J Clin Microbiol, 1998 Jun, 36(6), 1544 - 8 Identification of new influenza B virus variants by multiplex reverse transcription-PCR and the heteroduplex mobility assay; Zou S et al.; A quick genetic approach for the screening of influenza virus variants was developed in this laboratory (S . Zou, J . Clin . Microbiol . 35:2623-2627, 1997) . It uses multiplex reverse transcription and multiplex PCR to amplify and differentiate the variable region of the hemagglutinin genes of different types and subtypes of influenza viruses . Variants within the same type or subtype are then identified by the heteroduplex mobility shift assay of the amplicons . The method was used to screen influenza virus isolates received from provincial laboratories during the 1996-1997 season and was able to identify new influenza B virus variants . Sequencing of the amplicons derived from the hemagglutinin gene of the identified variants and comparison with the vaccine strain B/Harbin/7/94 showed substitution rates of 2.26 to 2.55% at the nucleotide level and 4.26 to 4.68% at the amino acid level . The result further demonstrated that the approach provides a quick, sensitive, and reliable screening for influenza virus variants . It also suggested the necessity of close monitoring of influenza B virus isolates in the 1997-1998 season and critical evaluation of the reference strain for the type B influenza virus. J Clin Microbiol, 1998 Jun, 36(6), 1530 - 3 Routine use of PCR-reverse cross-blot hybridization assay for rapid identification of Mycobacterium species growing in liquid media; Sanguinetti M et al.; A PCR-reverse cross-blot hybridization assay procedure that is able to rapidly identify 13 species of clinically relevant mycobacteria was evaluated for routine use in the identification of acid-fast isolates growing in BACTEC 460 TB (12B and 13A) and BACTEC 9000 MB (Myco/F) liquid media . Eight of the probes used were already described by Kox et al . (L . F . F . Kox et al., J . Clin . Microbiol . 33:3225-3233, 1995) . In addition, we used six other probes specific for M . chelonae, M . malmoense or M . szulgai, M . genavense, M . gordonae, M . terrae, and M . marinum/M . ulcerans that we designed ourselves . This procedure allowed us to identify 459 mycobacterial species directly from broth cultures of 5,466 clinical samples collected over 1 year and processed with the radiometric or nonradiometric BACTEC system . Our results were in agreement with those obtained by conventional identification methods and also with those obtained by mycolic acid analysis by high-performance liquid chromatography . This assay seems to be a reliable procedure for the routine identification of mycobacteria, providing an accurate identification of mycobacterial isolates more rapidly than conventional tests, with remarkable implications for an efficacious specific antimycobacterial therapy. Clin Lab Manage Rev, 1998 Jul-Aug, 12(4), 248 - 55 Impact on human resources: Core Laboratory versus laboratory information system versus modular robotics; Dadoun R; Technological advances in laboratory systems have had a great impact on human resources . Surviving the changes requires an in-depth understanding of the technology to implement the appropriate operational model . St . Mary's is a 414-bed, acute care hospital . For 18 months, the laboratories went through the process of moving from a noncomputerized traditional model laboratory (i.e., by discipline) to a fully computerized Core Lab . The Core Lab concept fully integrates biochemistry, hematology, blood bank, and microbiology into two sections (not physically separated): tests processed by automation and tests processed manually . This approach led to a 15% reduction in staff while the volume doubled . The transitions occurred sequentially: from traditional laboratory to Core Lab (noncomputerized), from manual Core Lab to fully computerized Core Lab, and ultimately from a simulation of manual preanalytical phase to automated preanalytical phase (modular robotics) . The findings show that Core Lab and computerization have almost the same impact on human resources, 35% and 30% respectively, and modular robotics the least impact with 17%. Methods, 1998 May, 15(1), 15 - 26 Biochemical methods for analysis of kinetoplastid RNA editing; Sabatini RS et al.; RNA editing is a posttranscriptional process involving mRNAs {reviewed by K . Stuart et al . (1997) Microbiol . Mol . Biol . Rev . 61, 105-120; G . J . Arts and R . Benne (1996) Biochim . Biophys . Acta 1307, 39-54; and S . L . Hajduk and R . S . Sabatini (1996) in Molecular Biology of Parasitic Protozoa (Smith, D . S., and Parsons, M., Eds.), pp . 