The potential for functional recovery of upper extremity function following cervical spinal cord injury without major bone injury.

STUDY DESIGN: This was a retrospective observational study. OBJECTIVES: The objectives were to describe the prognosis of upper extremity function following cervical spinal cord injury (CSCI), and to identify prognostic factors for functional recovery. SETTING: Spinal Injuries Center, Japan. METHODS: Sixty patients with C3-4 CSCI without major bone injury participated in the study. Patients were treated nonsurgically and evaluated using the American Spinal Injury Association (ASIA) scales for the upper and lower extremities, their residual cervical motor functions, the modified Frankel grade and an upper extremity function scale. We compared the findings for the upper extremity function scale at 6 months with those for the residual cervical motor functions and modified Frankel grade obtained 3 days after injury. RESULTS: Most patients with CSCI who could flex their hip and knee from a supine position (95%) or who showed some active elbow extension (86%) 3 days after their injury could use a spoon at 6 months. We compared patients who used their fingers at 6 months to those who could not, and observed significant differences in age and ASIA scores for the upper and lower extremities obtained 3 days after injury. A strong correlation was observed between the initial motor scores and the extent of functional recovery at 6 months. CONCLUSION: Hip and knee flexion from the supine position and elbow extension 3 days after injury significantly predicted a positive prognosis for upper extremity function. Younger age and higher ASIA motor scores obtained 3 days after injury were factors associated with neurological recovery.

Isolation and characterization of cDNA for an androgen-regulated mRNA in the flank organ of hamsters.

Flank organs of hamster are useful for studying androgen-dependent growth of hair follicles and sebaceous glands. To elucidate the mechanism of gene expression regulated by androgen, we constructed a cDNA library from flank organs of male hamsters and screened by a differential hybridization method using cDNA probes from normal and castrated males. We isolated a cDNA clone, termed FAR-17a, whose expression was found to be highly sensitive to androgen. FAR-17a mRNA of 1.8 kb was reduced after castration and reappeared after testosterone treatments. Among several tissues examined, FAR-17a gene was expressed at a high level in flank organ and a low level in testis and earlobe. FAR-17a probe detected a few fragments in genomic DNA of hamster, mouse, suncus, pig, and human, suggesting that this gene is phylogenetically conserved. The sequence of FAR-17a cDNA predicts a protein of 231 amino acids (27,216 daltons) having basic properties. The deduced protein has no significant homologies to proteins previously described.

Sequence analysis and characterization of FAR-17c, an androgen-dependent gene in the flank organs of hamsters.

This study reports on the isolation and characterization of a cDNA clone, regulated by androgen transcriptionally, in male Golden hamsters' flank organs. Previous studies have reported on the cloning of an androgen-dependent gene, FAR-17a, from the same hamster organ. After castration, the FAR-17c transcription rate decreases faster than FAR-17a but is not suppressed completely. The recovery of transcription by androgen injection is also faster than FAR-17a. In male hamsters, it is expressed strongly in the sebaceous glands and liver, and weakly in the lungs and brain. It has never been expressed in the testes. In the female, it is strongly expressed in the liver and brain and weakly in the lungs and flank organs. Sequence analysis shows that FAR-17c has a long 1062 bp open reading frame and its deduced amino acid sequence (354 residues) is highly homologous to the stearyl-CoA desaturases of the rat liver and mouse adipocytes. Stearyl-CoA desaturase, either in the liver or adipocytes appears to be independent of androgen regulation. Since stearyl-CoA desaturase plays a key role in fatty acid metabolism, further studies on its regulation by androgen are warranted in relation to acne vulgaris.

Molecular cloning and DNA sequence of dniR, a gene affecting anaerobic expression of the Escherichia coli hexaheme nitrite reductase.

