Fibrinogen degradation products in patients with the Quebec platelet disorder

The Quebec platelet disorder is a serious bleeding disorder associated with proteolysis of α-granular proteins, including fibrinogen. We evaluated fibrinogen degradation product (FDP) assays as screening tests for this disorder. Patients with the Quebec platelet disorder (13/13) had elevated serum FDPs ( P  <0.01) due to secreted degraded platelet fibrinogen, but normal plasma FDPs and D-dimers. Unrelated controls with bleeding disorders (32/34) had undetectable FDPs, and controls with FDPs due to disseminated intravascular coagulation (DIC) and other illnesses (11/11) had elevated FDPs in all assays ( P  < 0.01). Immunoblot analyses indicated plasma fibrinogen was normal in the Quebec patients and platelet FDPs were found in their platelet lysates, releasates and serum samples. Their platelet FDPs were not altered by treatment with fibrinolytic inhibitors and were different from the FDPs in DIC and in plasmin-digested fibrinogen. Tests for serum FDPs may provide a simple rapid way to screen for the Quebec platelet disorder.     

Cerebral blood flow asymmetry in the detection of extracranial cerebrovascular disease

Regional cerebral blood flow was measured by the 133Xe inhalation technique in patients with unilateral carotid occlusion, unilateral carotid occlusion and contralateral carotid stenosis, bilateral carotid occlusion, or normal arteriograms. After adjusting for age, sex, and history of stroke, hemispheric blood flow asymmetry was shown to be a predictor of unilateral carotid occlusion with a sensitivity of 80.6% and a specificity of 80.5%. Asymmetry of regional cerebral blood flow is useful in the assessment of patients with extracranial cerebrovascular disease.     

Use of the Membrane Invasion Culture System (MICS) as a screen for anti-invasive agents

The Membrane Invasion Culture System (MICS) assay was adapted for relatively rapid screening of compounds and used to identify anti-invasive drugs that inhibit human and murine tumor cell migration through a reconstituted basement membrane in vitro. Cell lines demonstrating low and high invasive and metastatic potentials were tested with all compounds for tumoricidal effects prior to evaluation in MICS at non-cytotoxic doses. The effect on invasive potential in the MICS assay was determined in 3 categories: (1) 48 hr drug pre-treatment prior to seeding in the MICS (exceptions: 90 min pre-treatment with pertussis toxin and, for some studies, continuous exposure for 2-7 days); (2) peptide or prostaglandins 2 hr after seeding and attachment to the membranes in MICS followed by continuous exposure; and (3) cells receiving neither drug nor peptide treatment and serving as controls in each MICS chamber. Since invasion involves cellular motility and deformability, some cytoskeleton disrupting agents were selected. Of these, vincristine, colcemid and colchicine inhibited invasion but taxol did not. Pre-treatment with cAMP agonists produced conflicting results: dibutyryl cAMP and 8-(4-chloro-phenylthio) cAMP resulted in 50% and 38% reduction in invasion, respectively, whereas 8-bromo cAMP stimulated invasive potential by 30%. Forskolin and cholera toxin both significantly reduced invasiveness. Pre-treatment with 5-azacytidine and araC, to consider the role of methylation and proliferations decreased invasive ability. Anti-metastatic drugs such as gamma-interferon and razoxane inhibited invasive potential but to varying degrees. Treatment of cells with prostaglandins E2, F2 alpha, A2, and D2 were ineffectual; however, indomethacin mildly inhibits invasion (less than 30%).(ABSTRACT TRUNCATED AT 250 WORDS)     

Applications of cellular fatty acid analysis.

More than ever, new technology is having an impact on the tools of clinical microbiologists. The analysis of cellular fatty acids by gas-liquid chromatography (GLC) has become markedly more practical with the advent of the fused-silica capillary column, computer-controlled chromatography and data analysis, simplified sample preparation, and a commercially available GLC system dedicated to microbiological applications. Experience with applications in diagnostic microbiology ranges from substantial success in work with mycobacteria, legionellae, and nonfermentative gram-negative bacilli to minimal involvement with fungi and other nonbacterial agents. GLC is a good alternative to other means for the identification of mycobacteria or legionellae because it is rapid, specific, and independent of other specialized testing, e.g., DNA hybridization. Nonfermenters show features in their cellular fatty acid content that are useful in identifying species and, in some cases, subspecies. Less frequently encountered nonfermenters, including those belonging to unclassified groups, can ideally be characterized by GLC. Information is just beginning to materialize on the usefulness of cellular fatty acids for the identification of gram-positive bacteria and anaerobes, despite the traditional role of GLC in detecting metabolic products as an aid to identification of anaerobes. When species identification of coagulase-negative staphylococci is called for, GLC may offer an alternative to biochemical testing. Methods for direct analysis of clinical material have been developed, but in practical and economic terms they are not yet ready for use in the clinical laboratory. Direct analysis holds promise for detecting markers of infection due to an uncultivable agent or in clinical specimens that presently require cultures and prolonged incubation to yield an etiologic agent.     

