Serine protease Omi/HtrA2 targets WARTS kinase to control cell proliferation.

The serine protease Omi/HtrA2 was initially regarded as a proapoptotic molecule that proteolyses several proteins to induce cell death. Recent studies, however, indicate that loss of Omi protease activity increases susceptibility to stress-induced cell death. These complicated findings suggest that the protease activity of Omi is involved not only in apoptosis but also in cellular homeostasis. However, the targets which Omi uses to mediate this novel process are unknown. Previously, we showed that WARTS (WTS)/large tumor-suppressor 1 mitotic kinase interacts with the protein/discs-large protein/zonula (PDZ) domain of Omi and promotes its protease activity. We now report that WTS is a substrate for Omi protease activity, thus it is not only a regulator but also a downstream target of this protease. Interaction with Omi PDZ domain is required for WTS to be proteolysed. When caspase-9-deficient mouse embryonic fibroblasts (MEFs) were treated with staurosporine, WTS was proteolysed by activated endogenous Omi without induction of cell death. Therefore, protease activity of Omi and proteolysis of WTS are not necessarily required for cell death. We found that depletion of Omi from HeLa cells results in accelerated cell proliferation despite no significant change in the duration of mitosis. The depletion of WTS showed the same effect on S phase progression. Therefore, WTS proteolytic fragment(s) generated by Omi may act as an inhibitor of G1/S progression. Our data reveal a role for Omi-mediated processing of WTS in negative regulation of cell cycle progression at interphase, suggesting a novel function of Omi other than apoptosis.

[A case of post-tuberculosis chronic empyema with effective low-dose, long-term clarithromycin administration].

A 78-year-old woman with post-tuberculosis chronic empyema who underwent unsuccessful decortication in July 1993, was admitted to our hospital in November 1994 because of M. tuberculosis in her sputum. One month after anti-tuberculosis treatment, the M. tuberculosis disappeared from the sputum, but a pleurocutaneous fistula that exuded abundant bloody pus containing Pseudomonas aeruginosa persisted in the area of chronic empyema. In July 1997, 200 mg of clarithromycin (CAM) was administered daily; 21 months later, the pus and pleurocutaneous fistula had disappeared. Post-tuberculosis chronic empyema with a fistula is generally an intractable condition that necessitates surgery. In this case, the empyema did not improve during the 4 years after the surgery. However, low-dose, long-term CAM administration brought about a disappearance of pus from the fistula and closure of the empyema spaces.

[Clinical efficacy of imipenem/cilastatin sodium for respiratory infections in patients with lung cancer].

Imipenem/cilastatin sodium (IPM/CS) was administered to 102 patients with respiratory tract infections and lung cancer. Patients with other serious diseases were excluded and a total of 73 patients were enrolled. They were divided into 12 patients who underwent surgery (operated group) and 61 who did not (non-operated group); the latter group included 28 patients treated with anticancer agents or radiation therapy (treated group) and 33 untreated patients (untreated group). IPM/CS was effective in 75% of the patients, both with and without surgery. The drug was effective in 81% of the treated group, although many of the patients had Stage III or more advanced cancer, as well as bronchial occlusion. IPM/CS was also effective in 69% of the untreated group, although many of the patients have serious infections and a PS (Performance Status) of 3 or greater. Thus, IPM/CS treatment achieved good results. Bacteriological studies showed that 3 out of 4 strains in the operated group and 16 out of 18 in the non-operated group were eliminated. Safety was evaluated in all patients. Two patients (2%) experienced side effects and two others (2%) showed abnormal clinical findings, but the symptoms were mild and resolved after discontinuation or completion of therapy. In conclusion, IPM/CS was very effective for treating respiratory infections in patients with lung cancer.

[Investigation of the actual conditions of hospital nurses working on three rotating shifts: questionnaire results of shift work schedules, feelings of sleep and fatigue, and depression].

