A case of primary squamous cell carcinoma of the endometrium associated with extensive "ichthyosis uteri".

BACKGROUND: Ichthyosis uteri is an uncommon entity in which the entire endometrium is replaced by stratified squamous epithelium. Though the condition often is considered as benign, dysplastic changes have been reported. CASE: We describe herein an exceedingly rare case of primary squamous cell carcinoma of the endometrium (PSCCE) associated with extensive ichthyosis uteri with chronic pyometra, who presented with blood-stained vaginal discharge of six-seven months duration. Although repeated endometrial biopsies revealed only strips of stratified squamous epithelium showing moderate to severe dysplastic changes, the tumor markers and magnetic resonance imaging strongly suggested advanced uterine body malignancy. Exploratory laparotomy was performed, and histologic findings of the superficial layer were consistent with ichthyosis uteri; in contrast the lesion of invasive squamous cell carcinoma was located in the deeper layer and lymph nodes. No dysplastic changes of the cervix were noted. CONCLUSIONS: It is suggested that PSCCE could be associated with pre-existing ichthyosis uteri and deeper biopsies should be performed for the accurate preoperative diagnosis of cases with chronic pyometra.

[Two cases of exercise-induced acute renal failure with idiopathic renal hypouricemia].

Acute renal failure without oliguria developed in a 25-year-old male and a 19-year-old male after exercise. Marked hypouricemia became apparent during improvement of their renal function. Increased excretion of uric acid into the urine, increased fractional excretion of uric acid(clearance ratio of uric acid against creatinine), and normal concentration of plasma xanthine and hypoxanthine were observed in both cases. Probenecid and pyrazinamide loading test suggesting decreased reabsorption of uric acid in the proximal convoluted tubules revealed that presecretory reabsorption defect of uric acid resulted in the hypouricemia in both cases. These two cases were diagnosed as having idiopathic renal hypouricemia.

Ultraviolet B (UVB) light-induced histamine release from rat peritoneal mast cells and its augmentation by certain phenothiazine compounds.

When rat peritoneal mast cells were exposed to ultraviolet (UV) light (UVA, UVB and UVC), histamine release was evoked in a dose (intensity X time) dependent manner. The potency order of UV light in inducing the histamine release was UVC > UVB >> UVA. In this study, we focused on the effect of ultraviolet B (UVB) on histamine release from rat mast cells. The UVB-induced histamine release occurred at doses higher than 7.8 kJ m(-2), even at 4 degrees C. At a UVB dose of 18.8 kJ m(-2), where a 51.9+/-4.8% histamine release and a 58.8+/-6.8% degranulation took place, Trypan blue-stained cells accounted for 14.4+/-1.3% of the cells, and the lactate dehydrogenase (LDH) release was about 4.9+/-2.8%. This suggests that the membrane permeability to low molecular weight substances was increased by UVB exposure. The UVB-induced histamine release was inhibited by ascorbic acid at concentrations higher than 500 microM, suggesting the involvement of a radical reaction in the process. The UVB-induced histamine release was enhanced by some phenothiazine compounds, i.e., promethazine, trimeprazine, mequitazine, chlorpromazine, trifluoperazine, ethopropazine and thioridazine. We conclude that the phototoxicity of phenothiazine compounds may be due in part to an enhancement of UVB-induced histamine release from mast cells.

High-level production of recombinant human parathyroid hormone 1-34.

Expression of the synthetic human parathyroid hormone 1-34 [hPTH(1-34)] gene by a gene fusion strategy was demonstrated. hPTH(1-34) was produced at the C terminus of the partner peptides involving amino acids 1 to 97, 1 to 117, or 1 to 139 of a modified Escherichia coli beta-galactosidase by linker peptides containing oligohistidine of different lengths. The fusion proteins in the inclusion bodies were rendered soluble with urea and subjected to site-specific cleavage with the secretory type yeast Kex2 protease. Optimal expression and enzymatic processing were achieved in the fusion protein beta G-117S4HPT, constructed from amino acids 1 to 117 of beta-galactosidase and the linker of HHHHPGGSVKKR. The fusion protein accumulated more than 20% of the E. coli total protein. The hPTH(1-34) was purified up to 99.5% with a good yield of 0.5 g/liter of culture. The purified product was identified as intact hPTH(1-34) by amino acid analysis and N-terminal sequencing.

