Mouse CAF1, a mouse homologue of the yeast POP2 gene, complements the yeast pop2 null mutation.

The yeast POP2 protein (Pop2p) is a component of a global transcription regulatory complex and is required for gene expression of many genes in Saccharomyces cerevisiae. We constructed POP2 deletion plasmids encoding various Pop2p regions under the native POP2 promoter and found that the minimum functional region was located in two-thirds of the carboxyl terminal region. A mouse homologue of the POP2 gene (mCAF1), which corresponds to the Pop2p minimum region, partially rescued the growth defect of pop2 null mutant cells. Addition of the Pop2p amino terminal region to mCAF1 strengthened the suppression. mCAF1 also weakly suppressed the relatively high expression of the SUC2 gene of pop2 cells under glucose-repressing conditions; however, it failed to suppress the defect of full expression of the SUC2 gene under glucose-derepressing conditions. Our findings clearly demonstrate that a mammalian homologue can substitute for the yeast POP2 gene in some aspect.

Autonomic nerves containing substance P in the aqueous outflow channels and scleral spur of the guinea pig.

PURPOSE: To study the innervation of the aqueous outflow channels and scleral spur by autonomic nerves containing substance P. METHODS: The experiments were conducted on guinea pigs. Immunohistochemical techniques and capsaicin-ablation of the sensory nerves were used to investigate nerves containing substance P at the light and electron microscopic level. RESULTS: Nerves containing substance P were observed in the aqueous outflow channels and scleral spur regions. The fine structures of these nerves had a similar pattern in those regions, and the labeled elements had abundant small vesicles, a few large vesicles, and numerous neurotubuli. Following capsaicin treatment, these nerves remained intact and no degenerated substance P-like immunoreactive nerves were found. CONCLUSIONS: Nerves containing substance P are most likely of autonomic origin in view of their ultrastructural features. These nerves innervate the aqueous outflow channels and scleral spur, and are probably important for neurogenic influences on the intraocular pressure by the autonomic nervous system.

[Cutaneous polyarteritis nodosa associated with streptococcal infection in a child and a review of the literature].

Polyarteritis nodosa (PAN) is a multisystem inflammatory disease associated with necrosis of small and medium arteries. It can occur in a systemic form with manifestations in skin, joints, heart, nervous system, gastrointestinal tracts, lungs and kidneys. It is relatively rare in adults and even more in children. It may be divided into a cutaneous form (C-PAN) and a more generalized form (G-PAN). Usually C-PAN is thought to run a benign course. But some cases were reported to run chronic courses and be diagnosed G-PAN. It has been reported that some adults with PAN are related to HBV infection. On the other hand most children with PAN had upper respiratory infections some with streptococcal infections. We described a 5-year-old boy who was suffering from high fever, joint pain, subcutaneous nodular and livedo reticularis. We diagnosed him C-PAN by examination and clinical course. Anti-streptolysin O (ASO) and anti-streptokinase (ASK) increased and we suspected that C-PAN followed after streptococcal infection. We reviewed the literatures of C-PAN and streptococcal infection in childhood. Streptococcal infection has been implicated by a positive throat swab or an increase in ASO. There were 25 cases (75.7%) which were thought to be related with prior streptococcal infection in 33 cases. We thought that C-PAN may be associated with streptcoccal infection in childhood.

Ultrastructural identification of substance P containing nerves in the guinea pig trabecular meshwork.

We aimed to provide a detailed analysis of substance P (SP)-containing nerves in the trabecular meshwork by ultrastructural immunohistochemistry and capsaicin treatment for chemical ablation of the sensory nerves. Numerous myelinated and unmyelinated nerves were observed inside the sheets and intertrabecular spaces of the trabecular meshwork, and the inner side of Schlemm's canal. SP-like immunoreactive products were identified in some of these nerves and associated with numerous vesicles of different sizes, a few mitochondria and numerous neurotubili. After the capsaicin treatment, SP-like immunoreactive nerves persisted and no degenerated SP-like immunoreactive nerves were noted. On the basis of the ultrastructural features and the results of capsaicin treatment, it seems that these nerves are most probably autonomic in origin. Autonomic efferent SP-containing nerves are confirmed directly located in the trabecular meshwork; these have not been previously described at electron microscopic level. SP in autonomic efferent nerves might act as a neurotransmitter or neuromodulator for intraocular pressure regulation.

