Artefactual cleavage of E coli H-NS by OmpT

In the bacterium Escherichia coli, H-NS-(H1, H1a) is a heat-stable protein with a molecular mass of 15.5 kDa involved in nucleoid organisation and gene regulation linked to certain signal transduction pathways. We have shown that, following addition of preparations of everted inner membrane vesicles, heat-stable cleavage products of approximately 10 kDa of H-NS are formed in vitro from newly synthesised, radio-labelled H-NS and from purified H-NS. The 15.5 kDa protein and its cleavage products were also recovered from a minicell system. These results raised the possibility that cleavage of H-NS is physiologically significant. However, the cleavage of H-NS observed appears to occur during cell breakage and to depend on the method of protein extraction and the presence of the outer membrane protease, OmpT. Nevertheless, the results indicate that H-NS may contain at least two separate domains with cleavage occurring between these domains at a preferred OmpT site. Failure to take account of H-NS cleavage in sample preparation and analysis can lead to serious underestimation of H-NS levels.     

Coaggregation of oral lactobacilli with streptococci from the oral cavity

The ability of oral lactobacilli to coaggregate with streptococci and actinomycetes was investigated. Of the 7 species of lactobacilli studied, only two were capable of coaggregation and the coaggregation was restricted to streptococci. Lactobacillus salivarius strains (2/4) coaggregated with Streptococcus salivarius, Streptococcus gordonii, Streptococcus crista and tufted Streptococcus sanguis II strains. Lactobacillus fermentum (2/3) coaggregated with S. gordonii and S. sanguis. The coaggregation between L. salivarius and S. salivarius, S. gordonii or tufted S. sanguis II strains was mediated by a protein on the surface of the lactobacilli and was not inhibited by lactose. The coaggregation between L. fermentum and the streptococci was mediated by protein on the surface of the streptococci and was inhibited by lactose.     

Quantitative evaluation of pancreatic enhancement during dual-phase helical CT

PURPOSE: To determine the improvement in pancreatic enhancement at helical computed tomography (CT) performed with an early delay after administration of contrast material compared with that performed with a standard delay. MATERIALS AND METHODS: Dual-phase helical CT of the abdomen was performed in 120 patients with a 150-mL bolus of contrast material infused at 5 mL/sec. Early and standard delayed scanning was performed beginning at 20 seconds and 49-71 seconds, respectively. Regions of interest were measured in the head, body, and tail of the pancreas in 92 patients. The difference in enhancement between early and standard delayed scanning was calculated. RESULTS: Mean pancreatic enhancement was 82 HU +/- 3 (standard error) with an early delay, whereas enhancement on standard delay scans was 62 HU +/- 2 (P < .001). An improvement in enhancement greater than 10 HU was attained in 66 of 92 cases (72%). CONCLUSION: Pancreatic enhancement at helical CT with an early delay after contrast material administration is often significantly greater than the enhancement seen with a standard delay when a monophasic, rapidly infused bolus of contrast material is used.     

Improved accuracy of burn wound assessment using laser Doppler

The utility of the laser Doppler for determining burn depth has been questioned because of problems with technology and methodology. This study prospectively evaluates the ability of a new laser Doppler technique to predict burn healing time. Using the Periflux System 4000 laser Doppler, readings were taken on 305 burns (147 patients) on postburn day 3 or 4. Sixty-six wounds were used to derive a predictive function (phase I) and 152 wounds were used to test the function (phase II). Blood flow dynamics (flux), microvascular dilation capacity of the wounds to beat stress, and flow motion wave pattern (vasomotion) were studied using the laser Doppler, and seven parameters were evaluated to determine their relative contribution to the prediction of healing time. These parameters are hyperemic flux (flux value after heating to 42 degrees C), average hyperemic wave amplitude (AHWA), number of average flux units >100(F100), number readings with wave amplitude 75 (A5), average flux change (AFC), percentage of average flux increase, and relative flow capacity (RFC = AFC/average hyperemic flux). After readings were made, the wounds were observed and divided into two groups: those that healed in less than 14 days and those that healed or were grafted after 14 days. A step-wise discriminant analysis was used to assess the relative contribution of the Doppler-derived measures to healing time prediction. AHWA, F100, and RFC were included in the final discriminant function explaining 72% of the healing time variance (Wilks' lambda value 0.28; p value <0.0001). Predicted outcome = 0.05(AHWA) + 0.31(F100) + 5.0(RFC) - 2.3. With this derived function, there is 94% accuracy in the prediction of burn wound healing time compared with a physician predictive accuracy of 70%.     

Abnormal behavioral responses to fenfluramine in patients with affective and personality disorders. Correlation with increased serotonergic responsivity

Serotonergic responsivity was assessed in 20 psychiatric patients by the prolactin response to a fenfluramine challenge test. During the fenfluramine challenge 6 of 20 patients (30%) spontaneously reported psychopathologic reactions that included: increased anxiety/agitation, psychotic symptoms, illusions, mood elevation, and anergia. The time of peak behavioral symptoms (2.5 +/- 0.8 hrs) corresponded closely to the time of peak increase in prolactin levels (3.0 +/- 1.1 hr). Abnormal behavioral responders had statistically significant greater increases in prolactin 1 to 4 hr after fenfluramine when compared to normal responders. Patients who developed an abnormal psychopathologic response to fenfluramine were characterized by higher levels of anxiety and agitation at the time of admission to the hospital but otherwise were not distinguishable on the basis of severity of other psychiatric symptoms. This study suggests that increased serotonergic transmission may trigger anxiety, psychosis, and mood elevation in specific vulnerable individuals, whereas other patients with similar psychiatric illnesses are not affected.     