These studies were performed to clarify (1) the actual conditions concerning rotating shift schedules of nurses in Japanese university and college hospitals and to evaluate (2) some aspects of the physical and mental health, and (3) sleep profile of hospital nurses working on counter-clockwise shift rotation. Two questionnaire surveys and the OSA sleep inventory (OSA) were carried out. The subjects in the study (1) were a total of 80 nursing directors in university and college hospitals. The questionnaire covered 4 categories, such as the schedule most frequently adopted and reasons for using the schedule. The questionnaires were returned by 67 directors (83.8%). The subjects in the study (2) were 189 nurses working on three-shift work schedules at Asahikawa Medical College Hospital. The items in the questionnaire covered 7 categories, as follows: 1) feeling of sleep after each shift (8 items); 2) feeling of fatigue after each shift (30 items); 3) physical symptoms; 4) inter-personal problems; 5) all the items on Zung's self-rating depression scale (SDS); 6) all the items on the Horne and Ostberg morningness-eveningness questionnaire; and 7) 24 items on the Maudsley personality inventory. The questionnaires were returned by 156 nurses (82.5%), whose mean age and duration of shift-work employment were 27.2 +/- 5.1 and 5.0 +/- 4.3 years (mean +/- SD), respectively. For 152 nurses (97.4%) of those returning the questionnaire, the working schedule consisted of 2 consecutive night shifts and 2 consecutive evening shifts, following a variable number of day shifts (rapid and counterclockwise shift rotation). The subjects in the study (3) were 8 healthy nurses working on above-mentioned three rotating shifts at the psychiatric ward of Asahikawa Medical College Hospital, whose mean age was 29.4 +/- 5.8 years (mean +/- SD). All the subjects recorded their sleep-logs and underwent OSA everyday for 30 consecutive days. Of the 240 OSA data, 95 data (16 after day shift, 17 after the 1st night shift, 16 after the 2nd night shift, 15 after the 1st evening shift, 16 after the 2nd evening shift, 15 after day off) were analyzed. In addition to five of sleep factors in the OSA analysis, we evaluated the global score (GS), which represents subjective global feeling of sleep. In the study (1), 47 of 66 hospitals (71.2%) adopted rapid and counterclockwise shift rotation. The results of study (2) were as follows: 1) After the first night shift (diurnal sleep), the sleep problems were worst, and the frequency of taking sleep-inducing drugs was highest (12.6%); 2) Feelings of fatigue were the highest level after each of the two night shifts; 3) SDS score was relatively high (57.8 +/- 8.1, mean +/- SD); 4) The older the nurse, the greater the aggravation of both sleep problems and fatigue; 5) Sleep problems after day shifts were worse and SDS score was higher in nurses classified as "nightowls" compared to those in nurses classified as "morning people"; and 6) There was no difference between introverts and extroverts in sleep problems, fatigue or SDS score. The results of study (3) were as follows: 1) In 3 of sleep factors (Sleepiness, Integrated sleep, sleep initiation) and GS, there were significant differences among each rotating shift schedule including day off; 2) The highest score in five of sleep factors and the highest GS were noted after the 2nd night shift (nocturnal sleep), and the lowest after the 1st night shift (diurnal sleep); and 3) The younger subjects (n = 4; mean age, 24.5 years) showed a higher GS compared with that of the older subjects (n = 4; mean age, 34.2 years).

Polymerase chain reaction fiber analysis and somatic mosaicism in autopsied tissue from a man with Duchenne muscular dystrophy.

Single muscle fibers, obtained at autopsy from a 22-year-old man with Duchenne muscular dystrophy were examined immunocytochemically and also using polymerase chain reaction (PCR). Dystrophin-positive cells were widespread in skeletal, cardiac, smooth muscle, and in brain cells. PCR and Southern blot analyses of DNA from peripheral blood lymphocytes revealed a deletion of exon 45 in the dystrophin gene. With PCR of single fibers, three bands corresponding to exons 44, 45, and 47 were present in the normal control muscle fibers and dystrophin-positive fibers from the patient, while only two bands, exons 44 and 47, were observed in dystrophin-negative fibers. Therefore, in this patient, the genotype of dystrophin-positive fibers differed from that of the dystrophin-negative fibers, possibly because of a somatic mosaicism for deletion in the dystrophin gene. A mutation of the dystrophin gene may have occurred in one cell at an early stage of ontogenesis.

Characterization of antineutrophil antibodies in patients with neutropenia associated with nutritional copper deficiency.

Six antineutrophil antibody (ANA)-positive patients with copper deficiency were classified into two groups; those with (group A, n = 3) and those without (group B, n = 3) neutropenia. The percent binding of ANA for normal peripheral neutrophils was similar in both groups (83.5 +/- 7.2 vs. 79.1 +/- 10.5%). The percent binding of sera to cultured promyelocytic leukemia cells (HL-60) was increased in group A (from 3.7 +/- 3.2 to 12.2 +/- 2.3%) but not in group B (from 65.3 +/- 21.7 to 40.7 +/- 6.3%) after stimulation of HL-60 with DMSO. The stimulated HL-60 cells expressed CD 16 and CD 11b antigens. In the presence of monoclonal antibody for CD 16, the titer of ANA was nil in group A and unchanged in group B. Thus, ANA of patients with neutropenia may recognize mainly the CD 16 antigen, the Fc gamma receptor III of neutrophils.

A family of selective immunodeficiency with normal immunoglobulins: possible autosomal dominant inheritance.

We report here our findings in two Japanese siblings who experienced recurrent bacterial and viral infections since early infancy. Recent symptoms included diarrhoea, conjunctivitis, rashes, headache, sore throat, joint pain, vomiting and vertigo, all similar to those seen in toxic shock syndrome, except for shock. These symptoms improved following gammaglobulin treatment. Staphylococcus aureus with coagulase type IV was continuously isolated from nasal smears producing toxic shock syndrome toxin-1 (TSST-1). Serum antibodies did not or only poorly responded to TSST-1, diphtheria toxoid, varicella virus and rubella virus, whereas total and subclass levels of serum immunoglobulin and in vitro DNA synthesis of lymphocytes stimulated by TSST-1, Staph. aureus, varicella vaccine and mitogens were normal. In the family, ten other members in three generations (five males: five females) including the mother had similar clinical symptoms. Thus, the disease may be inherited in an autosomal dominant fashion.