134-158, Oxford Univ . Press, Oxford} and tRNAs {K . M . Lonergan and M . Gray (1993) Science 259, 812-816} that has now been described in an increasing number of eukaryotic organisms . In this process sequences differ from their gene sequences by the addition, removal, or conversion of specific ribonucleotides . RNA editing was first described within the mitochondrion of kinetoplastid protozoa . Several of the mitochondrial mRNAs in these flagellates have uridine residues inserted and deleted at specific sites . In some cases, more than 50% of the mRNA is created by RNA editing . In this article, we describe some of the biochemical methods used in analyzing the process of RNA editing in kinetoplastid mitochondria. Ann Rheum Dis, 1998 Feb, 57(2), 107 - 9 Quality control of synovial fluid crystal identification; von Essen R et al.; OBJECTIVE: To establish a quality assessment programme for the diagnosis of crystal arthropathies by synovial fluid (SF) microscopy . METHODS: Three or four cytocentrifuge slides prepared from suitable patient SF specimens were distributed to 25-47 predominantly Finnish clinical laboratories once a year . Sodium urate crystals were included in every survey . RESULTS: Returns for the years 1989-1996 were reviewed . Laboratories that participated in > four surveys made on an average one error a year (range 0.25-2) . The error rate for specimens containing abundant crystals was acceptable but it increased considerably for specimens showing few crystals per microscope field . No laboratory characteristic predictive of successful performance was found . CONCLUSION: Errors in quality assessment results for crystal identification were much more frequent than in the fields of, for example, clinical chemistry or microbiology . Despite efforts to provide educational feedback, no improvement was seen during the study period . Because of the dearth of data from other parts of the world it is not known for certain whether this study has merely pinpointed a local problem or if the same trend applies elsewhere. Stomatologiia (Mosk), 1994 Apr-Jun, 73(2), 22 - 6 {The multivariate analysis of clinico-laboratory data in stomatology}; Balashov AN et al.; The authors demonstrate the potentialities of multidimensional analysis methods: component, cluster, and discriminant as exemplified by clinico-laboratory examinations of 26 patients with medium-severe periodontitis and concomitant diseases . A two-stage modification of multidimensional grouping of biomedical objects is suggested . The first stage implies detection of the principal components and their estimation for each examinee . The second stage consists in realization of multidimensional grouping method according to G . S . Kil'dishev and Iu . I . Abolentsev in relation to individual component values . Different typologically homogeneous groups of patients with a specific microbiologic and biochemical status evidently requiring a differentiated approach to treatment may be singled out within the frames of a current classification of periodontal diseases basing on clinical and laboratory data. J Comput Assist Tomogr, 1998 May-Jun, 22(3), 437 - 43 Osteomyelitis of the pelvis/hips in paralyzed patients: accuracy and clinical utility of MRI; Huang AB et al.; PURPOSE: The goal of our study was to determine the accuracy and clinical utility of MRI in the diagnosis of osteomyelitis of the pelvis/hips in paralyzed patients . METHOD: In 44 paralyzed patients, 59 consecutive MR examinations of the pelvis/hips were evaluated prospectively . Criteria for diagnosis of osteomyelitis were based on those established in previous studies of complex, nonhematogenous osteomyelitis (diabetic foot) . Average follow-up was 3 years . The standard of reference for the diagnosis of osteomyelitis was histologic/microbiologic results of surgical biopsy specimens or clinical follow-up . Note was made if decubitus ulcers, sinus tract, fistula, fluid collection, abscess, septic arthritis, joint effusion, bursitis, or heterotopic ossification was present on MRI . Comparison of the extent of infection by MRI and surgical margins was performed . Effect on surgical treatment was defined by absence of recurrent infection at the surgical site within 6 weeks of limited resection . RESULTS: The criteria for diagnosis of osteomyelitis were fulfilled in 49 of 57 MR studies for an overall accuracy of 97% . There was one false-negative MR study . MRI for the diagnosis of osteomyelitis yielded a sensitivity of 98% and a specificity of 89% . There were 41 decubitus ulcers, 28 sinus tracts, 2 fistulae, 14 fluid collections, 15 abscesses, 9 hips with septic arthritis, 10 "bland" hip effusions, 5 cases of trochanteric bursitis, and 30 patients with heterotopic ossification . Twenty-one patients underwent limited surgical resection guided by MR findings in which only the enhancing area was resected . There was only one recurrence