angiographic outcome of sequential coronary artery bypass grafting with the off-pump technique.

OBJECTIVE: The emergence of the off-pump coronary artery bypass technique has made surgeons consider combining it with other techniques developed and learned with conventional coronary artery bypass grafting with cardiopulmonary bypass. One of these techniques is the construction of a sequential graft to bypass more than one coronary vessel. The purpose of this study is to review the outcome of combining sequential coronary artery bypass grafting with off-pump techniques. METHODS: We retrospectively reviewed the records of 45 consecutive patients who underwent isolated coronary bypass surgery with off-pump and sequential grafting techniques at Harefield Hospital (Harefield, UK) between July 1999 and December 2000. The registry database, medical notes and charts were studied for preoperative and postoperative data of the patients. Ten patients consented and underwent early postoperative angiography to check the quality of the grafts and anastomoses. RESULTS: There were no deaths among the study patients. Morbidity consisted of atrial fibrillation in 6 patients (13.3%), leg wound infection in 2 patients (4.4%), and pleural effusion in 1 patient (2.2%). Early angiography of the 10 consenting patients revealed 10 patent sequential grafts (100%) with 20 satisfactory end-to-side and side-to-side anastomoses (100%). CONCLUSION: The combination of sequential grafting and off-pump techniques is feasible, is safe, and provides good early clinical and angiographic outcomes.

Adult↗

Multivessel off-pump coronary artery bypass grafting can be taught to trainee surgeons.

BACKGROUND: Although Off-Pump Coronary Artery Bypass (OPCAB) surgery is being increasingly explored and practised in many cardiac units worldwide, there have been only few reports documenting the training of surgeons in this new technique. The purpose of this study was to address the reproducibility of the OPCAB in a unit where this technique is used extensively. METHODS: Registry data, notes, and charts of 64 patients who were operated on by four trainee cardiac surgeons over a period of thirteen months at Harefield Hospital, were reviewed retrospectively. These trainees were part of an accredited training program for cardiothoracic training and were trained by a single consultant trainer in a cardiac unit after it had an established recent experience in performing nonselective OPCAB for all in-coming patients. Five (7.8%) patients (with 17 distal anastomoses) consented and underwent early postoperative angiography to check the quality of the grafts and anastomoses. RESULTS: The mean age of the study patients was 65.6 and the mean Parsonnet score was 9.4. There was a mean of 2.9 grafts per patient and circumflex territory anastomoses were performed in 48 (75%) patients. No operation required conversion to Cardiopulmonary Bypass (CPB). Angiography of the five patients revealed 17 satisfactory (100%) distal anastomoses. CONCLUSION: With appropriate training, it is possible for trainees to learn OPCAB and perform multivessel revascularization in relatively high-risk patients with good results.

Adult↗