A gene responsible for increased synthesis of hexaheme nitrite reductase (cytochrome c552) of Escherichia coli K-12 was cloned into pBR322 by the direct immunological screening method using antiserum against the purified enzyme. The cloned gene was mapped at 5 min on the chromosomal linkage map as the dni gene (related to increased synthesis of the dissimilatory nitrite reductase) by conjugation and transduction. The dni gene was subcloned into pUC118 and was shown to be on a 2.6-kilobase-pair PstI-BamHI fragment by immunoblotting analysis of the expressed enzyme. The nucleotide sequence of this fragment was determined. A plausible open-reading frame corresponding to 222 amino acids was detected. Analysis of a dni deletion mutant by immunoblotting demonstrated that this mutant expressed a greatly reduced amount of the nitrite reductase. Thus, the dni gene is suggested to have a positive regulatory action on induced synthesis of the nitrite reductase, and was designated as dniR.

Hyaluronan synthase in trabecular meshwork cells.

BACKGROUND/AIMS: Hyaluronan is present in the trabecular meshwork where it is involved in the pathophysiology of aqueous outflow environment. In this study, the expression and regulation of hyaluronan synthase (HAS), which is the enzyme synthesising hyaluronan, in trabecular meshwork cells were investigated. METHODS: Cultured bovine trabecular meshwork cells (BTMCs) were used. HAS expression in BTMCs was examined by RT-PCR. The effects of transforming growth factor beta (TGF-beta) and platelet derived growth factor BB (PDGF-BB) on HAS expression in BTMCs were examined by quantitative RT-PCR. The HAS2 expression by TGF-beta and PDGF-BB at the protein level was also confirmed immunohistochemically. The production of hyaluronan from BTMCs was detected by high performance liquid chromatography (HPLC). RESULTS: Three HAS isoforms were expressed in BTMCs at the mRNA level. Among HAS isoforms, only the expression of HAS2 mRNA was increased by the administration of TGF-beta or PDGF-BB. HAS2 upregulation by these growth factors was also confirmed at the protein level. Further, hyaluronan production from BTMCs was stimulated by TGF-beta or PDGF-BB. CONCLUSION: Expression of HAS in trabecular meshwork may maintain the hyaluronan content in the aqueous outflow pathway. Its production is regulated by TGF-beta and PDGF-BB. The regulation of the expression of HAS in trabecular meshwork might be useful for modulating the aqueous outflow environment.

Androgen regulation of a specific gene in hamster flank organs.

Flank organs of Golden Syrian hamster containing large sebaceous glands are a useful model for studying androgen-dependent gene expression. Recently, we have isolated an androgen-dependent gene from a cDNA library constructed from male mRNAs of flank organs. In the present study, using this gene as a probe, we examined the kinetics of loss and re-expression of the mRNA of this gene. Dot blot hybridization, the current method, can approximately quantify mRNA levels. Castration caused a marked decrease in the mRNA within a few days to undetectable levels. Topical application of testosterone re-activated the mRNA levels, with the earliest activation within 24 h. The results of various topical steroid applications showed a full re-activation of the mRNA by testosterone and dihydrotestosterone, partial re-activation by androstanedione and androstenedione, and no activation by androstanediol or dehydroepiandrosterone. Simultaneous application of testosterone and progesterone inhibited the expression of mRNA but application of dihydrotestosterone and progesterone did not. The current dot blot hybridization technique appears to be a more direct approach for studying androgen action on DNAs than other previously used methods such as morphometry or enzyme assay.

Central corneal thickness of normal tension glaucoma patients in Japan.

PURPOSE: To compare central corneal thickness (CCT) of patients with normal tension glaucoma (NTG) with that of age-matched normal subjects, patients with open-angle glaucoma (POAG) and ocular hypertension (OH) subjects in Japan. METHODS: Central corneal thickness was measured in 79 NTG, 61 POAG, 73 OH, and 50 normal subjects with an ultrasonic pachymeter. One eye for 1 subject randomly selected in each group was used for inter-group comparison. The relationship between CCT and the maximum intraocular pressure (IOP) measured by Goldmann applanation tonometer with no ocular hypotensive medication (NTG, OH, and normal subjects) or under medication (POAG patients) was analyzed. RESULTS: The CCT of OH subjects (582 +/- 32 microm; mean +/- SD) was significantly greater than that of the other groups (P <.001), while no difference was seen in CCT among normal (552 +/- 36 microm), NTG (548 +/- 33 microm) and POAG (550 +/- 33 microm) subjects. In normal subjects, CCT and the maximum IOP were significantly correlated but the correlation coefficient was small (r = 0.420, P <.05). CONCLUSIONS: Central corneal thickness shows no significant difference among NTG, POAG, and normal subjects in Japan, while it is significantly greater in OH subjects. The CCT has little influence on the diagnosis of NTG in Japan.

