A capture enzyme-linked immunosorbent assay for species-specific detection of Bothrops venoms

A direct sandwich enzyme-linked immunosorbent assay (ELISA), employing affinity purified antivenom antibodies specifically recognizing the homologous venom, was developed for species-specific detection of bothropic venom. The method is based on a two-step affinity purification of the specific antibodies. A species monovalent antivenom is adsorbed onto a venom adsorbent containing heterologous venoms from the Bothrops, Crotalus and Lachesis genera. The species-specific antibodies obtained, are then adsorbed onto a second venom adsorbent containing only the homologous venom for the removal of non antivenom antibodies. Venom concentrations of 0.1 and 1,000 ng/ml were specifically identified for Bothrops jararacussu and B. alternatus venom respectively.     

Diagnosis and surgery of brachial plexus injuries

The surgical outcome of traumatic injuries of the brachial plexus (BP) depends on the following parameters: 1) accurate preoperative diagnosis of cervical root avulsion; 2) time interval between injury and surgery; 3) delicate handling of the nerve tissue; and 4) postoperative physiologic training. This report is based on a 15-year experience in brachial plexus surgery and is supported on the grounds of two major studies. In a prospective study, the authors controlled for the reliability of preoperative radiologic diagnosis by myelo-CT and MRI scans for 40 patients, to evaluate the integrity of the intraspinal cervical roots after brachial plexus injury. Surgical inspection via a cervical hemilaminectomy proved the accuracy of 85 percent and 52 percent of CT myelography and MRI, respectively. Retrospective statistical analyses were carried out of the long-term surgical results of 54 patients with traumatic injuries of the BP who received a grafting procedure between cervical roots C5 or C6 and the musculocutaneous nerve. Patients operated on up to 6 months after trauma showed a better result than patients operated on later than 12 months after trauma (p<0.05). In contrast, grafting between cervical root C5 or C6 and the use of different sural-graft sizes to reconstruct the musculocutaneous nerve demonstrated no statistically significant difference in the final outcome.     

Platelet function abnormalities in a family with recurrent arterial thrombosis

Three young family members with recurrent arterial thrombosis underwent investigation for lipid or coagulation abnormalities. Lipoprotein electrophoresis, cholesterol, triglyceride levels, and routine coagulation studies were unremarkable. By contrast, testing of platelet function showed enhanced platelet aggregability to epinephrine and collagen in two of the subjects. In addition, release of 14C-serotonin by adenosine diphosphate and epinephrine was increased over control values in these same two patients. The third subject demonstrated decreased platelet aggregation and lowered 14C-serotonin release, but was symptomatic with rest pain at the time of testing. The ongoing in vivo thrombosis in the third subject may account for hypocoagulable platelets by in vitro testing. These abnormally sensitive platelets identified by platelet function testing may be associated with a familial "hypercoagulability" syndrome. Definition of the hemostatic abnormality in these individuals provided a rational basis for pharmacological therapy with antiplatelet drugs, which appeared to be successful.     

Quinoxaline derivatives: structure-activity relationships and physiological implications of inhibition of N-methyl-D-aspartate and non-N-methyl-D-aspartate receptor-mediated currents and synaptic potentials.

The inhibitory potencies at excitatory amino acid (EAA) receptors of 11 quinoxaline derivatives were evaluated in two-electrode voltage-clamp recordings of Xenopus oocytes injected with rat cortex mRNA. Currents activated by kainate or (RS)-alpha-amino-3-hydroxy-5-methyl-4-isoxazole-propionic acid (AMPA) in Xenopus oocytes were inhibited competitively by all the quinoxaline derivatives, with apparent Ki values ranging from 0.27 to 300 microM against kainate and from 0.25 to 137 microM against AMPA. An excellent correlation was observed between inhibitory potencies of the quinoxaline derivatives against kainate and AMPA currents, in support of the contention that in this preparation these two agonists act at a single site. All 11 quinoxaline derivatives also inhibited current activated by the combination of glycine and N-methyl-D-aspartate (NMDA), apparently acting at the glycine site, and did so over a narrower range of apparent Ki values (0.37-8.1 microM). The correlation between the quinoxalines' kainate/AMPA potencies and their glycine/NMDA potencies was relatively weak. Thus, the quinoxaline derivatives were all good antagonists of glycine/NMDA currents and displayed a greater range of potencies against kainate and AMPA. The inhibitory effects of the six quinoxaline derivatives most potent in the Xenopus oocyte experiments were also tested against the excitatory postsynaptic field potential (EPSFP) recorded in the pyramidal cell dendritic field of the CA1 region of hippocampal slices after stimulation of the Schaffer collateral-commissural pathways. In slices superfused with "normal" medium (containing 1 mM Mg2+), in which the EPSFP is mediated primarily by non-NMDA receptors, IC50 values correlated closely with the Ki values against kainate/AMPA obtained in oocyte experiments but were approximately 8-fold higher. Similarly, in slices superfused with nominally Mg(2+)-free medium, in which the EPSFP is amplified due to a relief of the Mg2+ block of NMDA receptors, IC50 values correlated closely with the Ki values against glycine/NMDA obtained in oocyte experiments but were 60-fold higher. This comparison of results from the two experimental systems lends further support to the argument that hippocampal synaptic transmission is mediated postsynaptically by kainate/AMPA-type and NMDA/glycine-type EAA receptors that are pharmacologically indistinguishable from those expressed in mRNA-injected Xenopus oocytes. Furthermore, it suggests that EAA receptors in situ may be nearly saturated by high local concentrations of the endogenous ligands, a condition that would contribute substantially to the apparent non-NMDA receptor selectivity of certain quinoxaline derivatives.     

