Frequency of failure to isolate Shigella spp. by the direct plating technique and improvement of isolation by enrichment in selenite broth.

In order to clarify the failure to isolate Shigella spp. by direct plating, we compared frequencies of Shigella spp. isolation by direct plating and by plating after enrichment in selenite broth. A total 67 strains were isolated in this study. The strains of 38 (56.7%) were isolated only by direct plating, and 25 (37.3%) strains were isolated by both direct plating and after enrichment. Four strains (6.0%) were isolated after enrichment but not by direct plating. Since 6% of isolated Shigella spp. were not isolated by direct plating, we recommend that direct plating and additional isolations from selenite broth should be performed. The significance concerning reduction of concentration of sodium selenite in enrichment broth is discussed.

[Effect of serotype specific glycopeptidolipid (GPL) isolated from Mycobacterium avium complex (MAC) on phagocytosis and phagosome-lysosome fusion of human peripheral blood monocytes].

Mycobacterium avium complex (MAC) is a typical intracellular parasite similar to M. tuberculosis and is one of the most important pathogens that coinfects AIDS patients. Attention has been focused on M. avium infection causing immunosuppression of hosts. Specific serotype-subspecies such as 1, -4 or -8 serotypes can be isolated frequently in humans infected with HIV. Furthermore, the prognosis after infection differs depending on the serotype. Serotype-4 in general shows unfavourable prognosis, while serotype-16 yields rapid recovery. Therefore, we have been interested in the immunomodifying activity of the surface glycopeptidolipid (GPL) antigen. However, no information has been available to date dealing on the virulent factor of MAC that is directly related with intracellular bactericidal activity. Recently, we have tried to test the effect of various GPLs purified from MAC on phagocytic processes of human peripheral blood monocytes (PBMC). We have used GPL-coated heat-killed staphylococcal cells to be phagocytosed by PBMC, and phagosome-lysosome fusion (P-L fusion) was estimated by the acridine orange staining of fused vesicles and bacteria. Results showed strong promotion of phagocytosis and marked inhibition of P-L fusion by serotype-4 GPL, while neither promotion of phagocytosis nor inhibition of P-L fusion in phagocytic cells were shown by serotype-16 GPL. Serotype-8 GPL showed concomitant stimulation of both phagocytosis and P-L fusion. These effects may be due to some unknown interaction between specific carbohydrate chain and organella membranes and serotype-4 GPL may be one of the possible virulent factors in MAC. Comparison with known possible virulent factors such as trehalose 6,6'-dimycolate (TDM), trehalose 6-monomycolate (TMM), glucose 6-monomycolate (GM) or sulfatide was also reported.

[Bacteriological studies of traveller's diarrhoea (6). Analysis of enteropathogenic bacteria at Kansai Airport Quarantine Station from September 4th, 1994 through December 1996].

During the period of investigation from Sept. 4, 1994 to Dec, 1996, a total of 11,446,534 overseas travellers were quarantined at Kansai Airport Quarantine Station, and 22,187 voluntarily reported of episodes suffering from diarrhoea. Bacteriological examination of the stools a total of 9,299 individuals' was performed, and the following results were obtained. 1) Various enteropathogenic bacteria were isolated from 33.3% of the stools examined. Bacterial species isolated were as follows: Plesiomonas shigelloides, 2,066 cases (66.7%); Vibrio parahaemolyticus, 358 cases (11.6%); Aeromonas sobria, 360 cases (11.6%); Shigella spp., 291 cases (9.4%); Salmonella spp., 183 cases (5.9%); A. hydrophila, 126 cases (4.1%); and V. cholerae non-O1, 121 cases (3.9%). However, ETEC was not done with an object of test. 2) In 502 cases (16.2%), plural enteropathogenic bacteria were isolated from single patient, suggesting high frequency of a mixed infections. 3) From Feb. to Mar. 1995, thirteen cases cholera were found from patients who had travelled to Bali, Indonesia. Cases with enteropathogenic bacteria other than V. cholerae O1 were found without any seasonal variation. 4) The major regions where the travellers were infected with the pathogens are as follows: Vibrio spp., were from only Asia; Shigella, widely distributed but especially in India and Indonesia; P. shigelloides and Salmonella, widely distributed. 5) Among the Shigella strains, S. sonnei were isolated the most, followed by S. flexneri, S. boydii and S. dysenteriae. A strain of S. boydii provisional serovar E 16553 was isolated from a patient infected in India. 6) Among the Salmonella serovars, Salmonella Enteritidis was isolated the most frequently (49 cases, 25.7%). 7) 265 (89.2%) of 297 Shigella strains, 52 (27.2%) of 19] Salmonella strains, and 19 (95.0%) of 20 V. cholerae O1 were resistant to one or more drugs tested (SM. CP. TC. KM. ABPC. NA. OFLX). 8) All of the 20 V. cholerae O1 strains were Ogawa, E1 Tor. All of them were toxigenic strains. 9) The most frequently isolated serovar of V. parahaemolyticus was O3: K6. 89.8% of all V. parahaemolyticus strains were positive for thermostable direct hemolysin (TDH) gene, and 14.6% of them were positive for TDH-related hemolysin (TRH) gene by DNA-probe or PCR method.

