Differential effects of specific opioid receptor agonists on rat pup isolation calls.

The possibility was investigated that specific opioid receptor types might selectively alter the production of isolation-induced ultrasonic vocalizations. Intracisternal injections of mu, delta and kappa opioid receptor agonists were administered to isolated 10-day-old rat pups. The mu receptor agonist [D-Ala2-NMe-Phe4-Gly-ol]-enkephalin (DAMGO) and delta receptor agonist [D-Pen2, D-Pen5]-enkephalin (DPDPE) both reduced the rate of isolation-induced ultrasonic calling in the absence of sedation. The kappa receptor agonist U50,488 had the opposite effect, significantly raising the rate of vocalization. Fourteen-day-old pups, with a larger delta receptor population, showed a greater sensitivity to DPDPE than was seen in the younger animals.     

Adrenergic Receptor Genotype but Not Perioperative Bisoprolol Therapy May Determine Cardiovascular Outcome in At-risk Patients Undergoing Surgery with Spinal Block: The Swiss Beta Blocker in Spinal Anesthesia (BBSA) Study: A Double-blinded, Placebo-controlled, Multicenter Trial with 1-Year Follow-up

Background: Neuraxial blockade is used as primary anesthetic technique in one third of surgical procedures. The authors tested whether bisoprolol would protect patients at risk for cardiovascular complications undergoing surgery with spinal block.

Methods: The authors performed a double-blinded, placebo-controlled, multicenter trial to compare the effect of bisoprolol with that of placebo on 1-yr composite outcome including cardiovascular mortality, nonfatal myocardial infarction, unstable angina, congestive heart failure, and cerebrovascular insult. Bisoprolol was given orally before and after surgery for a maximum of 10 days. Adrenergic receptor polymorphisms and safety outcome measures of bisoprolol therapy were also determined.

Results: A total of 224 patients were enrolled. Spinal block could not be established in 5 patients. One hundred ten patients were assigned to the bisoprolol group, and 109 patients were assigned to the placebo group. The mean duration of treatment was 4.9 days in the bisoprolol group and 5.1 days in the placebo group. Bisoprolol therapy reduced mean heart rate by 10 beats/min. The primary outcome was identical between treatment groups and occurred in 25 patients (22.7%) in the bisoprolol group and 24 patients (22.0%) in the placebo group during the 1-yr follow-up (hazard ratio, 0.97; 95% confidence interval, 0.55-1.69; P = 0.90). However, carriers of at least one Gly allele of the [beta]1-adrenergic receptor polymorphism Arg389Gly showed a higher number of adverse events than Arg homozygous (32.4% vs. 18.7%; hazard ratio, 1.87; 95% confidence interval, 1.04-3.35; P = 0.04).     

Successful regrafting of a transplanted liver

We report on the successful regrafting of a transplanted liver. The donor liver was first grafted into a patient suffering from cryptogenic cirrhosis; the patient died 1 day after the elective transplantation of cerebral bleeding. The well-functioning graft was harvested again and transferred to our institution. After another 12 h of cold ischemia, the liver was reperfused in an urgently registered patient with recurrence of hepatitis B in his first graft. The transplantation was successfully performed and the patient is now doing well, more than 5 months after regrafting with the reused liver. Received: 21 October 1996 Received after revision: 9 January 1997 Accepted: 27 January 1997     

Unstable fractures of the upper thoracic spine

Unstable fractures of the upper thoracic spine are a therapeutic challenge because of the high rate of associated injuries and frequent occurrence of spinal cord lesions. This study focuses on the operative treatment of these injuries. We present nine patients with unstable injuries of the upper thoracic spine. According to the classification of Magerl et al., there are seven type C and two type B fractures. Eight patients had complete transverse lesions of the cord and one had no evidence of spinal cord injury. All patients presented associated injuries: nine patients had fractured ribs, three a fractured sternum and three a severe haemothorax. All underwent operative treatment: six posterior fusions only, two anterior interbody fusions only after thoractotomy owing to severe bleeding from ruptured intercostal arteries, and one a combined fusion after failure of posterior fusion. The treatment of these patients should be individualized, depending on the associated injuries, loss of blood owing to fractured spine and ruptured vessels and depending on the back pain resulting from an unstable thorax. In general, we prefer the posterior approach to unstable fractures of the upper thoracic spine.     

