Absence of interferons-alpha and -gamma in renal lesions of systemic lupus erythematosus and membranous glomerulonephritis

Frozen kidney biopsy sections from nine patients with systemic lupus erythematosus (SLE) as well as many other renal diseases, including IgA nephropathy, membranous nephritis, and minimal change nephrotic syndrome, were negative for interferons -alpha and -gamma by immunofluorescence. Lupus patients studied included several subjects with marked serum elevations of interferon activity as well as others with low or negative serum interferon levels. Isolated glomerular eluates prepared from normal and SLE kidneys showed no functional interferon activity by virus plaque inhibition assay. Components of normal as well as SLE serum showed no direct binding to interferon -alpha or -gamma by ELISA assays.     

Clonidine suppression and its adrenoreceptor mediation in schedule-induced polydipsia

Rats were trained in a fixed-interval, one-minute (FI 1 min) food reinforcement schedule for 1 hour daily at reduced body weight until their lever presses, licks and water intake all became stabilized for 6 days. Two experiments were performed to examine the function of sympathetic activity in schedule-induced polydipsia. In experiment 1, intracerebroventricular injection of clonidine (0.75-37.5 nmol) produced a dose-related suppression of schedule-induced drinking and licking and schedule-dependent lever pressing; these effects were later attenuated by yohimbine (5 nmol) pretreatment. Prazosin (10 nmol) also decreased clonidine-induced suppression of lever pressing, whereas neither prazosin (10 nmol) nor naloxone (10 nmol) caused any alteration in the suppression effects of clonidine on drinking and licking. None of these antagonists alone changed an individual rat's preestablished behavioral baselines. In experiment 2, the endogenous catecholamine levels, were determined in frontal cortex, hypothalamus, brainstem, dorsal obex area and adrenal glands. During the SIP situation, both the epinephrine level in adrenal glands and the norepinephrine level in hypothalamus were elevated.     

Biochemical characterization of human Thy-1

The human Thy-1 homologue (p25) was characterized biochemically for amino acid composition, sequence and carbohydrate content. Two other forms of the human Thy-1 molecule were detected and partially characterized. A 40,000 mol.wt. molecule (p40) is the dimer of p25 and its formation is increased by the presence of sodium dodecyl sulphate (SDS). The second form of 16,000 mol.wt. (p16) appears to be a cryptic or breakdown form of p25. Comparison of the amino acid compositions of p25, p40 and p16 isolated from MOLT-3 cells, with that deduced from the nucleotide sequence of the gene coding for part of the putative T cell antigen receptor, also from MOLT-3 cells, shows that the Thy-1 homologue is distinct from, but evolutionary related to, one of the putative T cell antigen receptor polypeptide chains.     

Topography and behavior of Sertoli cells in sparse culture during the transitional remodeling phase

We report observations on the behavior of Sertoli cells in sparse culture during the period from the time of plating to the time of initial confluence (the transitional remodeling phase). Changes in shape, structure, and polarity of cells, as well as changes in migration patterns and cell-cell association patterns, have been followed during the transitional remodeling phase with the aid of topographical markers. These markers are based upon differences between ultrastructural features of the basolateral and apicolateral surfaces. The basolateral surface is characterized by plasmalemmal blebs, whereas the apicolateral surface is characterized by filopodial extensions. Structural differences observed in situ remain evident in Sertoli cells isolated by sequential enzymatic treatments that are described. Another marker is provided by laminin-binding sites, which are detected exclusively on the blebbed, basolateral surfaces of freshly prepared Sertoli cell aggregates. The orientation described is sustained during the initial radial migration of Sertoli cells explanted on uncoated glass coverslips. Under these conditions, blebs are detected only on the dorsal surfaces, and filopodial extensions are evident only on the ventral surfaces. In contrast, Sertoli cells sparsely plated on a reconstituted basement membrane (air-dried Matrigel) migrate rapidly, display an extraordinary capacity to form elaborate cytoplasmic extensions for cell-cell and cell-substratum contacts, and readily retract blebs and filopodial extensions. These cells do not form mosaic borders, whereas cells plated on uncoated glass do form a monolayer with mosaic-like borders. Cells sparsely seeded on gelated Matrigel migrate preferentially at gaps between adjacent cell explants, and develop a compact cell-cell association pattern. These cells display few, if any, cytoplasmic extensions. We compare the behavior of Sertoli cells sparsely plated on Matrigel with the behavior of Sertoli cells in situ during different stages of development.     

Effect of hypothermia on the intestinal absorption of uracil and L-dopa in the rat

The in situ rat gut technique was used to explore the effects of hypothermia on the intestinal absorption of L-dopa and uracil. A hypothermic state was induced when male Sprague-Dawley rats, weighing between 300 and 370 g, were exposed to an atmosphere of helox (helium:oxygen, 80:20) at 0-4 degrees C. After 4 to 5 h, the rectal temperatures were decreased from 37 to 20 degrees C. The rewarming process for hypothermic animals undergoing anesthesia appears to be prolonged. The animals were prepared for surgery using ether as anesthetic after a rectal temperature of 20 degrees C was attained. Hypothermia showed a significant influence on decreasing L-dopa and uracil disappearances from the intestinal lumen. About 40 percent reduction of the rates of disappearance was observed with a 10 degrees C reduction of the rectal temperature. Additionally, a rapid drop of the rate of drug disappearance was observed in the beginning stage of rectal temperature decrease (31-36 degrees C) as well as in the hypothermic state with rectal temperatures < 26 degrees C. There appears to be a thermal stable region for absorption between 26 and 30 degrees C. Water efflux was studied in both normothermic and hypothermic animals. Normothermic rats showed greater average cumulative water efflux over a period of 30 min (71 and 61% for L-dopa and uracil, respectively) in comparison with that of hypothermic rats (34 and 41% for L-dopa and uracil, respectively). The cold-treated animals showed decreased rates of disappearance associated with the decreased water efflux. Therefore, the reduction of the rates of drug disappearance from the intestinal lumen caused by hypothermia may be partially related to the decrease of water efflux during hypothermia.     

