Erythroid transcription factor NF-E2 is a haematopoietic-specific basic-leucine zipper protein

Expression of globin genes in developing erythroid cells is controlled by upstream locus control regions. Activity of these regions in vivo requires an erythroid-specific nuclear factor (NF-E2) that binds AP-1-like recognition sites. Its tissue-specific component (p45 NF-E2) has been characterized by complementary DNA cloning as a new basic region-leucine zipper protein which dimerizes with a ubiquitous partner to form native NF-E2.     

Expression and possible role of muscle-type carnitine palmitoyltransferase I during sperm development in the rat

Because we had found whole testis from adult rats to be much richer in the messenger RNA for the muscle (M) than for the liver (L) form of mitochondrial carnitine palmitoyltransferase I (CPT I), we sought to determine which cell type(s) accounts for this expression pattern and how it might relate to reproductive function. Studies with immature (14-day-old) and adult animals included 1) Northern blot analysis of testis mRNA; 2) in situ hybridization with slices of testis; 3) enzyme assays for CPT I, CPT II, and carnitine acetyltransferase (CAT) in testicular germ cells and nongerm cells, together with measurement of the malonyl-coenzyme A (CoA) sensitivity and affinity for carnitine of CPT I; 4) labeling of testicular CPT I with [3H]etomoxir, a covalent inhibitor of the enzyme; and 5) the response of testicular and nontesticular CPT I to dietary etomoxir. The data established the following: 1) L-CPT I was the sole isoform detected in immature testis. 2) Expression of the M-CPT I gene was associated only with meiotic and postmeiotic germ cells. 3) Adult testis contains a mixture of the L- and M-CPT I enzymes, the L and M form dominating in extratubular cells and spermatids, respectively. Mature epididymal spermatozoa appear to be devoid of CPT I activity while possessing abundant levels of CPT II and CAT. 4) Five days of dietary etomoxir treatment at a dose that resulted in essentially complete inhibition of CPT I in liver, heart, skeletal muscle, and kidney was totally without effect on either the L- or M-type enzyme in the testis of mature rats. The data point to an important role for transient expression of M-CPT I, coupled with sustained activity of CAT, in the maturation and/or function of rat sperm. They also suggest that, at least in the case of one CPT I inhibitor (etomoxir), the testis is unusually resistant to the agent when given orally.     

Structure and function of the hepatic form of 11 beta-hydroxysteroid dehydrogenase in the squirrel monkey, an animal model of glucocorticoid resistance

Both cortisol and aldosterone bind to and activate the mineralocorticoid receptor. Cortisol concentrations are generally 100- to 200-fold higher than aldosterone concentrations, yet mineralocorticoids clearly exert effects different from glucocorticoids. One hypothesis is that 11 beta-hydroxysteroid dehydrogenase (11 beta-HSD), which converts cortisol to biologically inactive cortisone, protects the mineralocorticoid receptor from cortisol. The circulating concentrations of cortisol in the squirrel monkey are 20- to 50-fold higher than human cortisol concentrations, yet this animal has no evidence of glucocorticoid or mineralocorticoid excess. We used this experiment of nature to test the hypotheses that the known (hepatic) form of 11 beta-HSD protects renal mineralocorticoid receptors from the action of cortisol and that it modulates glucocorticoid concentrations in target tissues. Using a long oligonucleotide based on the rat sequence, we cloned the squirrel monkey 11 beta-HSD complementary DNA and gene. The encoded monkey amino acid sequence is 75% and 91% identical to the corresponding rat and human sequences, respectively. The tissue abundance of the messenger RNA for the monkey enzyme was similar to or less than that seen for the rat and human enzymes. Both the monkey and human 11 beta-HSD complementary DNAs were cloned into an expression vector and used to transfect cultures of Chinese hamster ovary cells. Both vectors were transcribed and translated into equivalent amounts of 11 beta-HSD enzyme. The monkey enzyme was slightly more efficient than the human enzyme in converting [3H]cortisol to cortisone, and estimates of the Michaelis-Menten constant and maximum velocity of both enzymes are similar. These data indicate that the abundance and activity of the hepatic form of 11 beta-HSD are insufficient to inactivate the very high concentrations of cortisol in the squirrel monkey, suggesting that this form of 11 beta-HSD does not defend the mineralocorticoid receptor or protect tissues from high cortisol concentrations. Rather, this enzyme appears to favor conversion of cortisone to cortisol, thus maximizing tissue concentrations of cortisol to overcome glucocorticoid resistance associated with a 50% reduction in glucococorticoid receptors.     

Serial light and electron microscopic changes in renal glomeruli in mice fed griseofulvin

The acclimation of the clawed toad Xenopus laevis to hyperosmotic solutions of NaCl (balanced solution of sea salt), urea or mannitol was studied. The animals could not be acclimated to salt solutions more concentrated than 400 mosm.1-1. Urea was tolerated till 500 mmol.1-1. Plasma osmolality was always hyperosmotic to the environmental solution, but with diminished osmotic gradient at the highest tolerated solutions. Plasma urea concentration approached 90 mmol.1-1, similar in the three solutions of acclimation. Urine volume was very small under all conditions. Serum aldosterone and corticosterone did not differ significantly, although there was a slight tendency towards lower aldosterone in the NaCl solution. In vivo water uptake in tap water acclimated animals was very small, and was higher in the other groups. Only the salt- and urea-acclimated, but not the tap water and mannitol-acclimated groups responded with a clear increase following injection of oxytocin or theophylline. In vitro urea fluxes were similar and invariable in both directions under all conditions. No significant effect of theophylline was observed. Sodium transport measured by the short-circuit technique in vitro was lower in salt- and mannitol-acclimation conditions, and was stimulated significantly under all conditions in response to serosal oxytocin or theophylline. It is concluded that Xenopus laevis can osmoregulate at a limited range of external solutions. It is limited in the increase of its plasma urea concentration; the transport properties of the skin do not change very much upon acclimation, except for the hydroosmotic response to oxytocin.     

