Use of a high-throughput umu-microplate test system for rapid detection of genotoxicity produced by mutagenic carcinogens and airborne particulate matter

In the present study, we developed a rapid umu-microplate test system that uses the nitroreductase- and O-acetyltransferase-overproducing Salmonella typhimurium strain NM3009 and the O-acetyltransferase-overproducing S. typhimurium strain NM2009 to detect genotoxic activity in small volume samples. The assay was used to test the genotoxicity of several standard mutagens and environmental samples. Exponentially growing cultures of NM3009, NM2009, and the parental strain TA1535/pSK1002 were incubated in 96-well microplates with test chemicals both in the presence and in the absence of rat liver S9. The relative beta-galactosidase activities were then determined colorimetrically using either chlorophenol red-beta-D-galactopyranoside (CPRG) or O-nitrophenyl-beta-D-galactopyranoside (ONPG) as a measure of umuC gene induction activity. The sensitivities of NM3009 without S9 mix and NM2009 with S9 mix to nitroarenes and aromatic amines were up to 24- to 75-fold higher than those of the parent strain. Induction of umuC gene expression was detected more readily with CPRG than ONPG. The umu-microplate assay also detected genotoxicity in organic extracts of particulate matter from air samples collected in Osaka City, Japan. The pattern of the responses suggested that the genotoxic activity of the particulate extract was due primarily to nitrated polycyclic aromatic hydrocarbons. Our results indicate that the umu-microplate assay may be a useful way of carrying out rapid screens for genotoxicity in small-volume environmental samples.     

Dynamics of some indices of blood clotting after intraperitoneal injection of thrombin into noninbred albino rats

The possibility of giving thrombin by intraperitoneal injection as a test of the function of the blood clotting system (the in vivo thrombin test) was demonstrated in experiments on noninbred albino rats.Department of Biochemistry, Tyumen' Medical Institute. (Presented by Academician of the Academy of Medical Sciences of the USSR N. A. Fedorov.) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 86, No. 12, pp. 675–677, December, 1978.     

Sensitivity to mechanical stimuli and the role of general sensory and perceptual processes in heartbeat detection

Sensitivity to heartbeat sensations is commonly assessed using tasks that require individuals to judge the simultaneity of heartbeats and tones. In two experiments, we investigated the suitability of this paradigm for examining cardioception. In the first experiment, participants judged the simultaneity of near–threshold vibrations and suprathreshold tones. Precision in judging vibration–tone simultaneity was directly related to the detectability of the mechanical stimuli, thereby supporting use of the simultaneity paradigm to assess heartbeat detection. In the second experiment, we examined the influences of sensitivity to mechanical stimuli and the ability to make intermodality simultaneity judgments on the precision of heartbeat detection. We measured participants' vibrotactile thresholds, precision in judging light–tone simultaneity, and precision in judging heartbeat–tone simultaneity. The ability to judge the simultaneity of lights and tones accounted for 24.3% of the variance in precision of heartbeat detection, and mechanical sensitivity accounted for a further 8.5%.     

心脑血管病患者微血栓与D－2聚体、血小板聚集的相互关系

观察２０例急性脑梗塞，３０例冠心病在微循环中微血栓的变化，同时检测血小板聚集率、Ｄ－２聚体。结果表明微血栓阳性组血小板聚集率、Ｄ－２聚体均高于微血栓阴性组。治疗后二组血小板聚集、Ｄ－２聚体明显减少。提示微血栓、Ｄ－２聚体及血小板聚集率可作为评定冠心病、脑梗塞病情变化的一种指标     

In vitro mutagenicity of valepotriates

Valepotriates are epoxide-bearing triesters of the monoterpene alcohol 4,7-dimethylcyclopenta-(c)-pyrane isolated from the roots of several Valerianacae species. They are regarded as the main tranquilizing constituents of these drugs.Although the valepotriates valtrate/isovaltrate (VAL) and dihydrovaltrate (DH-VAL) showed a strong alkylating activity against the nucleophilic agent 4-(p-nitrobenzyl)-pyridine (NBP), they were not clearly mutagenic for the strains TA98, TA100, TA1535, and TA1537 of Salmonella typhimurium or for the strains WP2 and WP2 uvrA^– of Escherichia coli in the absence of a metabolic activation system (S9-mix). However, the valepotriates were mutagenic for TA100, WP2 and WP2 uvrA^– at concentrations up to about 1.0 mole/plate when S9-mix was added to the test system. With more than 1 mole/plate the valepotriates were toxic in the presence of a metabolic activation system for all strains tested. The mutagenicity of the valepotriates was inversely related to the protein content of the S9-mix used. The mutagenicity and toxicity of the valepotriates could be inhibited when the S9-mix was preincubated with the esterase inhibitor paraoxon (1 mM) for 5 min before the test compounds and bacteria were added. Therefore, bioactivation of the valepotriates by an enzymatic hydrolysis of their ester groups is considered. This could be proven by activating the valepotriates with purified esterase.Parts of this paper were presented at the Congress, Fortschritte in der Arzneimittelforschung, April 17–20, 1983 in Munich     

Number of cortisol time-points and dexamethasone suppression test sensitivity for major depression

Failure to suppress cortisol secretion after administration of dexamethasone has been reported to be a diagnostic marker for major depression and to have prognostic implications when repeated after antidepressant treatment. The pulsatile pattern of cortisol secretion suggested to us that increasing the number of post-dexamethasone cortisol determinations might significantly increase the sensitivity of the dexamethasone suppression test (DST) for major depression. With a conventional two-point DST (1600 h and midnight), 5% of 20 normal volunteers, 8% of 13 inpatients with non-major depressions, and 31% of 65 inpatients with primary major depression failed to suppress. With six post-dexamethasone points (0800 h, 1200 h, 1600 h, 2000 h, 2200 h, midnight), the respective percentages were 10, 15 and 44%. The additional points increased the sensitivity from 31 to 44%, mostly by identifying more major depressives with a "late escape" pattern. If a clinician is using the DST to establish a marker for major depression that can be repeated to monitor response to treatment and the likelihood of relapse, then perhaps the increased sensitivity of the six-point DST would be helpful, despite a modest decrease in specificity from 94 to 88%.     