Hyperproduction of a recombinant fusion protein of Staphylococcus aureus V8 protease in Escherichia coli and its processing by OmpT protease to release an active V8 protease derivative.

The expression of a recombinant fusion protein including Staphylococcus aureus V8 protease was studied by using Escherichia coli as the host strain. When the mature V8 protease was expressed as a fusion protein with a truncated E. coli beta-galactosidase (beta-gal97S4D), we could not obtain a sufficient amount of the enzyme because of the toxicity resulting from the expressed protease activity. Synthesis of V8 protease was increased by constructing a sandwich-type fusion protein consisting of beta-gal97S4D, a V8 protease derivative with the 56 C-terminal amino acids deleted (V8 delta 56) and a truncated aminoglycoside-3'-phosphotransferase. This fusion protein was successfully produced as inactive inclusion bodies. To release the V8 delta 56 protease from the fusion protein, we developed a novel processing method using an endogeneous E. coli OmpT protease, which can recognize the dibasic amino acid residues located in the linker peptides of the fusion protein. After solubilizing the inclusion bodies with urea, the V8 delta 56 protein was automatically released from the fusion protein by the OmpT protease, which was coprecipitated with the inclusion bodies. The V8 delta 56 protease thus obtained showed the same enzymatic activity as that of the native V8 protease. We demonstrate in this study that the N-terminal prepro sequence and the C-terminal repeated sequence of this enzyme are not necessary for its enzymatic activity and protein folding.

Mutational study at Ser300 position of the Escherichia coli lactose repressor.

We have previously reported that a Ser300Asn mutant of the Escherichia coli lactose repressor protein produced a temperature-sensitive phenotype. In order to analyze the structure-function relationship of the lactose repressor protein, we conducted 18 amino acid substitutions at this Ser 300 site by using in vitro mutagenesis. The substitutions at this position that exhibited repressors with the wild-type phenotype in vivo were Gly, Ala, Ile, Thr, Met and Cys; the other 10 substitutions examined (Leu, Val, Tyr, Trp, Asp, Glu, Gln, His, Lys and Arg) resulted in the lacI- phenotype. In addition, the Ser300Phe mutation resulted in the lacIts phenotype, while the Ser300Pro mutation resulted in lacIts,s. It seems likely that the Ser300 position plays an important role in oligomer formation and inducer binding.

Thermo-inducible expression of a recombinant fusion protein by Escherichia coli lac repressor mutants.

The lac I gene of Escherichia coli encodes the lactose repressor. We isolated temperature-sensitive mutants of the lac I gene by in vitro mutagenesis with hydroxylamine. The new mutation sites were determined, and replacement of a single amino acid had respectively occurred at amino acid positions 241 (Ala-->Thr), 265 (Gly-->Asp) and 300 (Ser-->Asn). These mutation sites were located in the core region of the lac repressor protein. Temperature-dependent expression of beta-galactosidase was observed in the strains having these mutant lac I genes. By using these temperature-sensitive lac I genes, we developed a thermo-inducible expression system for a foreign gene under the control of the lac promoter. A recombinant fusion protein, consisting of a derivative of E. coli beta-galactosidase and the human calcitonin precursor peptide, was efficiently produced by using this system.

High expression of a recombinant human calcitonin precursor peptide in Escherichia coli.

