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Biomedical subjects

M R Williams

Publications and source records attributed to M R Williams.

At least 19 recordsLinked to original sources

Recurrent painful unilateral gynaecomastia-interactions between hyperthyroidism and hypogonadism.

We report an unusual case of recurrent, painful, unilateral gynaecomastia (GM) in an elderly male with relapsing Graves' hyperthyroidism and co-existing primary hypogonadism. This patient presented to the Breast Clinic with a 4-month history of painful, right GM. Malignancy was excluded but T3 was noted to be raised at 7.3 pmol l(-1) (normal 3.5-5.5) with a suppressed thyroid-stimulating hormone. Testosterone, luteinizing hormone and follicle-stimulating hormone were consistent with primary hypogonadism. He was later referred to physicians with night sweats and painful right GM. FT3 was 7.4 and carbimazole was commenced. Within 4 months, the night sweats and right GM had resolved but he became hypothyroid. When carbimazole was stopped, right GM recurred together with hyperthyroidism. The male breast, which is sensitive to subtle changes in T/E2 ratio, is more likely to be stimulated in an elderly male with hyperthyroidism and pre-existing hypogonadism, and hence recurrence of GM with relapsing hyperthyroidism. Recognition of this association is clinically relevant to avoid unnecessary investigations and undue patient anxiety, and to facilitate appropriate early diagnosis and treatment.

Aged, 80 and over↗

Skimboarding: a new danger in the surf?

Skimboarding is an increasingly popular activity on the beach. Over a six week period we saw 10 patients with fractures or dislocations, half of whom required operative intervention. The majority (80%) were of the lower limb, some of which were severe.

Adolescent↗

Enzymatic bioremediation: from enzyme discovery to applications.

1. Enzymatic bioremediation is potentially a rapid method of removing environmental pesticide residues. Applications include the treatment of residues resulting from agricultural production and processing industries, such as the treatment of irrigation waters, surface-contaminated fruit and vegetables and spent dip liquors. 2. A specific application for some organophosphate-degrading enzymes involves detoxification of nerve agent stockpiles. Effective and affordable remediation requires highly specialized enzymes, so protein engineering techniques are being used to improve properties of various source enzymes to enhance catalytic rates, stability and substrate range. 3. Trials with an optimized organophosphate-degrading enzyme have shown the feasibility of such technology in various applications. 4. The enzymes developed for environmental remediation for specific pesticide classes also have applications as antidotes for high-dose pesticide poisonings and as prophylaxis for people at risk of high pesticide doses.

Animals↗

Burn injuries after latissimus dorsi breast reconstruction in a cold climate.

The transposed and 'native' skin of a reconstructed breast using autogenous tissue transfer has impaired sensation. Two cases of thermal injury following latissimus dorsi breast reconstruction are reported. Both cases were due to the overnight application of a hot-water bottle to the reconstructed breast. We suggest that all patients should be warned of this potential complication especially in cold climates.

Journal Article↗

Replication-independent assembly of an insect virus (Tetraviridae) in plant cells.

Infectious virions of the insect RNA virus Helicoverpa armigera stunt virus (HaSV; Omegatetravirus, Tetraviridae) were assembled in cultured plant protoplasts of Nicotiana plumbaginifolia in the absence of detectable replication. Assembly of the virus, which has not been grown in cell culture, required cotransfection of a DNA plasmid expressing the HaSV capsid gene in combination with either genomic RNA or with DNA plasmids carrying the complete cDNAs to the two HaSV genomic RNAs. Each cDNA was placed under the control of the cauliflower mosaic virus 35S promoter and followed by a cis-acting ribozyme so that the resultant transcripts corresponded precisely to the two genomic RNAs. Protoplast assembly of infectious particles was confirmed by EM and bioassay of host insect larvae, which became diseased and produced virus particles confirmed as HaSV. Variant transcripts carrying nonviral sequences at either or both termini of the RNAs showed no infectivity, except for RNA2 carrying only a 3' terminal extension. No replication of HaSV in protoplasts was detected in pulse-labeling and blotting experiments. Insects showed less severe disease symptoms when fed protoplasts transfected with only the RNA1 and coat protein plasmids. The symptomatic larvae contained only RNA1 and failed to yield infectious progeny virus, suggesting that RNA1 is capable of self-replication. This novel plasmid-based system confirms that the reported sequence of HaSV represents an infective genome and establishes a procedure for the reverse genetics of a tetravirus.