Immunogenicity,safety and reactogenicity of ROTAVAC® in healthy infants aged 6–8 weeks in Vietnam

BackgroundROTAVAC® is derived from human 116E rotavirus (RV) neonatal strain. In this study, we evaluated the immunogenicity, safety and reactogenicity of ROTAVAC® in Vietnam.MethodWe conducted a phase IV clinical trial in healthy infants aged 6–8 weeks using the complete regimen of ROTAVAC® with three doses. Serum anti-RV IgA was measured by enzyme-linked immunosorbent assay to assess the geometric mean concentration in infants who received the complete regimen of the vaccine.ResultsA total of 360 participants were enrolled in this clinical trial. The mean age ± standard deviation at enrollment was 6.9 ± 0.6 weeks. The anti-RV IgA titer was 4.01 ± 3.74 mg/ml pre-vaccination and substantially increased to 29.27 ± 80.64 mg/ml post-vaccination. The value of logIgA significantly increased (p = 0.003) from 0.28 ± 0.79 to 1.03 ± 0.54. The proportion of participants with equal to and greater than 3-fold and 4-fold shifts in pre- to post-vaccination antibody titer (IgA) were 55.4% and 48.3%, respectively. No adverse events or serious adverse events were recorded immediately within 30 min after the administration of each dose. The most common adverse events within 14 days after each visit were fever, unusual crying and irritability. Other adverse events occurred at a low rate, and no case of intussusception was noted.ConclusionsThe complete regimen of ROTAVAC® demonstrated an immunological response with clinically acceptable safety profile. Post-completion of this study, ROTAVAC® is now a WHO-prequalified vaccine and available in Vietnam.     

D-amphetamine-induced depletion of energy and dopamine in the rat striatum is attenuated by nicotinamide pretreatment

The present study examined the effects of nicotinamide on the D-amphetamine (AMPH)-induced dopamine (DA) depletion and energy metabolism change in the rat striatum. In chronic studies, co-administration of AMPH with desipramine, a drug that retards the metabolism of AMPH, (10 mg/kg, intraperitoneal [i.p.], respectively) caused a significant decrease of striatal DA content measured 7 days later. Pretreatment with nicotinamide (500 mg/kg, i.p.), the precursor molecule for the electron carrier molecule nicotinamide adenine dinucleotide (NAD), attenuated this effect of AMPH, whereas itself exerted no long-term effect on striatal DA content. In acute studies, a decrease in striatal adenosine triphospate/adenosine diphosphate (ATP/ADP) ratio was found 3 h after co-injection of AMPH and desipramine. However, nicotinamide pretreatment blocked the reduced striatal ATP/ADP ratio and resulted in a striking increase in striatal NAD content in AMPH-treated rats. Furthermore, nicotinamide was noted to increase striatal ATP/ADP ratio and NAD content in saline-treated rats. These findings suggest that nicotinamide protects against AMPH-induced DAergic neurotoxicity in the striatum of rats via energy supplement.     

Interobserver agreement for grating acuity and letter acuity assessment in 1- to 5.5-year-olds with severe retinopathy of prematurity.

PURPOSE: To evaluate interobserver test-retest reliability of the Teller Acuity Card procedure for assessment of grating acuity at ages 1, 2, 3.5, 4.5, and 5.5 years, for HOTV letter acuity at 3.5 and 4.5 years, and for Early-Treatment Diabetic Retinopathy Study (ETDRS) letter acuity at 5.5 years in the multicenter study of Cryotherapy for Retinopathy of Prematurity (CRYO-ROP). METHODS: Subjects were the 73 participants in the CRYO-ROP study who had acuity assessed at one or more ages by two of the seven study visual acuity testers as part of a quality control procedure. All subjects had birth weights of less than 1251 g, and all had severe (threshold) ROP in one or both eyes. RESULTS: For sighted eyes, interobserver agreement for grating acuity (across all five test ages) was 0.5 octave or better in 57% of eyes and 1.0 octave or better in 85% of eyes. Interobserver agreement for letter acuity (3.5-, 4.5-, and 5.5-year test ages) was 0.5 octave or better in 71% of eyes and 1.0 octave or better in 93% of eyes. For all eyes (sighted and blind), Kendall rank correlation coefficients (Tau) were 0.86, 0.83, and 0.94 for grating, HOTV, and ETDRS acuity, respectively. Kappa statistics on data from all eyes indicated excellent interobserver agreement for grating, HOTV, and ETDRS acuity (0.73, 0.80, and 0.84, respectively). Interobserver agreement was not related to age or to severity of retinal residua of ROP. CONCLUSIONS: Excellent interobserver agreement for grating acuity measurements and for letter acuity measurements was obtained. Results suggest that with careful training and implementation of quality control procedures, high reliability of visual acuity results is possible in clinical populations of young children.