[Studies on bacillary dysentery cases of overseas travellers--during 1979 to 1995].

A total of 36,780,440 overseas travellers during 1979-1995 (17 years) were quarantined at Osaka and Kansai Airport Quarantine Station, 84,777 travellers reported themselves suffer from diarrhoea. Stools from 29,587 persons were bacteriologically examined. Various enteropathogenic bacteria were isolated from 9,766 (33.0%) patients of the stools examined. Isolated species were as follows: Plesiomonas shigelloides (3,234 cases); Salmonella spp. (2,236 cases); enterotoxgenic Escherichia coli (1,621 cases); Vibrio parahaemolyticus (1,959 cases); and Shigella spp. (1,242 cases). 1,278 different Shigella strains were isolated from 1,242 cases who were thus diagnosed as bacillary dysentery patients. The suspected regions or countries for infection of these cases were analysed. The serovars and antibiotic-sensitivities of the isolated strains were examined. Colicine typing of S. sonnei strains were also done. The results can be summarized as follows. 1) The most cases (53.4%) were infected in India. 2) The percentage distribution of sub-species of the strains was as follows; S. sonnei (57.8%), S flexneri (29.8%), S. boydii (8.4%), and S. dysenteriae (4.0%), respectively. 3) The major colicine type of S. sonnei strains were type 6 and 0. 4) The percentage of Antibiotic-resistant strains of each sub-species was S. dysenteriae (92.2%), S. sonnei (89.4%), S. flexneri (87.1%), and S. boydii (84.9%), respectively. The percentage of Antibiotic-resistant strains of S. flexneri were increased annually.

[Incidence of Kanagawa phenomenon-positive and -negative Vibrio parahaemolyticus strains isolated from traveller's diarrhea and their relation to tdh and trh genes].

A total of 1,319 strains of Vibrio parahaemolyticus isolated from traveller's diarrhea were analysed for Kanagawa phenomenon (KP) with the Wagatsuma blood agar test and the results were also compared with those of analyses of tdh and trh genes which encode thermostable direct hemolysin (tdh) and TDH-related hemolysin (trh). The majority of the strains (1,152 strains) counting 87.3% had positive KP, among which 1,049 and 103 strains were only tdh and both tdh and trh-positive ones, respectively. However, 167 strains counting 12.7%, which is quite high compared to the previous report, were found to have negative KP, among which 94 and 24 strains were only trh and both tdh and trh-positive ones, respectively.

[Bacteriological studies of travellar's diarrhoea. 5) Analysis of enteropathogenic bacteria at Osaka Airport Quarantine Station from January 1992 through September 3rd, 1994].

During the last 2 years and 8 months before the closure of Osaka Airport Quarantine Station (from Jan. 1992 to Sep. 3, 1994), a total of 7,421,909 overseas travellers were quarantined. 15,919 reported themselves of suffering from diarrhoea. Bacteriological examination of a total of 6,031 individuals' stools were performed. 1) Various enteropathogenic bacteria were isolated from 31.2% of the stools examined. Isolated species were as follows: Plesiomonas shigelloides, 1,127 cases (59.9%); Vibrio parahaemolyticus, 293 cases (15.6%); Salmonella spp., 262 cases (13.9%); Shigella spp., 235 cases (12.5%); Aeromonas sobria, 93 cases (4.9%); V. cholerae non-O1, 69 cases (3.7%). 2) The enteropathogenic bacteria were isolated through out the year without any seasonal variation. 3) The major regions where the travellers were infected with the pathogens are as follows: V. cholerae non-O1 (NAG Vibrio) and enteropathogenic Escherichia coli, South-East and South-West Asia; Vibrio other than NAG, South-East and East Asia; Shigella, widely distributed but especially in India; P. shigelloides and Salmonella, widely distributed. 4) 2 strains of toxigenic (cholera toxin-producing) V. cholerae O139 were isolated from patients who had visited Indonesia and Thailand, respectively. 5) In 320 cases (17%), plural enteropathogenic bacteria were isolated from single patients, suggesting a high frequency of the mixed infections. 6) Among Shigella strains, S. sonnei were isolated the most, followed by S. flexneri (24.7%), S. boydii (8.8%) and S. dysenteriae (2.9%). 7) Among Salmonella serovers, Salmonella Enteritidis was isolated the most frequently (39 cases, 14.1%). 8) 218 (91.6%) of 238 Shigella strains and 103 (37.6%) of 276 Salmonella strains were resistant to one or more drugs tested (SM.CP.TC.KM.ABPC.NA.OFLX). 9) All of the 22 V. cholerae O1 strains were Ogawa, E1 Tor. Among them, 19 were toxigenic strains and 3 were non-toxigenic. 10) O4:K8 was the most frequently isolated serover of V. paraemolyticus. 87.4% of all V. parahaemolyticus strains were positive with thermostable direct hemolysin (TDH) gene, and 12.6% of them were positive with TDH-related hemolysin (TRH) gene by DNA-probe methods.