Animals↗

Phosphorylation of the protein kinase mutated in Peutz-Jeghers cancer syndrome, LKB1/STK11, at Ser431 by p90(RSK) and cAMP-dependent protein kinase, but not its farnesylation at Cys(433), is essential for LKB1 to suppress cell vrowth.

Peutz-Jeghers syndrome is an inherited cancer syndrome that results in a greatly increased risk of developing tumors in those affected. The causative gene is a protein kinase termed LKB1, predicted to function as a tumor suppressor. The mechanism by which LKB1 is regulated in cells is not known. Here, we demonstrate that stimulation of Rat-2 or embryonic stem cells with activators of ERK1/2 or of cAMP-dependent protein kinase induced phosphorylation of endogenously expressed LKB1 at Ser(431). We present pharmacological and genetic evidence that p90(RSK) mediated this phosphorylation in response to agonists that activate ERK1/2 and that cAMP-dependent protein kinase mediated this phosphorylation in response to agonists that activate adenylate cyclase. Ser(431) of LKB1 lies adjacent to a putative prenylation motif, and we demonstrate that full-length LKB1 expressed in 293 cells was prenylated by addition of a farnesyl group to Cys(433). Our data suggest that phosphorylation of LKB1 at Ser(431) does not affect farnesylation and that farnesylation does not affect phosphorylation at Ser(431). Phosphorylation of LKB1 at Ser(431) did not alter the activity of LKB1 to phosphorylate itself or the tumor suppressor protein p53 or alter the amount of LKB1 associated with cell membranes. The reintroduction of wild-type LKB1 into a cancer cell line that lacks LKB1 suppressed growth, but mutants of LKB1 in which Ser(431) was mutated to Ala to prevent phosphorylation of LKB1 were ineffective in inhibiting growth. In contrast, a mutant of LKB1 that cannot be prenylated was still able to suppress the growth of cells.

AMP-Activated Protein Kinase Kinases↗

Direct cardiac compression for cardiogenic shock with the CardioSupport system.

Epicardial direct cardiac compression for cardiogenic shock avoids a blood surface interface with associated thromboembolic and immunologic sequelae and could be placed rapidly with technical ease. The Cardio Technologies device provides synchronized biventricular cardiac compression, is placed via a thoracotomy, and remains on the heart without need for sutures. In preclinical work, the system has successfully restored cardiac function to near normal in the setting of heart failure. The CardioSupport system offers an attractive and novel alternative for treating cardiogenic shock and is being prepared for upcoming clinical trials.

Animals↗

Indications and patient selection for mechanical ventricular assistance.

Cardiac assist devices have become an important component of transplantation programs as they successfully bridge unsalvageable patients who would otherwise die. The indications for a device can still be classified into short-term and long-term situations. The short-term indications have expanded into areas such as postcardiotomy failure, high-risk cardiac operations, and acute myocardial infarction with results that were not previously possible in the absence of a well-established mechanical assistance program. Appropriate patient selection remains challenging and perhaps the most important attribute of a successful ventricular assist program. Although few exact criteria can define patients who are not eligible, several considerations and screening scales can help determine a particular patient's suitability. Specific attention must be given to right heart function, neurologic status, existing infections, renal function, and transplantation suitability. The future of this field will not only be in technological advances with devices but in optimization of patient selection and expanding indications such as permanent replacement therapy.

Algorithms↗

Surgical treatment of atrial fibrillation using radiofrequency energy.