Functional and structural features of the holin HOL protein of the Lactobacillus plantarum phage phi gle: analysis in Escherichia coli system.

Lactobacillus plantarum phage phi gle has two consecutive cell lysis genes hol-lys (Oki et al., 1996b). In the present study, functional and structural properties of the hol protein (Hol) were characterized in Escherichia coli. Electron microscopic examinations showed that hol under plac in E. coli XL1-Blue injured the inner membrane to yield empty ghost cells with the bulk of the cell wall undisturbed. Northern blot analysis indicated that hol-lys genes under plac were co-transcribed, although the amount of hol transcript was larger than that of lys, ceasing via an apparently rho-independent terminator just downstream of hol. However, deletion and/or fusion experiments suggested that: (1) the N-terminal half of phi gle Hol composed of three putative transmembrane domains may be responsible for interaction with membrane; (2) the N-terminal end (five amino acids) seems nonessential; and (3) the C-terminal half containing charged amino acids appears to be involved in proper hol function. These results suggest that phi gle Hol is a member of the lambdoid holin family, but divergent in several properties from lambda holin.

[Syndrome of inappropriate secretion of antidiuretic hormone associated with miliary tuberculosis].

An 82-year-old man with the chief complaint of anorexia was referred on suspicion of pulmonary tuberculosis. He had undergone thyroidectomy because of thyroid cancer 5 years ago, had taken levothyroxine sodium, and had kept plasma level of thyroidal hormone within normal range. He had never pointed out hyponatremia. On laboratory findings on admission, serum natrium level was 125 mEq/l. A chest X-ray film showed the infiltration in both lower lung fields, and a chest CT scan revealed a miliary pattern in both lungs. Tubercle bacilli were detected from the sputum by the Ziehl-Neelsen staining. Antituberculous drugs were started. On 5th hospital day, he developed consciousness disturbance, and the serum level of natrium and osmolarity was 103 mEq/l and 250 mOsm /kgH2O, respectively, while plasma ADH level was increased to 5.9 pg/ml, and urine level of natrium and osmolarity was 123 mEq/l and 394 mOsm/kgH2O, respectively. His mental disturbance and hyponatremia gradually improved by supplementing NaCl. We diagnosed this case as SIADH associated with miliary tuberculosis. SIADH should be considered when hyponatremia was occurred in the case of miliary tuberculosis.

[Diagnosis of pulmonary aspergillosis in cases with cavity as sequela of tuberculosis].

Non-invasive pulmonary aspergillosis is frequently seen in cases with cavity as sequela of tuberculosis. Diagnosis of non-invasive pulmonary aspergillosis is not easy except for cases whose chest X-ray findings show a typical fungus ball. How to diagnose pulmonary aspergillosis was studied in 21 patients who showed changes in the radiological findings of cavity as sequela of tuberculosis. We made sputum culture of Aspergillus, measurement of precipitin antibody titer to Aspergillus fumigatus and measurement of PASTOREX ASPERGILLUS in serum and sputum on the 21 patients (group A). PASTOREX ASPERGILLUS in sputum was evaluated in three grades: 1+ approximately 3+. We diagnosed 14 patients as pulmonary aspergillosis (PA group) among the 21 patients taking into account the chest X-ray findings and the results of the above-stated investigations. The types of chest X-ray findings of these 14 patients were fungus ball type (FB) in 2 patients, productive aspergilloma on the inner wall of a cavity (PAIC) in 8, mixed type with FB and PAIC in 2 and non-specific change in 2. However, there were 3 patients with the chest X-ray findings suggestive of PAIC in 7 patients (non-PA group) who were not diagnosed as pulmonary aspergillosis. Sputum culture were positive in 11 patients of PA group (79%) and negative in all patients of non-PA group. Precipitin antibody were positive in 8 patients of PA group (67%) and negative in all patients of non-PA group. PASTOREX ASPERGILLUS in serum were negative in all 21 patients. PASTOREX ASPERGILLUS in sputum were 3+ (+3) in all 14 patients of PA group and in 2 patients of non-PA group and 2+ (+2) in 2 patients of non-PA group. 12 patients of PA group were treated by antifungal agents and 11 patients responded well to the treatment. We also made sputum culture of Aspergillus and measurement of PASTOREX ASPERGILLUS in sputum on 14 patients (group B) who had respiratory diseases with stable cavities and 17 patients (group C) who had respiratory diseases without a cavity. In group B, sputum culture were positive in 1 patient and PASTOREX ASPERGILLUS in sputum were 3+ (+3) in 2 patients, 2+ (+2) in 1 and 1+ (+) in 2. In group C, sputum culture were negative in all patients and PASTOREX ASPERGILLUS in sputum were 3+ (+3) in 1 patient, 2+ (+2) in 3 and 1+ (+) in 2. Measurement of PASTOREX in sputum was a useful tool to diagnose non-invasive aspergillosis in addition to sputum culture and measurement of precipitin antibody titer. The sensitivity of PASTOREX in sputum was high but its specificity was low, however, its specificity could be raised by applying semi-quantitative analysis of PASTOREX in sputum.