A case of “myopathy with tubular aggregates” with increased muscle fibre sensitivity to caffeine

Summary A 23-year-old man with myopathy with tubular aggregates had suffered from exercise-induced muscle cramps for 1 year. His general and neurological findings were normal. Laboratory investigations were within normal limits except for a slightly elevated serum creatine kinase level. Muscle biopsy showed some small angular fibres and scattered type 2B fibres with prominent tubular aggregates originating from the sarcoplasmic reticulum. Since the muscle fibres contracted at a lower concentration of caffeine, increased muscle fibre sensitivity to caffeine is probably related to muscle cramps in this disorder. Tubular aggregates are then secondarily formed in the muscle fibres.     

Effects of fixed-interval duration on the development of tolerance to decreased responding byl-nantradol

The effects of several types of antidepressants in a recently developed behavioural despair model, the tail-suspension test, are described. Drug effects on the automatically recorded duration of immobility and power of movements were measured in three strains of mice. Only in one strain (NMRI) did almost all antidepressants tested showed the expected reduction in duration of immobility. Tranquillizing drugs, but not stimulants, could be distinguished from antidepressants. The power of movements could not definitively be related to the pharmacological profile of the drugs tested. The use of the tail-suspension test as a rapid and highly predictive behavioural primary screen for antidepressant drugs is suggested.     

Squat and smile assessment in predicting healing of lower limb fractures fixed with a SIGN nail

BackgroundIrrespective of the treatment method, union is the ultimate goal of any fracture treatment. However, nowadays, rather than the physician-based clinico-radiological methods, the patient-reported outcome measurements assessing their quality of life and function are gaining much popularity. This is specifically true in the part of the world where the patient needs almost complete degree of hip/knee flexion – for social, cultural, religious or occupational reason(s). The ability to squat can assess the mobility and stability of joints and thus the quality of squatting is a proxy reflection of the functional outcome after fixation of lower limb fracture. Thus, we studied to determine the inter-observer and intra-observer reliability of Radiographic Union Score for Tibia (RUST) and Squat and Smile (S & S) test in clinical photograph. We further calculated the sensitivity and specificity of S & S test in predicting healing of lower limb fracture fixed by intramedullary interlocking nail considering RUST as the gold standard.MethodsThis was a retrospective study of prospectively collected data of solid Surgical Implant Generation Network (SIGN) intramedullary interlocking nailing from a single, university-based, high volume tertiary center where 56 consecutive adults with either tibial or femoral shaft fractures fixed with a SIGN nail within one year and not requiring any surgery till minimum of eighteen-month follow-up were included. Cases without either Anterior-Posterior (AP) view and/or Lateral (Lat.) view follow-up x-ray(s) or proper S & S clinical photograph (at least 1.5-year post fixation) were excluded. The x-rays (RUST criteria) and clinical photograph (S & S grading) were scored by two independent and blinded observers each and repeated after 1 month.ResultsThe overall intra-observer reliability was from 0.773 to 0.825 and inter-observer reliability from 0.635 to 0.757 for RUST scoring which was from 0.687 to 0.785 and from 0.301 to 0.650 respectively for S & S scoring. The sensitivity and specificity of S & S in predicting fracture healing were up to 82.22% and 63.64% respectively.ConclusionThe S & S test is reliable to predict the healing of lower limb fracture fixed with an intramedullary nail. The test is more useful to determine healed fractures than to determine non-healed ones. (sensitivity being higher than specificity)     

Nerve Conduction Studies in Surgical Cubital Tunnel Syndrome Patients

Background: Electrodiagnostic studies (EDX) serve a prominent role in the diagnostic workup of cubital tunnel syndrome (CBTS), but their reported sensitivity varies widely. The goals of our study were to determine the sensitivity of EDX in a cohort of patients who responded well to surgical cubital tunnel release (CBTR), and whether the implementation of the Association of Neuromuscular and Electrodiagnostic Medicine (AANEM) criteria improves the sensitivity. Methods: We identified 118 elbows with clinical CBTS who had preoperative EDX and underwent CBTR. The EDX diagnoses were CBTS, ulnar neuropathy (UN), and normal ulnar nerves. We divided the 118 elbows into those that received above-elbow stimulation (XE group) and those that did not (non-XE group). We calculated the sensitivities for all groups and reinterpreted the results according to the AANEM guidelines. Results: Cubital tunnel release provided significant relief in 93.6% of the elbows. Based on the EDX reports, 11% patients had clear CBTS, 23% had UN, and 66% showed no UN. The sensitivities were 11.7% for CBTS and 34.2% for any UN. In the XE group, the sensitivity of the EDX reports for CBTS and UN climbed to 33.3% and 58.3%, respectively. When we calculated the across-elbow motor nerve conduction velocity, the sensitivity for CBTS and UN was 87.5% and 100%, respectively. The XE and non-XE groups showed no difference except for sex, bilaterality, concomitant carpal tunnel release, and obesity (P < .05). Conclusion: Implementing AANEM guidelines results in significant improvement in correlation of clinical and electrodiagnostic findings of CBTS.