The use of tapioca as coverage in viral titration]

Virus titration is an important step required on viral vaccines quality control. "Plaque assay", which employs several types of overlay media, is usually used on viral titrations. In this paper we describe the use of Tapioca as an overlay media. Firstly, the toxicity of Tapioca was tested on Vero cells inoculated or not with the Yellow Fever virus (YF) 17DD vaccine strain. Secondly, different batches of the 17DD virus using the Tapioca and Karaya gum as the overlay on Vero cells were tested when higher titres were obtained using Tapioca. Tapioca was also shown to be a suitable overlay to be used in thermostability and plaque reduction neutralization tests. Other systems could benefit from the use of Tapioca as an overlay, since it was possible to titer Measles virus in Vero cells. Tapioca is a cheap Brazilian product, is locally available, easy to use, and reliable. Its use is suggested.     

Chronic immobilization-induced stress increases plasma testosterone and delays testicular maturation in pubertal rats

We investigated whether chronic stress, applied from prepuberty to early puberty, interferes with the spermatogenic and androgenic testicular functions. Male pubertal rats (40 days old) were immobilized 6 h per day for 15 days. Plasma concentrations of corticosterone, prolactin and testosterone were significantly augmented following immobilization, whereas plasma luteinizing hormone decreased and follicle-stimulating hormone was not altered. Acute immobilization (5 min) increased prolactin and testosterone levels in control rats but caused a significantly higher increase in these hormones when superimposed on chronic stress. A lower extent of testicular maturation was observed in pubertal rats immobilized from prepuberty.     

Cystic schwannomas of the jugular foramen: clinical and surgical remarks

OBJECTIVE: The goals of this report were to outline the clinical presentation, radiological characteristics, surgical techniques, postoperative morbidity, and long-term follow-up results for cystic jugular foramen (JF) schwannomas and to describe their differences, compared with solid schwannomas involving the JF. METHODS: A retrospective analysis of radiological studies and surgical records identified five primarily cystic tumors among 21 cases of JF schwannomas that had been surgically treated at our institution. RESULTS: Two types of cystic JF schwannomas were observed, i.e., Type 1 lesions, which are single large cysts with thin ring-like enhancement of the tumor wall, and Type 2 lesions, which are multiple cysts with very irregular, thick enhancement of the cyst wall. The most common symptoms were hearing loss, ataxia, and headaches. Total surgical removal could be performed in all cases. The immediate postoperative findings indicated hearing improvement in three cases. No deterioration of lower cranial nerve function was observed. All patients were independent in the immediate postoperative period and in the long-term follow-up period (Karnofsky Performance Scale score, 90). CONCLUSION: Surgical treatment of cystic JF schwannomas can be very demanding because of generally stronger adhesion of the tumor capsule to the surrounding structures, fragile tumor capsules, and difficulty in identification of the arachnoidal planes in some cases. Early identification of the arachnoidal planes without opening of the cyst and sharp dissection may be useful. Careful intradural opening of the JF should be performed to achieve total removal of the last tumor portion within the JF. A comparison of these lesions with solid schwannomas involving the JF indicated that cystic tumors affected a younger population, with less preoperative swallowing impairment (P < 0.05). The immediate postoperative course in both types of cystic JF schwannomas was usually better than for solid lesions, because of minor postoperative cranial nerve morbidity, especially involving lower cranial nerve function, in the latter cases. Long-term follow-up data failed to demonstrate any significant differences in final patient outcomes, however.