[A clinical experience of DNA probe method for identifying Mycobacterium avium and Mycobacterium intracellulare].

In Japan the number of patient infected with M. avium complex (MAC) has been increasing in contrast to a decrease of pulmonary tuberculosis. DNA probe method enabled us be able to differentiate an isolated MAC into M. avium and M. intracellulare. From 1991 to 1992, we performed an investigation to apply this new technique of the DNA probe method on 52 patients of atypical mycobacteriosis diagnosed as infected with MAC by the ordinary method at the Higashi Nagoya National Hospital. The group consisted of 27 males and 25 females. M. avium infection was found in 39 patients (M. avium group) and M. intracellulare in 16 patients (M. intracellulare group). No significant gender difference was found between two groups. The M. avium group showed more complications in contrast to the M. intracellulare group. As complications in the M. avium group, pulmonary aspergillus infection, bacterial pneumonia and bronchiectasis were found in 4, 3 and 2 cases, respectively. The rate of drug resistance to antituberculosis drugs was high in the both groups. Chemotherapy with isoniazid (INH) rifampin (RFP) and streptomycin (SM) in five patients, that with INH, RFP and ethambutol (EB) in three were found to be effective after 4 months treatment. Three patients in M. avium group died of respiratory failure, aspergillus infection and renal failure. In contrast the prognosis of patients in the M. intracellulare group seemed to be better as there was no fatal case. We conclude that DNA probe method is useful to differentiate between M. avium and M. intracellulare, and enable us to select more appropriate selection of the chemotherapy and to assess of the prognosis.

[Clinical evaluation of plasma atrial natriuretic polypeptide in patients with respiratory failure in pulmonary tuberculosis].

We previously reported the clinical role of plasma immunoreactive atrial natriuretic polypeptide (ANP) and cyclic GMP in patients with respiratory diseases, bronchial asthma (BA), chronic pulmonary emphysema (CPE) and pulmonary insufficiency induced by pulmonary tuberculosis (TBC). In this study, moreover, we divided patients with respiratory failure induced by tuberculosis sequelae into two groups, patients with oxygen therapy group [O2 (+) group] or ordinary practical treatment group [O2 (-) group], and we evaluated the difference of the roles of ANP in two groups and the correlation of ANP and c-GMP with clinical findings, blood gas analysis, electrocardiogram, chest roentogen photography and spirogram in two groups. In conclusion, the respiratory failure in patients with tuberculosis sequelae is compensated by increased cardiac output, and that causes the rising of right atrial pressure. These results show, addition to the basic effects of ANP, the concentration of plasma ANP is released with relating the degree of respiratory failure.

[In vitro susceptibilities of Mycobacterium avium and Mycobacterium intracellulare to new macrolides, new quinolones, and antituberculous drugs on Dubos agar medium].