Alterations of the immune system following splenectomy in childhood

After splenectomy due to blunt abdominal trauma, splenectomized children showed a restricted pattern of T-cell immunodeficiency compared to age and sex-matched normal children. Peripheral blood total (CD3) T-cell counts of 11 splenectomized children of 43%, double positive helper (CD4) inducer subpopulation (CD29) cell counts of nine splenectomized children of 7%, and phytohemagglutinin (PHA)-induced T-cell proliferation of 11 splenectomized children of 53,206 c.p.m. were significantly (p less than 0.05) lower than values of normal children (61% CD3 cells, n = 12; 13% CD4CD29 cells, n = 11; 107,832 c.p.m. PHA-induced proliferation, n = 12). The deficit of CD4CD29 cell numbers may be due to impaired maturation of these particular CD4 lymphocytes and may explain diminished PHA-induced proliferation in small children. The significantly higher B-lymphocyte counts of splenectomized children (21%, n = 11; 558 cells/mm3, n = 10) compared with 12 normal children (14%; 329 cells/mm3) may be due to loss of the reservoir function of the spleen.     

A new technique for reconstruction of the common bile duct after severe injury by laparoscopic cholecystectomy

Zusammenfassung Schwere Gallengangsverletzungen sind nach laparoskopischen häufiger als nach konventionellen Cholezystektomien zu beklagen. Sie resultieren meist aus technischen Problemen oder unzureichender Identifikation der entsprechenden Strukturen. Das sofortige Erkennen der Gallengangsverletzung ist fur eine entsprechende Therapie ebenso wichtig wie die ädequate chirurgische Technik, die angewendet wird. Hier wird eine neue Methode der Gallenwegrekonstruktion unter Verwendung eines Jejunalkonduits vorgestellt. Das Verfahren wurde bislang bei 5 Patienten mit schweren Gallengangsverletzungen (Typ 3 and 4 nach Bismuth) [2] nach laparoskopischer Cholezystektomie erfolgreich angewendet. Der Verlauf nach nunmehr 9 Monaten bis zu mehr als 2 Jahren war ohne Probleme and vielversprechend. Die Methode der Anwendung des Jejunalkonduits erscheint uns zur Rekonstruktion bei schweren Gallengangverletzungen sehr empfehlenswert.

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A new technique for reconstruction of the common bile duct after severe injury by laparoscopic cholecystectomy

Laparoscopic cholecystectomy involves a higher incidence of severe common bile duct injury than did open cholecystectomy. The severe injuries most often result from technical problems and inadequate exposure. Reconstruction of the bile duct is then possible provided that an immediate diagnosis is made and an appropriate surgical technique is applied. The report focuses on a new method of reconstruction of the common bile duct by interposition of a small jejunal conduit. The procedure was performed in five patients with severe bile duct injury (Bismuth class 3–4) that occurred during laparoscopic cholecystectomy. The outcome after follow-up periods of 9 months to more than 2 years is promising. This method of reconstruction is therefore recommended for severe forms of bile duct injury.