Human calcitonin (hCT) is a C-terminus alpha-amidated peptide hormone consisting of 32 amino acids. The amidated structure is essential for its biological activities, and the C-terminal-glycine-extended precursor peptide, hCT[G], is converted to bioactive hCT by a C-terminus-alpha-amidating enzyme. An efficient production method is described for the hCT[G] peptide, as a part of the fusion protein consisting of a modified E. coli beta-galactosidase, linker amino acids and hCT[G]. Stable inclusion bodies of the fusion protein in E. coli were expressed by focusing on the amino acid charge, and the fusion protein was modified by inserting a basic amino acid sequence into its linker region. This modification greatly affected the formation of inclusion bodies. E. coli strain W3110/pG97S4DhCT [G]R4 could produce a large amount of stable inclusion bodies, and the hCT[G] peptide was released quantitatively from the fusion protein by S. aureus V8 protease. This enabled a large-scale production method to be established for the hCT[G] precursor peptide in E. coli to produce mature hCT.

Cloning and expression of the gene encoding the glutamic acid-specific protease of Streptomyces griseus ATCC10137.

The gene (sgpE) encoding the Streptomyces griseus glutamic-acid-specific protease (SGPE) was cloned and sequenced. The sgpE gene contained an open reading frame of 1065 nucleotides encoding 355 amino acids (aa) with a pre-propeptide of 168 aa ending at Glu, suggesting the probability of auto-proteolysis. A Streptomyces lividans strain carrying the plasmid-borne sgpE under control of the traA gene promoter secreted mature SGPE into the culture medium. Compared to Staphylococcus aureus protease V8, the purified SGPE was more resistant to urea. It is suggested that SGPE would be a useful tool for site-specific processing of proteins, even under denaturing conditions.

[Evaluation of cardiotoxicity by radionuclide angiocardiography in the chemotherapy of gynecologic malignancies].

Cardiotoxicity was evaluated by multigated radionuclide angiocardiography using in vivo 99mTc labeling of red blood cells in the multiagent chemotherapies of gynecologic malignancies. The left ventricular ejection fraction (LVEF) was determined by computer-assisted analysis of left ventricle time activity curves (TAC) of the angiocardiograms. The mean age of fifteen patients was 52.3 (38-66) years. Ovarian carcinoma (13 patients), the endometrial carcinoma (one) and uterine leiomyosarcoma (one) were treated with a total of 69 combination chemotherapy of CAP every 4 weeks. Fifteen patients underwent a total of 76 quantitative radionuclide angiocardiograms (three to seven studies per patient). The values of LVEF before treatment were 57.5 +/- 8.3 (mean +/- SD) (48.7-75.4), which were greater than those of normal persons (45). The values of LVEF during treatment were 44.9 +/- 6.2 (mean +/- SD) (37.6-58.5), which were significantly (p less than 0.001) lower than the pretreatment values. The decrease rates of LVEF in fifteen patients were 21.0 +/- 10.4% (mean +/- SD). Over 20% decrease of LVEF was observed in eight patients (53.3%) following chemotherapies. The patients showing a decrease of the values of LVEF were treated with lower dosage of chemotherapeutic agents in three, changed in seven and discontinued in six the agents. Nine patients who changed the chemotherapeutic regimens showed improvement of the values of LVEF in six (66.7%), no change in two (22.2%) and deterioration in one (11.1%). The values of LVEF were decreased in four patients (44.4%) at 6.7 (3-11) months following chemotherapies. It is suggested from these findings that the determination of left ventricular ejection fraction using multigated radionuclide angiocardiography may prevent development of cardiotoxicity before, during and after the chemotherapies of gynecologic malignancies and allow a clinically important assessment of the cardiotoxicity of the chemotherapeutic agents.

[The effect of nonanoyl vanillylamide on the chemically-induced nociception].

The aim of the present investigation was to determine the effects of a single dose and serial doses of nonanoyl vanillylamide (NVA) administered subcutaneously on responses induced by chemical stimuli in the mouse. The treatment with a single subcutaneous injection produced a significant decrease in the number of abdominal contraction responses produced by intraperitoneal injections of phenylbenzoquinone and in the responsive time of licking induced by plantar injection of formalin. The serial subcutaneous administration of NVA caused a dose related antinociceptive effect against a chemical nociceptive stimulus using the phenylbenzoquinone-induced writhing test, while NVA was ineffective against a chemical stimulus, formalin. These results suggest that serial doses of NVA administered peripherally cause different effects in two types of chemically-induced nociception.