Mycobacterium avium and M. intracellulare were isolated from the sputum of patients infected with atypical mycobacteria using 1% Ogawa medium and identified by the DNA probe test. Then the MICs of various kinds of drugs against these mycobacterial species were determined on Dubos agar medium, and the drug susceptibilities were also determined on 1% Ogawa medium in parallel. The drugs tested were new macrolides, such as clarithromycin (CAM) and roxithromycin (RXM), new quinolones, such as ofloxacin (OFLX) and ciprofloxacin (CPFX), and antituberculous drugs, such as isoniazid (INH), rifampicin (PFP), streptomycin (SM), and ethambutol (EB). The MICs of the drugs tested, especially those of CAM, OFLX, and RFP, when determined on Dubos agar medium, were generally lower against M. intracellulare than against M. avium. The susceptibilities of the mycobacterial isolates tested to RFP and SM determined on Dubos agar medium were markedly different from those determined on 1% Ogawa medium. Such discrepancies may be accounted for by absorption of these drugs to the egg medium and instability of RFP in the egg medium. Overall, our results indicate that the new macrolides and new quinolones are effective against atypical mycobacteria.

The distribution of homologous enterococcal plasmid DNA sequences in human faecal isolates.

Hybridization was used to investigate the distribution of enterococcal plasmid sequences among 306 strains of Enterococcus and Streptococcus spp. isolated from faeces of humans of various ages. As DNA probes for the survey three plasmids, whose DNAs did not hybridize each other and designated as pMS13, pTW34 and pHK30, were selected from plasmids borne in Ent. faecalis. pTW34 DNA hybridized only with DNAs from enterococci, with high frequency in Ent. faecalis and low frequency in Ent. faecium. pMS13 DNA hybridized with DNAs of all Enterococcus spp. tested and with Strep. bovis, Strep. equinus and Strep. salivarius. Eighty-five percent of Ent. faecium isolates had sequences homologous to pMS13 but in the other species the values were less than 60%. Some enterococci had DNAs which hybridized with the pHK30 probe. The different distribution of the three DNA sequences indicates the possibility that plasmid DNAs encode advantageous phenotypes for the colonization of bacteria in the lumen of the bowel.

[Preoperative treatment of esophageal cancer with oral peplomycin jelly].

Pepleo-jelly, peplomycin (PEP) compounded with sodium polyacrylate (PANa), was used as a local preoperative chemotherapy for esophageal cancer. Thirty milligrams of PEP emulsified in 10 ml of jelly was orally administered for 17 patients on consecutive bed time, and the average total dose was 470 mg. Concentrations of PEP in blood and tissue were measured with a bioassay. PEP was not detected in the blood but was detected in esophageal tissue and regional lymphnodes. A significant difference (p less than 0.05) of tissue concentrations between normal esophageal mucosa (0.11 microgram/g) and malignant region (0.34 microgram/g) was noted. Furthermore, PEP was detected in many regional lymphnodes. Histological antitumor effect (Ef2) was noted in three of the eight metastatic lymphnodes. A side effect of this treatment was not recognized. It is believed that this treatment is safe and useful in combination with other adjuvant cancer therapy.

[Bypass operation adaptable to stature increase in child with atypical coarctation of the aorta].

A 9-year-old girl was admitted with hypertension and severe congestive heart failure. Upon physical examination, a discrepancy of blood pressure between arm and leg was noted. Aortography revealed narrowing about 5 cm in length at the midportion of the descending thoracic aorta. Bypass operation of the narrow segment was performed under mild hypothermia with the diagnosis of atypical coarctation of the aorta. It was supposed that the patient might outgrow the graft and the graft would become too small for grown-up patient in diameter and length, then the haemodynamics would become less satisfactory and too much tension on the suture line would occur. A woven Dacron graft, 10 mm in diameter, 15 cm in length, was anastomosed proximally and distally to the coarcted segment at a distance of about 6 cm. So, the graft was disposed in a C-shaped configuration. It was expected that the arch of the graft would open more widely with increase of her stature, even if the graft does not increase in length. She has been followed for twelve years. Hypertension of upper extremity and arm-to-leg gradients of the systolic blood pressure were recognized from two years after the operation, particularly with exercise. However, cardiomegaly and left ventricular hypertrophy in ECG were improved. She appears to have been developing normally with no cardiac symptoms. Estimating from angiography, the distance between proximal and distal anastomoses stretched about 2 cm during the period of rapid growth, though calcification of the graft had been seen from four years after the operation, perhaps due to increased calcium turnover in childhood.