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1.
Am J Pathol ; 156(5): 1685-92, 2000 May.
Article in English | MEDLINE | ID: mdl-10793079

ABSTRACT

Physiological scrotal hypothermia is necessary for normal spermatogenesis and fertility in mammals. Human RNA binding motif protein 3 (RBM3) is structurally highly similar to the cold-inducible RNA-binding protein (Cirp), and both mRNAs are induced in human cells at the scrotal temperature (32 degrees C). We report here the cloning of mouse Rbm3 cDNA, which encoded an 18-kd protein with 94% identity in amino acid sequence to that of human RBM3. In the testis of adult mice, Rbm3 mRNA and protein were detected in Sertoli cells, but not germ cells, of seminiferous tubules at all stages. The expression was not observed in Sertoli cells of fetuses, but was observed in newborn and older mice. In the TAMA26 mouse Sertoli cell line, the Rbm3 expression level was increased or decreased within 12 hours after temperature shift from 37 degrees C to 32 degrees C or 39 degrees C, respectively. In contrast to Cirp, the cold-induced growth suppression of TAMA26 cells was not affected by suppression of the Rbm3 expression. When mouse testis was exposed to heat stress by experimental cryptorchidism, the level of Rbm3 was decreased in Sertoli cells. Rbm3 may play important roles distinct from those played by Cirp in spermatogenesis and cryptorchidism by regulating the gene expression in Sertoli cells.


Subject(s)
Cryptorchidism/genetics , RNA-Binding Proteins/genetics , Sertoli Cells/metabolism , Testis/metabolism , Amino Acid Sequence , Animals , Base Sequence , Blotting, Northern , Cell Line , Cells, Cultured , Cloning, Molecular , Cryptorchidism/pathology , DNA, Complementary/chemistry , DNA, Complementary/genetics , Female , Gene Expression Regulation , Gene Expression Regulation, Developmental , Male , Mice , Mice, Inbred C57BL , Mice, Mutant Strains , Molecular Sequence Data , RNA, Messenger/genetics , RNA, Messenger/metabolism , Sequence Analysis, DNA , Sequence Homology, Amino Acid , Sertoli Cells/cytology , Temperature , Testis/cytology , Testis/growth & development , Tissue Distribution
2.
Hum Gene Ther ; 10(11): 1819-31, 1999 Jul 20.
Article in English | MEDLINE | ID: mdl-10446922

ABSTRACT

Using murine spermatogenic cell lines GC-1 spg and GC-2 spd(ts) as target cells, an attempt was made to design a retroviral vector that would transduce genes efficiently. Promoter activities of various retroviral long terminal repeats (LTRs) were examined by using chloramphenicol acetyltransferase (CAT) as a reporter. The U3 region of spleen focus-forming virus (SFFVp) showed higher enhancer activity than that of Moloney murine leukemia virus (Mo-MuLV) in both cell lines. The U3 region of myeloproliferative sarcoma virus (MPSV) showed higher activity only in GC-1 spg cells. Expression was suppressed by the repressor element of the primer-binding site (PBS) of the Moloney-related virus. The efficiency of transduction of the multidrug-resistance gene (mdr-1) by an Mo-MuLV-based vector was compared with hybrid vectors consisting of the murine embryonic stem cell virus (MESV) PBS and the LTR of either SFFVp or MPSV. Rhodamine efflux assays and colchicine-resistant colony-forming assays demonstrated higher gene expression by the hybrid vectors. Amphotropic and ecotropic receptors were found to be expressed and functional in both cell lines. Thus, these hybrid vectors represent a powerful tool by which to transfer genes into spermatogenic cells.