     

Development of an ELISA for detection of an organophosphorus compound using monoclonal antibodies

This paper describes a specific and highly sensitive ELISA system using monoclonal antibodies in order to assay an organophosphorus compound. The soman derivative methyl phosphonic acid, p-aminophenyl 1,2,2,-trimethyl-propyl diester (MATP) served as model substance. In order to obtain antibody-producing hybridomas BALB/c mice were immunized with MATP linked onto human serum albumin (HSA). The spleen cells of immunized mice were fused with syngenic plasmacytomas of the non-producer-line X63Ag8.653 with the aid of polyethylene glycol. To eliminate undesirable cross-reaction, common screening procedures were modified by directly coating the ELISA plates with hapten. Five out of 15 positive cell-lines were cloned by limiting dilution and further propagated. The respective immunoglobulin class and subclass of the obtained monoclonal antibodies was determined. Four of which were identified as IgG₁, the other as IgG_2a. After enrichment of antibodies in ascites and their isolation by protein A-sepharose, the affinity of various monoclonal antibodies was estimated in competitive inhibition enzyme immunoassay (CIEIA) by measuring the IC₅₀ rates of free MATP. The rates were found to lie between 2.5 × 10^–6 mol/l and 4.3 × 10^–4 mol/l MATP. The IC₁₀ rate for detectable MATP concentration was 5.4 × 10^–7 mol/l MATP. Test duration was 280 min. The reactivity of the monoclonal antibodies with structurally related substances was used to check their specificity. Cross-reaction turned out to be negative. In order to develop a direct competitive ELISA, MATP was linked to horse radish peroxidase (HRPO) by adding a spacer. This helped to reduce total duration to 40 min. The detection level was further reduced to 1.3 × 10^–7 mol/l MATP (corresponding to 975 pg/25 l test-buffer) using the monoclonal antibody F71D7. Likewise, MATP was detected in goat serum, chicken serum, rabbit serum, milk and company's water in concentrations between 2.1 × 10^–7 mol/l (IC₁₀, company's water) and 4.9 × 10^–8 mol/l (IC₁₀, milk).     

Maternal effects in infant and adult phenotypes of 5HT1A and 5HT1B receptor knockout mice

The influence of the pre- and postweaning maternal environment on the offspring's phenotype was examined in 5-HT1A and 5-HT1B receptor knockout mice (KO1A and KO1B, respectively). We have previously shown that, when born to and raised by homozygous dams of the same genotype, adult KO1A are more anxious than wild-type (WT) mice, and adult KO1B are hyperactive and slightly less anxious than WT mice. We extend our studies here to the behavioral results of the offspring's own genotype, when the dam's genotype is constant, and the effects of the dam's genotype when the offspring's genotype is constant. In Experiments 1 and 2, KO1A-/- pups produced less ultrasonic vocalizations (USV) than controls in an isolation test on postnatal Day 7 when born to and reared by KO1A dams, either -/- or +/-. Heterozygous F1 pups reared by KO1A-/- dams produced more USV and were less anxious in the plus-maze at 2 to 3 months of age than F1 pups born to and reared by WT dams (Experiment 3). F1 pups reared by KO1B-/- dams produced less USV and were more anxious in the plus-maze than F1 pups reared by WT dams (Experiment 4). The results support a role for maternal effects that may comprise direct effects such as the dam's behavior and nutritional care of the pup, and possibly more complex indirect effects through the establishment of idiosyncratic dam-pup dyadic interactions. We recommend that breeding techniques that rely on same genotype (mutant-mutant or WT-WT) breeding pairs not be used to generate offspring when the focus of research is the study of gene function, but rather when familial effects need to be studied.     

Electroretinography in rats

Basic mechanisms of the rat ERG were reinvestigated with contemporary methods of recording and photic stimulation via an optic fiber system connected with a contact lens. Flash stimulation and background illuminance were performed with photometrically defined light stimuli. ERGs recorded in darkness, from dark adapted rats, were similar with those observed in earlier work in which light flashes of much longer duration had been used. Flash stimulation carried out under stepwise increased background illuminance gave new information on the characteristics of oscillatory potentials of the ERG. In general the present observations agreed with the notion that the rods of the rat retina are as sensitive as in man, whereas the cones are functionally less efficient with respect to light sensitivity and temporal resolution. Differences in function of the cones from rat to man have to be kept in mind when using the rat ERG as a model for risk assessment in safety studies.