BACKGROUND: The Maze III procedure for atrial fibrillation (AF) is effective but has not been used widely due to its complexity, bleeding risk, and added operative time. Surgical radiofrequency ablation may simplify the procedure and make intraoperative correction of AF more accessible and widely performed. METHODS: Endocardial pulmonary venous isolation was performed on 48 patients with AF undergoing concurrent operation using temperature-controlled radiofrequency energy delivered through a hand-held flexible probe. Additional right-sided lesions were made at the surgeon's discretion. RESULTS: Forty-two patients were appropriate for analysis (6 died). These patients had an AF duration of 4.8 +/- 6.4 years. At a mean follow-up of 138 +/- 96 days, 34 patients were in sinus rhythm. We were unable to demonstrate a difference in outcome based on AF duration, left atrial size, or addition of right-sided lesions. CONCLUSIONS: Radiofrequency atrial ablation was effective in 81% of patients with AF at restoring sinus rhythm at an average follow-up of 4 months. This procedure is simple to perform and should broaden the number of patients that receive an AF treatment procedure during concurrent cardiac operation.

Adult↗

Variations in endothelial function and arterial compliance during the menstrual cycle.

Female sex hormones have been implicated in the cardioprotection of premenopausal women. However, the cardiovascular actions of these hormones and the effects of their natural fluctuations during the menstrual cycle are not fully understood. We studied changes in vascular function during the menstrual cycle in 15 healthy premenopausal women. Four noninvasive procedures were performed during the early follicular (EF), late follicular (LF), early luteal (EL), and late luteal (LL) phases: flow-mediated dilatation (FMD) of the brachial artery during reactive hyperemia, laser Doppler velocimetry (LDV) with direct current iontophoresis of acetylcholine (ACh) and nitroprusside, whole body arterial compliance (WBAC), and pulse wave velocity. Hormone levels were consistent with predicted cycle phase and showed that all subjects ovulated during the cycle studied. FMD, LDV with ACh, and WBAC varied cyclically, with significant increases from the F to LF phase, sharp falls in the EL phase, and significant recoveries in the LL phase. These changes were most marked for FMD [EF, 8.8 +/- 0.6% (mean +/- SEM); LF, 10.0 +/- 0.7; EL, 4.2 +/- 0.6; LL, 8.6 +/- 0.9] and the LDV response to ACh (EF, 2.7 +/- 0.2 V/min; LF, 3.3 +/- 0.4; EL, 1.8 +/- 0.3; LL, 2.7 +/- 0.4). WBAC changed similarly (EF, 0.58 +/- 0.08 arbitrary units; LF, 0.84 +/- 0.06; EL, 0.65 +/- 0.05; LL, 0.68 +/- 0.06). Sodium nitroprusside-induced vasodilatation decreased significantly from EF to EL, with no other significant difference, and pulse wave velocity did not vary significantly over the four time points. Conductance and resistance artery endothelial reactivity and smooth muscle sensitivity to nitric oxide and arterial compliance are modulated significantly in response to the changing hormonal patterns of the menstrual cycle. These findings emphasize the importance of menstrual phase in the interpretation of data on endothelial function and may provide insights into the mechanisms underlying sex differences in cardiovascular risk and other disease processes in premenopausal women.

Adult↗

Response of cotton to prebloom square loss.

In 1996 and 1997, various intensities of prebloom square removal were applied to three cultivars of cotton grown in Mississippi. With the exception of one cultivar in 1997, all cultivars were B. thuringiensis (Bt)-transgenic cotton. At harvest, the number of bolls and seed cotton weight was recorded for all plants in each square removal treatment. All cultivars responded similarly to square loss. A yield increase (overcompensation) was observed in the treatment where all squares were removed from the plant one week after squaring began. Only the treatment where all squares were removed before bloom significantly reduced yield and caused a large (>7 d) delay in crop maturation. Otherwise, moderate levels of square removal (approximately 20-50% of prebloom squares) had little impact on overall lint production. However, the patterns of cotton production on the plants were significantly influenced by the square removal treatments. The removal of relatively more or larger squares increased seed cotton production in late-season fruiting cohorts and on 'vegetative' branches. Compensation for square loss occurred by increasing the relative number and weight of bolls produced subsequent to early-season square removal. Typically, early-season square loss increased the value of later-season fruiting cohorts, especially the midseason cohorts and bolls on vegetative branches. The implications of prebloom square loss, including the compensatory ability of the cotton plant, on insect management are discussed.