Characterization of the promoter region of the human melanocortin-1 receptor (MC1R) gene.

We sequenced 3201 bp upstream from the ATG translation start codon of the human melanocortin-1 receptor (MC1R). A number of transcriptional initiation sites were detected over a region of approximately 600 base pairs upstream of the receptor coding region. These consist of GC-rich regions, each including SP-1 consensus binding motifs. Neither a TATA nor a CAAT box was found in this region. The 5'-flanking region also contains the consensus regulatory elements for AP-1, AP-2, and several E-boxes. Gel shift assays targeting the three GC boxes confirmed binding of SP-1. A promoter assay revealed that the minimal region exhibiting promoter activity was located between nucleotides -517 and -282 in human melanoma SK-Mel-2 cells. Further deletion from -517 to -447, which removed an SP-1 site, completely abolished luciferase activity. In conclusion, the MC1R promoter shares the characteristics of many other GPCR promoters. These characteristics include GC-rich sequence, lack of a TATA box, and binding of SP-1.

The isolation and characterization of androgen-dependent genes in the flank organs of golden Syrian hamsters.

To elucidate the molecular action of androgens, we have isolated two androgen-dependent genes, FAR-17a and -17c, from the cDNA library of flank organs of male golden Syrian hamsters by a differential hybridization method. FAR-17a has been reported previously. Androgens regulated the expression in the flank organs, ear lobe and skin. FAR-17a protein was located in sebaceous glands. FAR-17c has a similar expression pattern to that of FAR-17a, but its response to androgen is faster than that of FAR-17a. It is expressed strongly in the liver, sebaceous glands and brain. The homology search and mRNA expression pattern suggested that it might encode a stearyl CoA desaturase (SCD) of golden Syrian hamsters. The SCD is a key enzyme in fatty acid biosynthesis and its expression is controlled by temperature, nutritional conditions and hormones. This is the first report that the mRNA expression of stearyl CoA is regulated by androgens.

Human and rat TR4 orphan receptors specify a subclass of the steroid receptor superfamily.

We have identified a member of the steroid receptor superfamily and cloned it from human and rat hypothalamus, prostate, and testis cDNA libraries. The open reading frame between first ATG and terminator TGA can encode 615 (human) and 596 (rat) amino acids with calculated molecular mass of 67.3 (human) and 65.4 (rat) kDa. The amino acid sequence of this protein, called TR4 orphan receptor, is closely related to the previously identified TR2 orphan receptor. The high homology between TR2 and TR4 orphan receptor suggests that these two orphan receptors constitute a unique subfamily within the steroid receptor superfamily. These two orphan receptors are differentially expressed in rat tissues. Unlike TR2 orphan receptors, the TR4 orphan receptor appears to be predominantly located in granule cells of the hippocampus and the cerebellum, suggesting that it may play some role(s) in transcriptional regulation in these neurons.

Gene expression of the androgen repressed rat TR2 orphan receptor: a member of steroid receptor superfamily.

A full-length rat cDNA clone was obtained from the TR2 orphan receptor, a member of the steroid receptor superfamily, using cDNA library screening and 3' RACE-PCR technology. Under these conditions, only the TR2-11 form of the TR2 orphan receptor, the major form found in prostate, was identified. The overall amino acid homology between human and rat TR2-11 orphan receptors was near 90% with one amino acid difference in the DNA-binding domain sequence. Northern blot analysis identified multiple forms of the TR2 orphan receptor mRNAs expressed in human and rat prostates. Androgens repressed TR2 orphan receptor mRNA levels in human prostate LNCaP cells and rat ventral prostate. Polyclonal anti-TR2 orphan receptor antibodies raised from a unique TR2 orphan receptor 20 amino acid peptide were used to localize the TR2 orphan receptor in the nuclei of prostate and epididymis epithelium cells. Together, these data demonstrate that the TR2 orphan receptor can be expressed at mRNA and protein levels in the human and rat prostrates and may have some potential function in mediating androgen action in these tissues.