Lipid Oxidation in Cooked Turkey as Affected by Added Antioxidant Enzymes

Catalase (CAT) and glutathione peroxidase (GSH-Px) were added to turkey which had been cooked (80°C) to provide minimal activity of both enzymes, to determine their effects in development of lipid oxidation. CAT (100–500 U/g muscle) decreased the formation of thiobarbituric acid reactive substances (TBARS) 4–28% during 2 days of storage. GSH-Px (4.0 U/g muscle) decreased TBARS 8–11%. CAT (170 U/g muscle) and GSH-Px (1.3 U/g muscle) in combination decreased TBARS formation 7–14%. Reduced glutathione concentrations in the turkey were unaffected by cooking. These data suggest that heat inactivation of CAT and GSH-Px was not the only factor involved in heat-induced lipid oxidation reactions in turkey thigh muscle.     

Quantitative contribution of the collagen network in dentin hybridization

PURPOSE: To determine the quantitative contribution of dentin hybridization to bonded assembly strength and demonstrate the micromorphology of the interface with and without collagen present. MATERIALS AND METHODS: Four groups of 10 molar teeth were finished to a 320 grit dentin smear layer. Two groups served as controls and two experimental groups were subjected to collagenase digestion of the collagen exposed by acid conditioning. All-Bond 2 and Amalgambond were used to bond Bisfil and Epic resin composite, respectively. Stored in water at 37 degrees C for 24 hours the assemblies were tested in a shear mode at a crosshead speed of 5 mm/minute. Means and standard deviations were subjected to analysis for statistical significance. Twenty four teeth in four groups were examined by scanning (SEM) and transmission electron microscopy (TEM) for the relationship between resin and conditioned dentin with and without the collagen network. RESULTS: All-Bond 2 and Amalgambond controls were 28.41 +/- 3.9 and 19.04 +/- 5.96 MPa, collagenase-treated groups scored 26.43 +/- 2.90 and 19.70 +/- 4.25 MPa respectively. No significant difference existed between the control and experimental groups. SEM showed an intertubular collagen network with patent tubules and a pronounced porous, irregular dentin topography following collagen digestion. A distinct hybrid zone and tubular penetration was observed but the collagenase-treated specimens showed only resin in the tubules and their lateral extensions. TEM confirmed the absence of a distinct hybrid zone in the collagenase groups with a tight, gap-free junction between the resin and the undemineralized dentin. An electron dense zone (< 50 nm) at the leading edge of conditioning was observed for All-Bond 2 and Amalgambond groups. It was concluded that the resin-reinforced or hybridized, collagenous network does not detract from, nor contribute any significant quantitative value per se to dentin bonding with the systems tested.     

Prediction of facial nerve displacement in extra large vestibular schwannoma

The primary objective in the surgery of extra large vestibular schwannoma is the total removal of the tumour mass while preserving the facial nerve. Preservation of the facial nerve in extra large tumours is reported as being notoriously difficult in the majority of cases. This study was undertaken to evaluate the accuracy in predicting displacement of the facial nerve by preoperative radiological imaging studies in 19 cases of extra large vestibular schwannoma. The direction of displacement of the facial nerve was predicted with preoperative axial and coronal MRI scans and verified intraoperatively. We achieved total removal of tumours in 84.2%, facial nerve displacement was predicted in 80% and we accomplished anatomical preservation in 80%. Prediction of displacement was difficult in tumours with little or no intracanalicular components or with severe bony destruction of the internal acoustic meatus. The preoperative prediction of facial nerve displacement in extra large tumours allows safe internal decompression of the tumour and careful dissection near the predicted area of the facial nerve during the operation. Consequently, a high rate of anatomical preservation of the facial nerve can be achieved.     

Early IL-4 induction in bone marrow lymphoid precursor cells by mycobacterial lipoarabinomannan

IL-4 is produced promptly in response to certain infections and plays a key role in the Th1/Th2 T cell dichotomy; however, the cellular source remains a matter of debate. Here we describe the induction of IL-4 in bone marrow cells of normal and RAG-/- mice by both Mycobacterium tuberculosis and its major cell wall glycolipid, lipoarabinomannan. Characterization of the cell type responsible indicated that it was distinct from the NK1+ or CD4+ T cell previously ascribed the function of rapid IL-4 secretion. Cell-sorting experiments identified CD19+/B220+ precursor cells, presumably pre-B cells that produced IL-4 constitutively and whose frequency was rapidly and markedly up-regulated by lipoarabinomannan. Thus, pathogenic mycobacteria and their glycolipids may influence hemopoiesis by rapidly inducing IL-4 secretion in the bone marrow.     

Lumbar strengthening in chronic low back pain patients. Physiologic and psychological benefits

The effects of exercise for isolated lumbar extensor muscles were examined in 54 chronic low-back pain patients. Subjects were randomly assigned to a 10-week exercise program (N = 31) or a wait-list control group (N = 23). Results indicated a significant increase in isometric lumbar extension strength for the treatment group and a significant reduction in reported pain compared with the control group (P 0.05). Treated subjects reported less physical and psychosocial dysfunction whereas the control group increased in pain, and physical and psychosocial dysfunction. There were no concomitant changes in reported daily activity levels. These results show that lumbar extension exercise is beneficial for strengthening the lumbar extensors and results in decreased pain and improved perceptions of physical and psychosocial functioning in chronic back pain patients. However, these improvements were not related to changes in activities or psychological distress.