Dual immunodetection and hybridisation in situ of opsin mRNA and rhodopsin protein in retinal sections

We describe a simple method for combining in situ hybridisation and immunohistochemistry on the same retinal section. The technique was developed using a radiolabelled cDNA probe for opsin and an antibody (ROS1F4) against rhodopsin. This method retains the antigenic sites if immunocytochemistry is performed prior to in situ hybridisation. Opsin mRNA was found in the photoreceptor inner segment with rhodopsin immunolocalised to the photoreceptor outer segments. The technique should be applicable to numerous situations including analysis of the sequence of events in the expression and synthesis of the various opsins during retinal development and degeneration.     

Prenatal determination of human platelet antigen type using DNA amplification following amniocentesis

OBJECTIVE: To assess the effectiveness of a newly developed individualised birthweight ratio (IBR), which corrects for physiological birthweight determinants, in identifying infants at risk from the complications of macrosomia. DESIGN: Prospective observational study. SETTING: Obstetric unit, Nottingham City Hospital. SUBJECTS: 2835 women delivered between December 1991 and July 1992 and the infants of 624 of these, selected by virtue of their birthweight for gestation and IBR centile positions. MAIN OUTCOME MEASURES: Skinfold thickness and ponderal index measurements, operative delivery, shoulder dystocia, fetal trauma, impaired glucose tolerance. RESULTS: Using an IBR above the 90th centile as a cut off results in 2.4% of infants being reclassified as normally grown and 3.1% are reclassified as large. The IBR does not result in the identification of any more infants with abnormal ponderal indices or skinfold thicknesses than birthweight for gestation. It does, however, identify more of the infants at risk of operative delivery, shoulder dystocia, fetal trauma and impaired glucose tolerance. CONCLUSION: The IBR significantly improves upon birthweight for gestation in identifying infants who suffer from the complications of relative macrosomia.     

Further observation of leucocyte migration through the endothelium of venules in the posterior latissimus dorsi muscle of the quail-chick spinal cord chimeras

The migration of leucocytes through the walls of venules was examined in detail in the posterior latissimus dorsi muscle of quail-chick spinal cord chimeras, in which flaccid paralysis of the wings was observed. Examinations were made from the 70th to 80th day after hatching. Muscle fibers were degenerated and intramuscular nerve bundles destroyed. Massive leucocytes (almost lymphocytes) were found around the venules, depending on the passage of wandering leucocytes through the endothelium. Lymphocytes penetrated and were encased in the cytoplasm of the venular endothelial cell, and did not pass through the interendothelial junction. These findings suggest that, in the venules of the atrophied chimeric muscle, wandering leucocytes in the blood may pass through the endothelial cell body and migrate into the inflamed extravascular space.     

Premature separation of centromere and aneuploidy: an indicator of high risk in unaffected individuals from familial breast cancer families?

OBJECTIVES: Injury is the leading cause of death in the male working population of Brazil. An important fraction of these deaths are work related. Very few cohort studies of steel workers, and none from developing countries, have reported on mortality from injuries. This paper analyses mortality from work and non-work related injuries among Brazilian steel workers. METHODS: Deaths during employment from 1 January 1977 to 30 November 1992 were analysed in a cohort of 21,816 male steel workers. Mortality rates specific for age and calendar year among the workers were compared with those of the male population of the state where the plant is located. Work related injuries were analysed by comparing the mortality rates for different subgroups of the cohort. RESULTS: The number of deaths (391) was less than half that expected based on death rates of the general population. Over 60% (242) of deaths were due to injuries. Mortality from most causes was substantially below that in the general population, but that from unintentional injury, was 50% above that of the general population. Standardised mortality ratios (SMRs) were highest for the youngest and the oldest employees and for labourers and clerical workers. Mortality from motor vehicle injury was twice that expected from population rates (SMR = 209, 95% confidence interval (95% CI) 176-244). There was a 67% fall in the age adjusted mortality from occupational injuries in the study period. CONCLUSION: The healthy worker effect in this cohort was greater than that commonly found in studies of occupational groups in developed countries, probably because of a greater socioeconomic gap between employed and unemployed populations in Brazil, and unequal distribution of health care resources. Mortality was especially high for motor vehicle injuries. The fall in mortality from occupational injuries during the study period was probably due to improvement in safety standards, increased automation, and better medical care. There is a need to investigate risk factors for unintentional injuries among steel workers, especially those due to motor vehicle injuries. Prevention of occupational and nonoccupational injuries should be a main priority in Brazil.     

Experimental occlusal interferences. Part IV. Mandibular rotations induced by a pliable interference

In 12 subjects, a pliable, yet unbreakable, intercuspal interference (aluminum shim onlay splint; uniform height of 0.25 mm) was placed between either the right or left maxillary and mandibular second premolars and first molars. During brief and forceful biting (dynamic chewing stroke of about 20 kg force) the interference emulated a semisoft food bolus, and at the end of biting (subsequent static clenching stroke of about 20 kg force) it emulated a rigid metal interference. During dynamic/static biting, rotational electrognathography measured maximum frontal and horizontal plane torque of the right and left mandibular condyles. Eleven subjects (92%) showed frontal plane upward rotation (mean of 1.0 degree) of the condyle contralateral to the interference, and one subject (8%) showed frontal plane upward rotation (0.4 degree) of the condyle ipsilateral to the interference. Two subjects (17%) showed no horizontal plane rotation; seven subjects (58%) showed backward rotation (mean of 0.4 degree) of the condyle contralateral to the interference; and three subjects (25%) showed backward rotation (mean of 0.3 degree) of the condyle ipsilateral to the interference. It is suggested that, in the presence of an occlusal interference, mastication may have both short- and long-term detrimental effects.     

CHL1 encodes a component of the low-affinity nitrate uptake system in Arabidopsis and shows cell type-specific expression in roots

The Arabidopsis CHL1 (AtNRT1) gene confers sensitivity to the herbicide chlorate and encodes a nitrate-regulated nitrate transporter. However, how CHL1 participates in nitrate uptake in plants is not yet clear. In this study, we examined the in vivo function of CHL1 with in vivo uptake measurements and in situ hybridization experiments. Under most conditions tested, the amount of nitrate uptake by a chl1 deletion mutant was found to be significantly less than that of the wild type. This uptake deficiency was reversed when a CHL1 cDNA clone driven by the cauliflower mosaic virus 35S promoter was expressed in transgenic chl1 plants. Furthermore, tissue-specific expression patterns showed that near the root tip, CHL1 mRNA is found primarily in the epidermis, but further from the root tip, the mRNA is found in the cortex or endodermis. These results are consistent with the involvement of CHL1 in nitrate uptake at different stages of root cell development. A functional analysis in Xenopus oocytes indicated that CHL1 is a low-affinity nitrate transporter with a K(m) value of approximately 8.5 mM for nitrate. This finding is consistent with the chlorate resistance phenotype of chl1 mutants. However, these results do not fit the current model of a single, constitutive component for the low-affinity uptake system. To reconcile this discrepancy and the complex uptake behavior observed, we propose a "two-gene" model for the low-affinity nitrate uptake system of Arabidopsis.