Subject(s)
Gene Transfer Techniques , Genetic Vectors , Retroviridae/genetics , Spermatogenesis , Spermatozoa , Animals , Cell Line , Chloramphenicol O-Acetyltransferase/metabolism , Colchicine/pharmacology , Flow Cytometry , Gene Expression , Genes, MDR , Genes, Reporter , Male , Mice , Plasmids , Rhodamines/metabolism , Transduction, Genetic
3.
Am J Pathol ; 152(1): 289-96, 1998 Jan.
Article in English | MEDLINE | ID: mdl-9422546

ABSTRACT

Physiological scrotal hypothermia is necessary for normal spermatogenesis and fertility in mammals. Cirp is a recently identified cold-inducible RNA-binding protein that is inducible at 32 degrees C in mouse somatic cells in vitro. Cirp is constitutively expressed in the testis of mouse and structurally highly similar to RBM1, a candidate for the human azoospermia factor. To elucidate the role played by Cirp in spermatogenesis, we investigated its expression levels during spermatogenesis and after heat stress. In the mouse testis, cirp mRNA was detected in the germ cells, and the level varied depending on the stage of differentiation. Also, a high level of Cirp protein was detected immunohistochemically in the nucleus of primary spermatocytes. Expression of Cirp was decreased in the GC-2spd(ts) mouse germ cell line when culture temperature was raised from 32 degrees C to 37 degrees C. When mouse testis was exposed to heat stress by experimental cryptorchidism or immersion of the lower abdomen in warm (42 degrees C) water, the expression of Cirp was decreased in the testis within 6 hours after either treatment. In human testis with varicocele analyzed immunohistochemically, germ cells expressed less Cirp protein than those in the testis without varicocele. These results demonstrated that CIRP expression is down-regulated at elevated temperature in male germ cells of mice and humans. Analysis of Cirp expression in the testes will help elucidate the molecular mechanisms leading to male infertility.


Subject(s)
Germ Cells/metabolism , Hot Temperature , RNA-Binding Proteins/metabolism , Stress, Physiological/metabolism , Testis/metabolism , Adult , Animals , Cold Temperature , Cryptorchidism/metabolism , Humans , Immunohistochemistry , In Situ Hybridization , Male , Mice , Mice, Mutant Strains , Reference Values , Spermatocytes/metabolism , Testis/pathology , Varicocele/metabolism , Varicocele/pathology
4.
Biochem Biophys Res Commun ; 236(3): 804-7, 1997 Jul 30.
Article in English | MEDLINE | ID: mdl-9245737

ABSTRACT

Although the cold-shock responses of microorganisms have been extensively investigated, those of mammalian cells are just beginning to be understood. Recently, CIRP, a member of the glycine-rich RNA-binding protein (GRP) family, has been identified as the first cold-shock protein in mammalian cells. Here, we report that RBM3, another member of the GRP family, is induced in human cells in response to cold stress (32 degrees C). RBM3 transcripts were constitutively expressed in all cell lines examined including K562, HepG2, NC65, HeLa, and T24 cells. In all of them, the transcript levels of RBM3 were increased at 24 h after the 37 to 32 degrees C temperature down-shift. In NC65 cells, the kinetics of RBM3 induction was different from that of CIRP. Protein synthesis inhibitors cycloheximide and puromycin induced RBM3 transcripts, but cadmium chloride, H2O2, ethanol, and osmotic shock had no effect. Combined with the different tissue distribution of expression, these results suggest that RBM3 and CIRP play distinct roles in cold responses of human cells.


Subject(s)
Cold Temperature , RNA, Messenger/biosynthesis , RNA-Binding Proteins/genetics , Carcinoma, Hepatocellular/metabolism , Cycloheximide/pharmacology , HeLa Cells/metabolism , Humans , Kidney Neoplasms/metabolism , Kinetics , Leukemia/metabolism , Liver Neoplasms/metabolism , Protein Synthesis Inhibitors/pharmacology , Puromycin/pharmacology , RNA, Messenger/analysis , Tissue Distribution , Tumor Cells, Cultured , Urinary Bladder Neoplasms/metabolism
5.
Hinyokika Kiyo ; 43(3): 241-4, 1997 Mar.
Article in Japanese | MEDLINE | ID: mdl-9127764

ABSTRACT

The patients, diseases and operations experienced between 1975 and 1994 in our department were statistically analyzed. The numbers of in-patients and operations have been increasing since 1977. During these 20 years, endoscopic surgery has replaced many open surgical procedures. The introduction of extracorporeal shock wave lithotripsy has dramatically changed the therapeutic modality for urolithiasis, and decreased of the necessity of open surgery.