Animals↗

Role of the endoplasmic reticulum in shaping calcium dynamics in human lens cells.

PURPOSE: Localized cortical cataracts in the human lens have been shown to involve a selective increase in calcium with no change in sodium content. Recent studies in the rat lens in vitro have shown that the store-operated channel is highly selective for calcium over sodium, and therefore this channel was characterized further in human lens cells. METHODS: Human primary cultures were initiated from epithelial explants and passaged onto coverslips. After incorporating Fura-2, agonist- or thapsigargin-induced changes in cytosolic calcium were monitored and calibrated using fluorometric digital imaging techniques. RESULTS: Histamine and adenosine triphosphate (ATP; 10 microM) induced a large transient increase in cytosolic calcium followed by a maintained lower plateau phase in the continued presence of the calcium-signaling agonist. The second phase was abolished by removing external calcium and represented the contribution from the store-operated influx. The store-operated pathway was blocked by inorganic agents such as zinc and nickel (100 microM) but was insensitive to the voltage-sensitive calcium channel blocker, nifedipine (1 mM). Depolarizing the membrane voltage by raising the external potassium (75 mM) also blocked the influx. Similar results were obtained if the store was first emptied directly using thapsigargin (1 microM), and with this agent it was also possible to observe the very slow activation and inactivation kinetics (>10 seconds) of the channel. Addition of manganese to the bathing medium initiated a quench of Fura-2 isobestic fluorescence that was enhanced 2.9 +/- 0.3-fold after 10 microM ATP addition. There was a delay of 82 +/- 16 seconds between initiation of the calcium spike and the Mn2+ quench rate, indicating the presence of a delayed entry pathway. In the resting state, removal of, or increasing extracellular calcium concentration 10-fold did not perturb the level of cytosolic Ca2+. Similar maneuvers performed after agonist- or thapsigargin-induced store depletion of intracellular stores brought about dramatic changes in cytosolic Ca2+ consistent with the activation of a Ca2+ entry pathway. Lower concentrations of agonist induced oscillations of Ca2+ that continued for a short time in Ca-free solution. No increase in Mn2+ quench rate was associated with oscillations. A 100-microM zinc- and KCl-induced blockade of Ca2+ entry had no effect on the form of agonist-induced oscillations. Inhibition of Ca2+ influx by zinc (100 microM) converted a sustained Ca2+ response to a train of repetitive Ca2+ spikes. CONCLUSIONS: Human lens cells normally have very low Ca2+ permeability. Depletion of intracellular stores by agonists or thapsigargin initiates a Ca2+ entry pathway that is not required for the Ca2+ oscillations induced by low concentrations of agonist. This potentially provides a signal transduction mechanism with minimal risk of Ca2+ overload to the lens, whereas overactivation of the store-operated channel is a possible way of increasing calcium in the lens and could explain the distribution found in localized cataracts.

Adenosine Triphosphate↗

A comparison of consecutive off-pump versus conventional coronary artery bypass.

BACKGROUND: Coronary revascularization on the beating heart is an attractive alternative to conventional coronary artery bypass grafts (CCABG), but remains controversial. Our study compares the outcomes of consecutive patients undergoing off-pump CABG (OPCABG) with a group of similar patients undergoing consecutive CCABG. METHODS: A retrospective analysis of 268 patients who underwent elective CABG between July 1998 and July 1999 at St. Michael's Medical Center yielded 134 consecutive patients who underwent OPCABG and 134 consecutive patients who had CCABG. Patients' medical charts were reviewed for age, preoperative risk factors, operative findings, postoperative complications, and length of stay (LOS). RESULTS: The two cohorts were well matched, with similar ages (66.4 +/- 11.2 for OPCABG vs. 65.8 +/- 10 for CCABG, p = 0.66) and preoperative ejection fractions (EF) (44 +/- 13 vs. 44 +/- 12, p = 0.85). There were no hospital mortalities, and there were five conversions to cardiopulmonary bypass. The OPCABG group had a significantly shorter ICU and postoperative LOS. CONCLUSIONS: Our data suggests that a fair number of patients are potential candidates for OPCABG, the only contraindications being technical limitations or the surgeon's comfort level. Six- to twelve-month follow-up indicates that OPCABG can be performed safely with a decrease in LOS, and should be part of the surgeon's armamentarium.