of osteomyelitis at the surgical margins . CONCLUSION: MRI is accurate in the diagnosis of osteomyelitis and associated soft tissue abnormalities in spinal cord-injured patients . MRI can delineate the extent of infection in guiding limited surgical resection and preserving viable tissue. Eur J Obstet Gynecol Reprod Biol, 1998 May, 78(1), 5 - 10 Elevated interleukin-8 concentrations in amniotic fluid of mothers whose neonates subsequently develop bronchopulmonary dysplasia; Ghezzi F et al.; OBJECTIVE: To determine if an intrauterine sub-clinical inflammatory process is a risk factor for the development of bronchopulmonary dysplasia . METHODS: A cohort study was conducted in patients who met the following criteria: (1) Singleton gestation; (2) preterm labor or preterm premature rupture of the membranes; (3) amniocentesis for microbiologic studies of the amniotic fluid and (4) delivery between 24 and 28 weeks of gestation . Bronchopulmonary dysplasia was defined as the need for supplemental oxygen for 28 days or longer after birth, associated with compatible chest radiographic findings . Amniotic fluid interleukin-8, was measured using a specific immunoassay . Logistic regression analysis and bootstrap procedure were used for statistical purposes . RESULTS: Forty-seven patients met the inclusion criteria for this study . Among these patients, the prevalence of bronchopulmonary dysplasia was 23.4% (11/47) . Amniotic fluid culture was positive in 21 out of 47 (44.7%) patients . Median (range) amniotic fluid interleukin-8 concentration was higher in patients whose neonates subsequently developed bronchopulmonary dysplasia than in those who did not (17 {9.8-583.7} ng ml(-1) versus 9.6 {0.91-744} ng ml(-1), P=0.057) . An amniotic fluid IL-8 level greater than 11.5 ng ml(-1) was far more common in mothers whose fetuses went on to develop bronchopulmonary dysplasia than in those who did not (10/11 {90.9%} versus 17/36 {47%}; P=0.01) . This relationship remained significant even after correcting for the effect of gestational age and birthweight (Odds ratio: 11.9; P<0.05) . CONCLUSION: Sub-clinical intrauterine inflammation is a risk factor for the subsequent development of bronchopulmonary dysplasia . We propose that in utero aspiration of fluid with high concentration of pro-inflammatory mediators may contribute to the lung injury responsible for the development of bronchopulmonary dysplasia. J Clin Periodontol, 1998 Apr, 25(4), 271 - 7 Interrelationship between periodontics and adult orthodontics; Ong MA et al.; The purpose of this review article is to provide the dental practitioner with an understanding of the interrelationship between periodontics and orthodontics in adults . Specific areas reviewed are how periodontal tissue reacts to orthodontic forces, influence of tooth movement on the periodontium, effect of circumferential supracrestal fiberotomy in preventing orthodontic relapse, effect of orthodontic bands on the periodontium, specific microbiology associated with orthodontic bands, mucogingival considerations and time relationship between orthodontic and periodontal therapy . In addition, the relationship between orthodontics and implant restorations (e.g., using dental implants as orthodontic anchorage) will be discussed. Telemed J, 1998 Spring, 4(1), 11 - 7 Telemicrobiology: feasibility study; McLaughlin WJ et al.; BACKGROUND: Rural hospitals generally lack staffing with infectious disease specialists or pathologists . Without on-site pathologists, the range of microbiology services offered by clinical laboratories may be limited as well . OBJECTIVE: To study the feasibility of using static-image telepathology to evaluate Gram stains of microbiologic preparations . MATERIALS AND METHODS: In this retrospective feasibility study, three pathologists evaluated Gram stains of slides from 50 cases by two viewing modalities: static-image telepathology and conventional light microscopy . Digital video images of slides were captured at two magnifications (using 40x and 100x objective lenses) at 1024 x 768 x 24-bit color and transmitted over standard telephone lines at 14,400 kbps . Pathology reports and culture results served as "truth diagnoses." Categories of interpretations were correct, minor discrepancy, or major discrepancy with regard to the implications for patient care . RESULTS: The diagnostic accuracy of video image readings and conventional light microscopy readings were nearly identical, with no statistically significant differences in the performances of specialty and nonspecialty pathologists (P > 0.05) . The mean accuracies of readings of the video images and light microscopy images were 95.3% and 95.4%, respectively . Taking into account the time required by a referring pathologist to capture video digital