Comparison of clinical characteristics of patients with Mycobacterium avium complex disease by gender.

The clinical characteristics of male patients with pulmonary Mycobacterium avium complex disease have not been clearly defined. We aimed to clarify the clinical characteristics of male patients with pulmonary Mycobacterium avium complex disease compared with female patients.We retrospectively reviewed the medical records of patients with pulmonary Mycobacterium avium complex disease who visited the outpatient clinic of the Shinshu University Hospital between 2003 and 2016 and compared the clinical characteristics of male and female patients.A total of 234 patients with pulmonary Mycobacterium avium complex disease were identified (68 men and 166 women). Male patients were significantly older than female patients. Blood examination results showed that the lymphocyte count, total protein level and albumin level were significantly lower in men than in women. Chest imaging findings were broadly categorised into the fibrocavitary and nodular bronchiectasis types. There were no significant differences in chest imaging findings and the time from diagnosis to disease exacerbation between men and women.During the study period, the incidence of the nodular bronchiectasis type of pulmonary Mycobacterium avium complex disease in male patients increased compared with previous reports. Men had no difference in time to exacerbation compared with women.

Effects of petrissage massage on fatigue and exercise performance following intensive cycle pedalling.

OBJECTIVE: Petrissage is assumed to influence circulation as well as interstitial drainage of both superficial and deep tissues. To study its effect it was applied between consecutive bouts of supramaximal exercise performed by the lower leg muscles. METHODS: Subjects were 11 healthy female students actively engaged in sports. Exercise bouts of ergometer cycling at loads determined individually (0.075 kp x body weight (kg)) for 5 s repeated eight times at intervals of 20 s had to be performed twice on an experimental day with 35 min intermittent bed rest. Each subject was investigated on two occasions with a minimum interval of 1 week, once without (control, CO) and once with 10 min petrissage (massage, MA) of the exercising lower leg during the bed rest phase. Effects of exercise bouts on blood lactate, muscle stiffness and perceived lower limb fatigue and their recovery before and after the second exercise bout were determined. RESULTS: For the first exercise bouts total power did not differ between MA and CO. Courses of blood lactate did not differ between MA and CO. However, recovery from measured muscle stiffness (p<0.05) and perceived lower limb fatigue (p<0.05) were more pronounced and total power during the second exercise bout was enhanced (p<0.01) in MA as compared with CO subjects. CONCLUSION: Petrissage improved cycle ergometer pedalling performance independent of blood lactate but in correlation with improved recovery from muscle stiffness and perceived lower limb fatigue.

Induction of photoreceptor-specific phenotypes in adult mammalian iris tissue.

We show that iris tissue in the adult rat eye, which is embryonically related to the neural retina, can generate cells expressing differentiated neuronal antigens. In addition, the Crx gene transfer induced the specific antigens for rod photoreceptors in the iris-derived cells, which was not seen in the adult hippocampus-derived neural stem cells. Our findings demonstrate a remarkable plasticity of adult iris tissue with potential clinical applications, as autologous iris tissue can be feasibly obtained with peripheral iridectomy.

Health-related quality of life in patients with pulmonary non-tuberculous mycobacteria infection.

BACKGROUND: The health-related quality of life (HRQoL) in patients with pulmonary non-tuberculous mycobacteria (pNTM) infection has not yet been quantified. OBJECTIVE: To evaluate the reliability and validity of the COPD Assessment Test (CAT) and St George's Respiratory Questionnaire (SGRQ) in quantifying the HRQoL of patients with pNTM. METHODS: We performed a cross-sectional study of 52 patients with pNTM. All the subjects completed the CAT, SGRQ and Short Form-36 Health Survey (SF-36) questionnaires and underwent pulmonary function testing (PFT). A test-retest was performed and Cronbach α was calculated to assess reliability. Correlations of the CAT and SGRQ with SF-36 and PFT were performed to assess validity. RESULTS: The mean age of the patients was 67 years; 96% (50/52) were female. Both individual and total CAT and SGRQ scores showed good correlation between the test on Day 1 and the repeat test on Day 5. Cronbach's α (0.77-0.92) indicated satisfactory internal consistency. All scores were moderately or strongly correlated with the SF-36 Physical Component Summary score. CONCLUSION: The SGRQ and CAT questionnaires showed statistically significant validity in assessing HRQoL in patients with pNTM.

c-myc and p53 gene expression in the differentiation of temperature-sensitive mutants of teratocarcinoma F9 cells.