STIs in women: symptoms and examination

STIs are common, particularly in young women. Sexual history-taking is eased by private and comfortable surroundings, establishing a good professional relationship, having a non-judgemental attitude, and choosing words that are both appropriate to the consultation and readily understood by the woman. Genital examination is facilitated by privacy, careful and sensitive technique and the availability of the right equipment, including a gynaecological examination couch and a range of speculums. The screening investigations selected depend on the setting, local availability of investigations and prevalence of infections, and the presentation and wishes of the client. Sampling requirements are changing with the advent of new diagnostic techniques such as nucleic acid amplification tests for Chlamydia; these can be undertaken on samples taken by the woman herself (e.g. perineal swabs, urine), thereby avoiding the need for speculum examination in some circumstances.     

Platelet glycine, glutamate and aspartate in primary headache

Platelet levels of glutamic and aspartic acid and glycine were measured in patients with migraine with aura, migraine without aura, tension headache and cluster headache. High levels of these amino acids were found in patients with migraine with aura compared to normal subjects and other headache groups. During headache, glutamate levels further increased in migraine with aura patients. These findings may have relevance to the neurological symptoms of migraine with aura.     

Interactions between serotonin, thyrotropin-releasing hormone, and substance P in the CNS regulation of adrenocortical secretion

Double-labeling immunohistochemical studies were performed to discern the morphological relationships between corticotropin-releasing factor-immunoreactive (CRF-ir) perikarya and afferent innervation in the hypothalamic paraventricular nucleus (PVN) of the rat. Attention was focussed on the local innervation by serotonin (5-hydroxytryptamine, 5-HT), thyrotropin-releasing hormone (TRH) and substance P (SP)-ir nerve terminal fibers. 5-HT-ir and SP-ir fibers were present in moderate numbers, in close apposition with CRF-ir perikarya. Sparse TRH-ir fibers were observed, but a population of TRH-ir perikarya was found in proximity with the CRF-ir cell bodies. TRH-ir perikarya in the PVN were surrounded by both 5-HT- and SP-ir fibers. Neuroendocrine studies were performed to investigate the interactions between 5-HT, TRH and SP in the regulation of hypothalamo-pituitary-adrenocortical (HPA) secretion. Male rats were prepared bearing cannulae for intracerebroventricular (ICV) or intra-PVN administration of drugs. 5-HT, at all doses tested (0.1, 40, or 80 nmol, ICV), caused increases in plasma corticosterone (CS) concentrations in tail-vein blood collected 20 min after injection. ICV injections of TRH caused dose-dependent increases in plasma CS, but did not further increase HPA responses when injected together with 5-HT. SP alone had little effect, although a significant reduction in plasma CS concentrations was observed in several individual experiments. However, SP (0.1 nmol) significantly attenuated CS responses following high doses of 5-HT (40 and 80 nmol, ICV), although the response to 0.1 nmol 5-HT was not affected. Combined injection of SP with TRH resulted in HPA responses not different from those following TRH alone. Similarly, SP did not reduce the HPA response observed with TRH and 40 nmol 5-HT in combination. Intra-PVN injections of 5-HT (0.1 or 40 nmol) and TRH also increased plasma CS concentrations. Intra-PVN injections of SP had little effect on plasma CS concentrations although a tendency toward a decrease in plasma CS was observed, as with the ICV injections. Combined intra-PVN injection of 5-HT (0.1 nmol) with TRH (0.1 nmol) did not significantly alter the response compared with that observed following TRH alone, although plasma CS concentrations were greater than with 0.1 nmol 5-HT. Combined intra-PVN injections of SP (0.1 nmol) with 5-HT (0.1 nmol) resulted in a significant decrease in plasma CS concentration compared with that following 5-HT alone, but SP did not prevent the CS response to a higher dose of 5-HT (40 nmol). Similar to the results observed following ICV administration, SP in combination with TRH did not affect the HPA response observed with TRH, or with 5-HT and TRH in combination. The results of these studies demonstrate that 5-HT and TRH serve to stimulate adrenocortical secretion, both at the level of the PVN and at extrahypothalamic structures, but that SP exerts an inhibitory influence that is capable of overcoming some of the stimulatory effects of 5-HT.     

Platelet deposition on vascular grafts. The accuracy of in vivo quantitation and the significance of in vivo platelet reactivity.

An in vivo platelet imaging system utilizing indium-111-labeled platelets and technetium-99m-labeled red cells was used to serially study and compare platelet deposition on autologous external jugular vein grafts, autologous arterial grafts, polytetrafluoroethylene (Gore-tex) and two Dacron (Meadox and USCI) small diameter (4mm) vascular grafts implanted end-to-end in canine carotid and femoral arteries. This method of quantitating platelet deposition was validated by correlating deposition measured in vivo with deposition measured directly on explanted grafts (r = 0.94, p less than 0.01). Platelet accumulation on all grafts was greatest immediately after implantation and declined over time. None of the artery or vein grafts thrombosed, and they had the lowest level of platelet deposition at all times. Platelet deposition on Gore-tex grafts was significantly less than on USCI Dacron grafts from 24 hours to 1 month after implantation. There was no statistical difference in 1-month patency among the synthetic graft groups. Synthetic grafts that thrombosed during the first month accumulated significantly more platelets immediately after operation than did those grafts that remained patent. Patent Dacron grafts with low levels of platelet deposition had less thrombotic debris at explantation on the luminal surface than did those grafts with high levels of platelet deposition. Differences in initial platelet deposition appeared to be more a function of platelet reactivity within each dog rather than the material used in graft construction.