ELISA detection of IL-1 beta in human sera needs independent confirmation. False positives in hospitalized patients

Electrical field stimulation evoked a reproducible outflow of calcitonin gene-related peptide-like immunoreactivity (CGRP-LI) from the dorsal half of the rat spinal cord, an effect which was abolished by prior application of capsaicin, tetrodotoxin or removal of extracellular Ca2+. Adenosine (EC50 3.2 microM) and the selective adenosine A1 receptor agonist N6-cyclohexyladenosine (EC50 8.2 nM) inhibited evoked CGRP-LI outflow, while the selective adenosine A2 receptor agonist CGS-21680 was ineffective up to 10 microM. The action of adenosine was prevented by the adenosine A1 receptor selective antagonist 8-cyclopentyl-1,3-dipropylxanthine (30 microM), which did not affect CGRP-LI release on its own.     

Vesicoureteral reflux and ureteropelvic junction obstruction: association, treatment options and outcome

PURPOSE: We investigated the association, treatment options and outcomes of patients with ureteropelvic junction obstruction and concomitant vesicoureteral reflux. MATERIALS AND METHODS: We analyzed 6,790 consecutive pediatric urology records at our university. Treatment options included observation, and primary pyeloplasty, ureteroneocystostomy and nephroureterectomy. Hydronephrosis, reflux and obstruction were judged as resolved, improved, unchanged or worse. RESULTS: A total of 1,140 patients had vesicoureteral reflux, 224 had ureteropelvic junction obstruction and 41 had both conditions (39 ipsilateral and 6 contralateral kidneys). There was no increased risk of obstruction in patients with reflux when all grades of reflux were grouped (odds ratio 1.26, confidence interval 0.91 to 1.71). In contrast, subgroup analysis of patients with high grade reflux demonstrated a 5-fold increased risk of obstruction (odds ratio 5.0, confidence interval 2.4 to 10.8). One patient was lost to followup. Observation of 6 kidneys led to resolution of reflux in 3 (50%), resolution of obstruction in 3 (50%) and resolution or improvement of hydronephrosis in 4 (67%). Primary pyeloplasty was done on 29 kidneys with 10 (35%) requiring subsequent ureteroneocystostomy. At latest followup hydronephrosis resolved or improved in 24 patients (83%), vesicoureteral reflux resolved or improved in 19 (66%) and ureteropelvic junction obstruction resolved in all. Primary ureteroneocystostomy was performed on 5 kidneys, all of which required subsequent pyeloplasty. Hydronephrosis resolved in 3 patients (60%), and reflux and obstruction resolved in all. Two patients treated with primary nephroureterectomy, and 1 who underwent concomitant pyeloplasty and ureteroneocystostomy have had no subsequent urological problems. One patient awaits primary pyeloplasty. CONCLUSIONS: High grade vesicoureteral reflux is associated with ureteropelvic junction obstruction. No association with low or intermediate grade reflux was demonstrated. While some patients may be monitored expectantly, in our series pyeloplasty or nephrectomy was required in 81% and ureteroneocystostomy was required in 36%. In no case did primary ureteroneocystostomy protect against the subsequent need for pyeloplasty.     

Stability of proximal femoral grafts in canine hip arthroplasty

In a canine model, the fixation stability of a prosthesis and proximal bone graft composite were measured relative to the distal femur. One group had the prosthesis graft composite cemented into the distal femur. The second group had the prosthesis graft composite press fit into the distal femur for biologic ingrowth. Displacements of the proximal femoral grafts relative to the host bone in each group were measured after ex vivo (acute with graft) implantation and 4 months after implantation. A third group with no osteotomy (acute intact) simulated perfect graft to host bone union. Relative displacements representing 6 degrees freedom (translation and rotation) were calculated from the displacement values measured by 9 eddy current transducers. Measurements of displacement were used to test the hypothesis that distal press fit fixation equals distal cement fixation at 4 months after implantation. In all cases the measured translations and rotations of the graft to implant construct were small and of a magnitude that should encourage bone ingrowth (< 0.05 mm and < 0.1 degree, respectively). The stability of the press fit group at 4 months was not significantly different from the cemented group in axial and transverse displacement during axial and transverse loading, respectively. There was no difference in stabilities at 4 months between distal press fit and cemented fixation in hip replacements requiring a proximal femoral graft.     

Prostate enlargement in mice due to fetal exposure to low doses of estradiol or diethylstilbestrol and opposite effects at high doses

On the basis of results of studies using high doses of estrogens, exposure to estrogen during fetal life is known to inhibit prostate development. However, it is recognized in endocrinology that low concentrations of a hormone can stimulate a tissue, while high concentrations can have the opposite effect. We report here that a 50% increase in free-serum estradiol in male mouse fetuses (released by a maternal Silastic estradiol implant) induced a 40% increase in the number of developing prostatic glands during fetal life; subsequently, in adulthood, the number of prostatic androgen receptors per cell was permanently increased by 2-fold, and the prostate was enlarged by 30% (due to hyperplasia) relative to untreated males. However, as the free serum estradiol concentration in male fetuses was increased from 2- to 8-fold, adult prostate weight decreased relative to males exposed to the 50% increase in estradiol. As a model for fetal exposure to man-made estrogens, pregnant mice were fed diethylstilbestrol (DES) from gestation days 11 to 17. Relative to controls, DES doses of 0.02, 0.2, and 2.0 ng per g of body weight per day increased adult prostate weight, whereas a 200-ng-per-g dose decreased adult prostate weight in male offspring. Our findings suggest that a small increase in estrogen may modulate the action of androgen in regulating prostate differentiation, resulting in a permanent increase in prostatic androgen receptors and prostate size. For both estradiol and DES, prostate weight first increased then decreased with dose, resulting in an inverted-U dose-response relationship.