We have previously reported the isolation of several temperature-sensitive (ts) mutants of F9 cells. Further investigations showed that some mutants were induced to differentiate at non-permissive temperature of cell growth, accompanied by changes in the expression of various genes, whereas others were not. During the differentiation induced by shifting up to the non-permissive temperature, a rapid and transient decrease in both c-myc and p53 mRNA levels and rapid induction of c-jun mRNA were observed. These changes were specific in differentiation-inducible mutants and were not observed in a non-inducible mutant. In both types of mutants, the level of c-myc mRNA decreased in association with growth retardation at the non-permissive temperature. The p53 mRNA, however, showed specific increase in the differentiation-inducible ts mutants. These observations suggest distinct roles for p53 and c-myc from proliferation to differentiation in teratocarcinoma stem cells.

Nucleotide sequence encoding human pancreatic ribonuclease.

A cDNA coding for human pancreatic ribonuclease was isolated from a pancreas cDNA library and sequenced. This cDNA (1620 bp) includes an entire open reading frame encoding mature protein (128 aa) following a signal peptide (28 aa) as well as 5'- and 3'-untranslated regions.

Molecular cloning and expression of human ribonuclease 4 cDNA.

A cDNA coding for human ribonuclease 4 was isolated from a pancreas cDNA library and sequenced. This cDNA (996 bp) includes an entire open reading frame encoding mature protein (119 aa) following signal peptide (28 aa). Expression of mature protein in Escherichia coli showed an apparent molecular mass of about 16 kDa, which was slightly lower than the mature form of human RNase 1, in SDS-PAGE.

The humanized anti-HM1.24 antibody effectively kills multiple myeloma cells by human effector cell-mediated cytotoxicity.

A mouse monoclonal antibody, anti-HM1.24 (IgG2a/kappa), binds to a surface antigen preferentially overexpressed on multiple myeloma (MM) cells, and exhibits potent antitumor cell activity against MM cells by antibody-dependent cell-mediated cytotoxicity (ADCC). To develop an antibody-based immunotherapy against MM, a humanized anti-HM1.24 antibody, in which all FRs correspond to naturally processed human FRs, has been successfully constructed with the aid of both the hybrid variable region and two-step design methods. This humanized anti-HM1.24 antibody (IgG1/kappa) is able to effectively induce ADCC against human myeloma KPMM2 and ARH77 cells in the presence of human PBMCs as effectively as a chimeric anti-HM1.24 antibody. The humanized anti-HM1.24 antibody, therefore, could be expected as a potent immunotherapeutic agent for MM patients.

Effects of erythroid differentiation factor (EDF) on proliferation and differentiation of human hematopoietic progenitors.

The effects of erythroid differentiation factor (EDF) on normal human hematopoietic progenitor cells were examined by bone marrow colony assay. Addition of EDF to the erythroid colony assay system enhanced erythroid burst-forming unit (BFU-E)-derived colony formation, and this effect disappeared on removal of adherent cells. Conditioned medium of EDF-treated monocytes also enhanced BFU-E colony formation, whereas conditioned medium of EDF-treated T cells did not. In contrast, EDF inhibited erythroid colony-forming unit (CFU-E) colony formation dose-dependently, although it had no effect on colony formation by myeloid cells. These data show that EDF has a specific effect on human hematopoietic progenitors of the erythroid lineage. The results also indicate that EDF enhanced BFU-E colony formation by stimulating adherent cells to produce factors with burst-promoting activity (BPA), but suppressed CFU-E colony formation by promoting differentiation of these cells.

Effects of activin A on deoxyribonucleic acid synthesis, iodine metabolism, and cyclic adenosine monophosphate accumulation in porcine thyroid cells.

We have recently shown the presence of activin A in human thyroid cells. To determine whether activin A affects the growth and function of the thyroid, we investigated its effects on DNA synthesis, iodine metabolism, and cAMP accumulation in cultured porcine thyroid cells. Activin A increased DNA synthesis. This effect was abolished by addition of follistatin but additively enhanced by epidermal growth factor. In contrast, activin A significantly reduced uptake and release of iodide by the thyroid and also TSH-induced cAMP accumulation but did not inhibit the cAMP accumulation induced by forskolin. These data indicate that activin A stimulates thyroid growth irrespective of cAMP accumulation and inhibits thyroid function.

A sensitive enzyme immunoassay for anti-thyroglobulin antibody using Fab'-horseradish peroxidase conjugate. Evaluation of in vitro anti-thyroglobulin antibody synthesis by lymphocytes from patients with autoimmune thyroid disease.