[A case of hydronephrosis caused by renal stones with elevated serum levels of CA-19-9 and CA-125].

A case of hydronephrosis caused by renal stones with elevated serum levels of carbohydrate antigens 19-9 (CA-19-9) and 125 (CA-125) is reported. A 74-year-old woman was hospitalized with pyelonephritis. The results of computerized tomography and endoscopy suggested that the patient did not have pancreatic or ovarian cancer, but laboratory tests were significant for serum CA-19-9 greater than 1,000 U/ml (normal less than 37) and serum CA-125 78 U/ml (normal less than 35), which are markers for such tumors. Abdominal X-ray films showed stones in the pelvis of the right kidney. Excretory urography showed that the right kidney was not functioning. Computerized tomography showed severe right hydronephrosis with a thin renal cortex. Complete obstruction of the ureteropelvic junction of the kidney was seen by retrograde pyelography. During percutaneous pyelography, aspiration of the pelvic urine was done, and the sample had a high concentration of CA-19-9 (250,000 U/ml), but no cancer cells. The patients underwent right nephrectomy. The pelvic urine was tested for both CA-19-9 and CA-125 this time, and both were high (190,000 U/ml and 5,100 U/ml, respectively). Pathological evaluation showed no evidence of a malignant tumor. The epithelium lining the renal pelvis was stained for both CA-19-9 and CA-125 using the avidin-biotin-peroxidase complex method. Serum levels of the markers returned to normal after nephrectomy.

Multicenter study with recombinant human erythropoietin.

A multicenter study with recombinant human erythropoietin (rh-EPO) was carried out. Of 172 hemodialysis patients with anemia selected for the study from 20 hospitals and clinics, 77 were males and 95 females (mean age 53.9 years). A starting dose of 1,500 U of rh-EPO (Epoetin beta) was administered intravenously at the end of every dialysis session. If the efficacy was not acceptable, the dose was increased to 3,000 U. When the target hematocrit was achieved (30%), the total dose was decreased. The results of the study were excellent relative to those of other multicenter studies with regard to efficacy, safety, and changes in laboratory data. The incidence of hypertension was lower in our study compared with other reports because we used a low initial dose. The efficacy of rh-EPO therapy was determined earlier and more reliably by reticulocytes than by hematocrit or hemoglobin. Prompt iron supplement therapy is recommended with careful observation of serum iron and ferritin.

[Application of new intraurethral stent for higher risk patients with benign prostatic hypertrophy].

We reviewed our experience of the application of a new urethral stent (PROSTAKATH) for 8 patients with prostatic outflow obstruction from December 1989 to March 1990. 6 patients had required the Foley catheter for several months because of chronic urinary retention. 2 were dys-uric patients having a higher bladder residual urine volume. All were in a high risk group for surgery. 7 out of the 8 patients were treated successfully. The stent was not placed in one. All 7 patients in whom the urethral stent was placed voided freely after placement of the stent. Bladder residual urine was not detected by the ultrasound sonography except in one patient. The urethral stent used in this study was a spring-like spiral with an outside diameter of 21 Fr, it is made of gold-plated stainless steel. Under local anesthesia, it can be easily inserted using a 6-7 Fr. ureteral catheter as a guide wire under ultrasonic scanning guidance. During the follow-up period of 5-8 months, 1 patient had an episode of migration of the stent to the bladder 2 months later, which was removed endoscopically, and a new stent was placed. Side effects were observed in 2 patients; one complained of strong discomfort and the other suffered from urge incontinence. Both symptoms were ameliorated during the follow-up period. We conclude that the urethral stent is an effective device as a non-invasive treatment of prostatic outflow obstruction.

Stone fragility--measurement of stone mineral content by dual photon absorptiometry.

We measured the mineral content of urinary tract stones by dual photon absorptiometry, which is widely used for the analysis of bone mineral content, and compared the values of the stones by dual photon absorptiometry (DPA values) with the results of an in vitro fracture study as well as those of an in vivo extracorporeal shock wave lithotripsy treatment study. The results of a preliminary experiment showed that the DPA values of 20 urinary tract stones reflected actual stone mineral content. As a result of the in vitro fracture study, the DPA value calculated by volume of a struvite stone, which was the most easily disintegrated, was the lowest (0.53 g/cm3). The DPA values of calcium oxalate monohydrate and apatite stones, which poorly disintegrated, were the highest (0.98, 1.01 g/cm3). The DPA value of calcium oxalate dihydrate, which moderately disintegrated, was 0.86 g/cm3. By the in vivo extracorporeal shock wave lithotripsy treatment study, the total DPA values of stones measured before extracorporeal shock wave lithotripsy treatment in 12 patients were 0.73 +/- 0.34 g in successful cases and 1.92 +/- 0.43 g in unsuccessful cases with a significant difference between the two (p less than 0.05). These results showed that the measurement of stone mineral content by dual photon absorptiometry was useful in predicting the fragility of stones against shock waves before performing extracorporeal shock wave lithotripsy treatment for patients with urinary tract stones.