Dystrophin and dystrophin-related protein in the brains of normal and mdx mice.

To clarify the localization and characterization of dystrophin and dystrophin-related protein (DRP) in the brains of normal and mdx mice, we carried out immunostaining and immunoblotting studies using four region-specific antidystrophin and anti-DRP antibodies. With immunostaining, punctate immunoreactivity of dystrophin was seen along the cell bodies and dendrites of the cerebral cortical neurons in the normal mice. By contrast, dystrophin was not detected at all in the brains of mdx mice. Immunoreactivity of DRP was observed in the vascular walls, pia mater, and choroid plexus of both normal and mdx brains, but not in the neuronal cells. The possible compensatory increase of DRP as seen in the skeletal muscles of mdx mice was not noted in the brains. The immunoblot findings were very consistent with those of immunostain. Although further studies of brain-type dystrophin are necessary, it seems unlikely that DRP participates in any physiological function of the neuronal cells.

Localisation and characterisation of dystrophin in the central nervous system of controls and patients with Duchenne muscular dystrophy.

The aim was to localise and characterise dystrophin in various human tissues, especially in the CNS. Immunoblotting and immunostaining studies were carried out with eight region-specific dystrophin antibodies. In necropsy tissue from controls, dystrophin was noted as a doublet in immunoblots of striated muscle, and as a single band in those of smooth muscle and the CNS. With immunostaining, punctate immunoreactivity was seen on the cell bodies and dendrites of the cerebral cortical neurons and cerebellar Purkinje cells. By contrast, dystrophin was not detected in any tissues, including the cerebrum and cerebellum, of patients with Duchenne muscular dystrophy who had an intellectual disturbance.

Dystrophin and dystrophin-related protein in the central nervous system of normal controls and Duchenne muscular dystrophy.

To clarify the localization and characterization of dystrophin and dystrophin-related protein (DRP) in the human central nervous system (CNS), we carried out immunoblotting and immunostaining studies using three region-specific anti-dystrophin and one anti-DRP antibodies. With immunostaining, punctuate immuno-reactivity of dystrophin was seen along the cell bodies and dendrites of the cerebral cortical neurons and cerebellar Purkinje cells in the normal controls autopsied. By contrast, dystrophin was not detected at all in the CNS of Duchenne muscular dystrophy (DMD) patients with intellectual disturbance. Immunoreactivity of DRP was observed in the vascular walls of both normal and DMD brains, but not in the neuronal cells. Compensatory increase of DRP was not noted in DMD brains. This study suggests that in DMD the brain-type dystrophin originally present in neurons is absent and may be related to the intellectual disturbance.

[A study on primarily treated tuberculosis cases with positive sputum].

A total of 377 cases with primarily treated bacillary tuberculosis selected from 432 patients admitted to 5 major national sanatoria during 1987 was analysed and compared with the same sort of studies done in national sanatoria in 1976 and 1980, and in addition 21 dead cases were investigated. The results were as follows. 1. 110 cases (29%) were over 60 years of age. 2. New intensive regimens containing INH and RFP has become popular (over about 70%). 3. The duration of chemotherapy has been shortened (65% terminated within 12 month, while only 9.5% in 1980). 4. The duration of admission has been shortened (65% discharged within 6 month, while 59% 1980). 5. 21 dead cases were in the higher age group (70% were over 60 years of age) and the higher rate of complications including diabetes mellitus (28%), cancer (21%) and heart failure (19%). 6. At the start of chemotherapy, chest X-ray showed fresh types of GAKKEN A & B in 307 cases (82%) and cavitary type of GAKKAI I & II in 274 cases (73%) and these proportions have not changed since 1980 and the response to chemotherapy was as good as in 1980. Based on the above findings, the older age patients with various complications might be regarded as the most difficult cases to be cured at present in coming years.

Gonadotropin-secreting bronchogenic carcinoma in a woman.

We report the case of a woman with a squamous carcinoma of the bronchus which secreted human chorionic gonadotrophin. Earlier reports have all involved males. Some evidence that estimation of serum HCG levels might prove a useful marker of early malignancy is presented.

Dyspnoea, disability, and distance walked: comparison of estimates of exercise performance in respiratory disease.

Forty-four patients with airway obstruction and 18 with pulmonary infiltration were studied in an attempt to correlate exercise tolerance, as assessed by a simple walking test, with basic respiratory function values and differing subjective assessments of exercise performance. The distance walked in 12 minutes was significantly correlated with the response to a structured questionnaire and with the patients' assessment of performance using an oxygen-cost diagram. The distance walked did not agree well with simple subjective estimates obtained in the clinical history. It was better correlated with forced vital capacity than with forced expiratory volume in one second in both groups of patients, and was well correlated with carbon monoxide transfer factor in those with pulmonary infiltration. The scatter of results, however, was such that exercise performance could not usefully be predicted from the respiratory function values or from subjective assessments. Simple exercise tests are an essential part of assessing disability and response to treatment in patients with respiratory impairment.