A sensitive enzyme immunoassay (EIA) for the detection of anti-thyroglobulin (Tg) antibodies was developed using Fab'-horseradish peroxidase (HRP) conjugate. Anti-Tg antibody was assayed by incubation with a thyroglobulin-coated polystyrene ball and then with affinity-purified anti-IgG Fab'-HRP conjugate. The HRP activity was assayed fluorimetrically. The sensitivity was 625 amol/tube and anti-Tg antibody levels between 0.5 and 200 ng/ml could be determined. The recoveries of anti-Tg antibody added to human sera at three different concentrations were 94.2-101.0%. Both within- and between-assay coefficients of variation were below 10%. Significant correlation was observed between values by the EIA and TGHA method (Kendall's rank correlation coefficient = 0.712, P less than 0.001). The present EIA for anti-Tg antibody is sensitive enough to determine anti-Tg antibody synthesized in vitro by the lymphocytes from patients with autoimmune thyroid disease and normal subjects. The amounts of anti-Tg antibody synthesized by peripheral lymphocytes from patients with Hashimoto's disease were significantly greater than those from patients with Graves' disease, although serum levels of anti-Tg antibody were usually elevated in both groups of patients. The results obtained suggest that anti-Tg antibody is synthesized in a different manner in patients with Hashimoto's disease and in patients with Graves' disease.

Induction of teratocarcinoma F9 cell differentiation with cis-diammine dichloroplatinum(II) (CDDP).

cis-Diammine dichloroplatinum(II) (CDDP) is the salt of a platinum compound which has been noted to have a wide spectrum of activity against malignant disorders. We have studied the effects of CDDP on embryonal carcinoma F9 cell differentiation. In the presence of this agent in vitro, the cells showed rapid morphological changes, a marked increase in the mRNA expression of various differentiation markers accompanied by a loss of tumorigenicity. These results indicate that the differentiation of F9 cells is induced with CDDP.

Tetramer Bence Jones protein in the immunoproliferative diseases. Angioimmunoblastic lymphadenopathy, primary amyloidosis, and multiple myeloma.

The authors report the clinical features and the results of immunochemical studies of patients with angioimmunoblastic lymphadenopathy, primary amyloidosis, and multiple myeloma, each of whom had in the serum and urine multiple forms of Bence Jones protein (BJP). The BJPs were isolated and purified and were shown by electrophoretic, gel filtration, and ultracentrifugal analyses to exist as tetramers and dimers. The components in two cases were kappa type and in one lambda type. The kappa tetramers consisted of two covalent dimers and the lambda tetramer two noncovalently bound dimers present in both serum and urine.

An S-alkylating reagent with positive charges as an efficient solubilizer of denatured disulfide-containing proteins.

A novel S-alkylating reagent, N-(3-bromopropyl)-N,N,N',N',N'-pentamethyl-1,3-propanedi(ammonium bromide) (TAP2-Br) which carries two positive charges in the molecule, was prepared to increase the solubility or to decrease the hydrophobicity of cysteine-containing denatured proteins (or peptides). S-Alkylation with TAP2-Br introduces two positive charges per cysteine residue, which will effectively shift the net charge of a protein in the positive direction. Disulfide-containing proteins, such as hen egg-white lysozyme, RNase A, BSA, and soybean trypsin inhibitor (Kunitz type), were reduced and S-alkylated with TAP2-Br to evaluate the potential of this reagent compared with other S-alkylating reagents such as monoiodoacetic acid, bromosuccinic acid and (3-bromopropyl)trimethylammonium bromide. The solubilities of these denatured proteins in the pH range of 2-10 indicated that S-alkylation with TAP2-Br effectively solubilized not only basic proteins (lysozyme and RNase) but also an acidic protein containing a fairly large number of cysteine residues (BSA). Moreover, the retentions of cysteine-containing tryptic peptides derived from lysozyme on reversed-phase HPLC were greatly reduced by S-alkylation with TAP2-Br. These results indicate that TAP2-Br is very useful to increase the solubility of some cysteine-containing denatured proteins and to decrease the hydrophobicity of peptides containing cysteine residue(s).

Localization of FGFR-1 in axotomized and peripheral nerve transplanted ferret retina.

Retinal ganglion cells (RGCs) survive axotomy and regenerate their axons into the peripheral nerve graft in adult mammals. To understand the potential role of fibroblast growth factors (FGFs) in survival and regeneration of axotomized RGCs, we examined FGF receptor 1 (FGFR-1) localization in normal and PN grafted ferret retinas by immunohistochemistry in combination with retrograde labeling. Prominent expression of FGFR-1 was observed in outer plexiform layer and ganglion cell layer of normal ferret retina. In the ganglion cell layer, FGFR-1 immunoreactivity was detected in about 30% of RGCs, predominantly in large cells. In the PN grafted ferret retina, 90% of RGCs with regenerated axons expressed FGFR-1. Our findings suggest that FGFs may play an important role in the survival and axonal regeneration of RGCs of adult mammals.

Analysis of circulating hematopoietic progenitors in patients with chronic renal failure under hemodialysis.