Aged↗

Further evidence that 3-phosphoinositide-dependent protein kinase-1 (PDK1) is required for the stability and phosphorylation of protein kinase C (PKC) isoforms.

The multi-site phosphorylation of the protein kinase C (PKC) superfamily plays an important role in the regulation of these enzymes. One of the key phosphorylation sites required for the activation of all PKC isoforms lies in the T-loop of the kinase domain. Recent in vitro and transfection experiments indicate that phosphorylation of this residue can be mediated by the 3-phosphoinositide-dependent protein kinase-1 (PDK1). In this study, we demonstrate that in embryonic stem (ES) cells lacking PDK1 (PDK1-/- cells), the intracellular levels of endogenously expressed PKCalpha, PKCbetaI, PKCgamma, PKCdelta, PKCepsilon, and PKC-related kinase-1 (PRK1) are vastly reduced compared to control ES cells (PDK1+/+ cells). The levels of PKCzeta and PRK2 protein are only moderately reduced in the PDK1-/- ES cells. We demonstrate that in contrast to PKCzeta expressed PDK1+/+ ES cells, PKCzeta in ES cells lacking PDK1 is not phosphorylated at its T-loop residue. This provides the first genetic evidence that PKCzeta is a physiological substrate for PDK1. In contrast, PRK2 is still partially phosphorylated at its T-loop in PDK1-/- cells, indicating the existence of a PDK1-independent mechanism for the phosphorylation of PRK2 at this residue.

3-Phosphoinositide-Dependent Protein Kinases↗

The role of 3-phosphoinositide-dependent protein kinase 1 in activating AGC kinases defined in embryonic stem cells.

BACKGROUND: Protein kinase B (PKB), and the p70 and p90 ribosomal S6 kinases (p70 S6 kinase and p90 Rsk, respectively), are activated by phosphorylation of two residues, one in the 'T-loop' of the kinase domain and, the other, in the hydrophobic motif carboxy terminal to the kinase domain. The 3-phosphoinositide-dependent protein kinase 1 (PDK1) activates many AGC kinases in vitro by phosphorylating the T-loop residue, but whether PDK1 also phosphorylates the hydrophobic motif and whether all other AGC kinases are substrates for PDK1 is unknown. RESULTS: Mouse embryonic stem (ES) cells in which both copies of the PDK1 gene were disrupted were viable. In PDK1(-/-) ES cells, PKB, p70 S6 kinase and p90 Rsk were not activated by stimuli that induced strong activation in PDK1(+/+) cells. Other AGC kinases - namely, protein kinase A (PKA), the mitogen- and stress-activated protein kinase 1 (MSK1) and the AMP-activated protein kinase (AMPK) - had normal activity or were activated normally in PDK1(-/-) cells. The insulin-like growth factor 1 (IGF1) induced PKB phosphorylation at its hydrophobic motif, but not at its T-loop residue, in PDK1(-/-) cells. IGF1 did not induce phosphorylation of p70 S6 kinase at its hydrophobic motif in PDK1(-/-) cells. CONCLUSIONS: PDK1 mediates activation of PKB, p70 S6 kinase and p90 Rsk in vivo, but is not rate-limiting for activation of PKA, MSK1 and AMPK. Another kinase phosphorylates PKB at its hydrophobic motif in PDK1(-/-) cells. PDK1 phosphorylates the hydrophobic motif of p70 S6 kinase either directly or by activation of another kinase.

3-Phosphoinositide-Dependent Protein Kinases↗