Subject(s)
Hospitalization/statistics & numerical data , Urologic Diseases/surgery , Urologic Neoplasms/surgery , Urology Department, Hospital/statistics & numerical data , Female , Hospitals, University , Humans , Japan/epidemiology , Male
6.
Hinyokika Kiyo ; 43(1): 61-3, 1997 Jan.
Article in Japanese | MEDLINE | ID: mdl-9046426

ABSTRACT

Neoplasms metastasizing to the penis are uncommon, and occur at the end stage of carcinoma. We describe two patients with a metastatic lesion to the penis from lung squamous cell carcinoma and pancreatic adenocarcinoma. We reviewed 101 cases with secondary penile tumor. The prognosis of secondary penile tumor is extremely poor.


Subject(s)
Adenocarcinoma/secondary , Carcinoma, Squamous Cell/secondary , Lung Neoplasms/pathology , Pancreatic Neoplasms/pathology , Penile Neoplasms/secondary , Aged , Humans , Male , Middle Aged , Prognosis
7.
Gene ; 204(1-2): 115-20, 1997 Dec 19.
Article in English | MEDLINE | ID: mdl-9434172

ABSTRACT

Cold stress induces in microorganisms the synthesis of several proteins that are involved in various cellular processes such as transcription, translation and recombination. Recently, the cold-inducible RNA-binding protein (Cirp) was found to be induced in rodent cells by mild cold stress (32 degrees C). Cirp consists of an N-terminal RNA-binding domain and a C-terminal Gly-rich domain, and plays an essential role in cold-induced suppression of cell proliferation. We report here the cloning of a cDNA encoding an 18-kDa protein with 95.3% identity in an amino-acid sequence to that of mouse Cirp. The human CIRP gene has been mapped to the chromosomal locus 19p13.3 by fluorescence in-situ hybridization. CIRP mRNA is constitutively expressed in all cell lines examined, including K562, HepG2, NC65, HeLa, T24, and NEC8 cells. In all of them, the levels of CIRP mRNA and protein were increased within 12 h after a temperature down-shift from 37 degrees C to 32 degrees C. These results demonstrated that CIRP is a cold-shock protein in human cells. Identification of CIRP may provide a clue to the regulatory mechanisms of cold responses in human cells.


Subject(s)
Chromosomes, Human, Pair 19 , RNA-Binding Proteins/genetics , Amino Acid Sequence , Animals , Base Sequence , Chromosome Mapping , Cloning, Molecular , DNA, Complementary , HeLa Cells , Humans , Mice , Molecular Sequence Data , RNA, Messenger , RNA-Binding Proteins/metabolism , Sequence Homology, Amino Acid , Tumor Cells, Cultured
8.
Hinyokika Kiyo ; 40(2): 131-4, 1994 Feb.
Article in Japanese | MEDLINE | ID: mdl-8128923

ABSTRACT

A rare case of benign retroperitoneal schwannoma in a 47-year-old female is reported. She was initially diagnosed to have an adrenal tumor at the Prefectural Cancer Center Hospital and was subsequently admitted to this hospital. When she was admitted, she was diagnosed as undergoing a hypercalcemic crisis (s-Ca 9.2mEq/l). Serum intact-PTH was elevated, but urine cyclic-AMP and % TRP were almost normal. Gd-DTPA enhanced T1 weighted MRI showed a hyperintensity area between the trachea and the thyroid. A parathyroid tumor and a retroperitoneal tumor were excised completely. The final pathological diagnoses were determined to be a parathyroidal adenoma and a retroperitoneal Schwannoma. After a 6-month follow-up period no recurrence and no electrolytic imbalance were detected and the patient presently displays no further symptoms.


Subject(s)
Hyperparathyroidism/complications , Neurilemmoma/complications , Retroperitoneal Neoplasms/complications , Adenoma/complications , Adrenal Gland Neoplasms/diagnosis , Diagnosis, Differential , Female , Humans , Middle Aged , Neoplasms, Multiple Primary/pathology , Neurilemmoma/diagnosis , Parathyroid Neoplasms/complications , Retroperitoneal Neoplasms/diagnosis
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