Circulating hematopoietic progenitors were analyzed in patients with chronic renal failure (CRF) under hemodialysis (HD) by methylcellulose culture containing interleukin-3 (IL-3) to clarify the differences in hematopoiesis between patients with and without CRF-associated anemia and between good responder whose hematocrit (Ht) was preserved in more than 25% under erythropoietin (Epo) treatment and poor responders whose Ht remained less than 25% even under Epo treatment. The numbers of peripheral blood (PB) erythroid burst-forming units (BFU-E) and granulocyte-macrophage colony-forming units (CFU-GM) in HD patients without Epo treatment, whose Ht levels were greater than 30%, were similar to those in normal subjects. However, these numbers in HD patients who required Epo treatment were significantly lower than those in normal subjects. The number of PB BFU-E in HD patients who showed a poor response to Epo was significantly lower than that in HD patients who showed a good response to Epo. The number of PB BFU-E was well correlated with the number of PB CFU-GM in all groups of HD patients. There also existed a definite correlation between these numbers and the Ht levels in HD patients without Epo treatment, but not those in HD patients with Epo treatment. The sensitivity of PB BFU-E to IL-3 was lower in HD patients who showed a poor response to Epo than in the other HD patients and normal subjects. These findings indicate that hematopoiesis in HD patients with CRF associated anemia is suppressed in both the erythroid and myeloid lineage at primitive stages, and that the lower sensitivity of PB BFU-E to IL-3 in HD patients with a poor response to Epo may be associated with this poor response. In addition, the level of the serum transferrin receptor (sTfR) in HD patients without severe anemia was higher than that in normal subjects and HD patients who required Epo treatment, indicating that erythropoiesis in HD patients who do not require Epo treatment is more active than that in normal subjects and other HD patients.

Geographic clustering of myeloma in Tokushima.

Between 1978-1988, seven cases of multiple myeloma were found in T Town (population 9,000), which is located at the mouth of a large river within the boundaries of Tokushima City, Japan. This is a significantly high incidence, 7.06 per 100,000 as compared with an incidence of 1.20 in Tokushima City itself and 1.53 in Japan as a whole. The clinical and laboratory features of these patients were typical of myeloma. Although the causes of this small but apparent cluster of multiple myeloma remain obscure, the fact that five of the seven patients had been working for long periods as fishermen was notable.

Superantigens and autoantigens may be involved in the pathogenesis of gastric mucosa-associated lymphoid tissue lymphoma.

To clarify the origin of tumor cells and the possible role of antigens in the pathogenesis of mucosa-associated lymphoid tissue lymphoma (MALTL) of the stomach, we analyzed the DNA sequences of the immunoglobulin (Ig) variable region gene from tumor cells of 4 patients with low-grade and 2 patients with high-grade MALTL associated with Helicobacter pylori infection. There were few somatic mutations in the Ig variable region gene, but intraclonal variations were observed in 2 of the 4 low-grade MALTL cases. In the remaining 2 low-grade MALTL and 1 of the 2 high-grade MALTL cases, somatic mutations and intraclonal variations were evident. In contrast, somatic mutations in the Ig variable region gene were prominent, but intraclonal variation was absent in the other high-grade MALTL cases. The deduced amino acid sequences of the antigen-binding fragments (Fab) from 2 MALTL cases revealed homology with anti-thyroglobulin antibodies, 3 MALTL cases with lupus anti-DNA antibodies, and 1 MALTL case with a rheumatoid factor. Furthermore, the heavy-chain variable region 3 (V(H)3) family genes were used in 5 of the 6 MALTL cases and had conserved amino acid residues for binding to staphylococcal protein A (SpA), a superantigen of B cells. Considering that another superantigen, protein Fv, competes for binding to Fab with SpA and has been shown to play a major role in immune defenses against gut pathogens, SpA and possibly protein Fv may contribute to the development of MALTL. Thus, these observations suggest that most gastric MALTLs arise from memory B cells that are preliminarily activated by superantigens and autoantigens.

Suppression of hematopoiesis by activated T-cells in infectious mononucleosis associated with pancytopenia.

Two patients with primary Epstein-Barr virus (EBV) infection followed by pancytopenia were studied. They showed increased numbers of DR-positive, activated T-cells and serological evidence of persistent EBV infection over a 12 and 18 week period. Bone marrow granulocyte-macrophage colony formation (CFU-GM) was investigated by limiting dilution assay (LDA) and methylcellulose assay. CFU-GM of bone marrow mononuclear cells (BMMNC) was markedly suppressed in both patients during the pancytopenic phase. Removal of bone marrow T-cells by E-rosetting resulted in a significant increase of CFU-GM to normal levels in BMMNC of both patients, while no significant increase was observed in the BMMNC of normal subjects. CFU-GM in BMMNC from Patient 1 in the recovery phase was normal when DR-positive T-cells were within normal levels, irrespective of the presence or absence of T-cells. These results suggest that pancytopenia due to infectious mononucleosis in these patients was due to bone marrow suppression by activated T-cells. In vitro studies with Con A activated T-cells and their culture medium showed that suppression of CFU-GM by T-cells was mediated by a lymphokine.