images, telemicrobiology was somewhat less efficient than conventional light microscopy . CONCLUSIONS: Pathologists can accurately evaluate digital video images of preselected fields on Gram-stained slides . In clinical practice, however, a limiting factor may be the availability of local personnel qualified to select the microscopic fields for evaluation by telepathologists . The adequacy of the video images suggests that telepathology may also be used for remote supervision of quality assurance programs in microbiology laboratories, as well as for remote proficiency training of laboratory personnel. J Clin Microbiol, 1998 Apr, 36(4), 979 - 82 Performance of a PCR assay for detection of Pneumocystis carinii from respiratory specimens; Caliendo AM et al.; This study evaluates the performance of a PCR assay for the detection of Pneumocystis carinii from respiratory specimens that has been designed for use in the clinical microbiology laboratory . The test includes a simple method for nucleic acid extraction and amplification, a colorimetric probe hybridization technique for detection of amplicons, and an internal control to evaluate for the presence of inhibitors of amplification . Two hundred thirty-two clinical specimens (120 induced-sputum {IS} and 112 bronchoalveolar lavage {BAL} specimens) from 168 patients were tested by both immunofluorescent (direct fluorescent-antibody {DFA}) staining and PCR . Of the 112 BAL specimens, 17 were positive for P . carinii by DFA staining and PCR . An additional two specimens were DFA negative and PCR positive . For BAL specimens, the sensitivity and specificity of PCR compared to DFA were 100 and 98%, respectively . Eighteen IS specimens were positive for P . carinii by DFA, and 27 were positive by PCR . One of the 18 DFA-positive IS specimens was negative by PCR; this patient had just completed therapy for P . carinii pneumonia . Of the 10 specimens that were PCR positive and DFA negative, 4 were from patients who had a subsequent BAL specimen that was positive by DFA and PCR . For IS specimens, the sensitivity of DFA and PCR was 82 and 95%, respectively . The specificity of PCR for IS specimens was 94% . Due to the high sensitivity of PCR for the detection of P . carinii from IS specimens, a PCR-based diagnostic test may be a useful screening test and may alleviate the need for bronchoscopy in some patients. Chest, 1998 May, 113(5), 1394 - 8 The role of atropine premedication in fiberoptic bronchoscopy using intravenous midazolam sedation; Williams T et al.; OBJECTIVE: Atropine premedication is widely used for fiberoptic bronchoscopy and may help by drying secretions, producing bronchodilatation, or preventing vasovagal reactions . The objective of this study was to see whether atropine premedication is really of practical benefit when patients are sedated with i.v . midazolam . DESIGN: In a double-blind study, patients were randomly allocated to receive i.m . atropine (0.6 mg) or saline placebo (1 mL) as premedication 30 to 60 minutes before they were sedated with progressive doses of i.v . midazolam until judged to be lightly asleep . SETTING: A District General Hospital in England . PARTICIPANTS: One hundred consecutive patients referred for bronchoscopy . MEASUREMENTS AND RESULTS: Samples taken during the procedure were washings for microbiology and cytology and brushings for cytology and biopsy, but no transbronchial biopsies . Peak flow readings were recorded before premedication and before the start of the procedure . During the procedure an estimate was made of pharyngeal and tracheobronchial secretions, bleeding, use of saline to wash out secretions, and local anesthetic needed to control coughing . Patients were monitored for saturation and cardiac rhythm . There was no significant bronchodilatation after premedication in either group, nor were there differences in secretions, use of saline, tracheobronchial bleeding, desaturation, and arrhythmias . More local anesthetic was needed to control coughing in the placebo group (mean 357 mg vs 331 mg in the atropine group, p=0.02), but this was not of practical significance . CONCLUSION: When intravenous midazolam sedation is used for bronchoscopy, atropine premedication is not of benefit. Epidemiol Infect, 1998 Mar, 120(2), 117 - 23 Invasive pneumococcal infection in South and West England; Smith MD et al.; Variation in the incidence of invasive pneumococcal disease across South and West England, in 1995, was measured through a survey of microbiology laboratories . A 100% response rate was achieved . The incidence by laboratory varied between 5.2 and 20.4 per 100,000 catchment population (P < 0.001) . Adjusting for pneumococcal vaccine uptake rate in over 65 year olds, hospital admission rates, blood culture system used and for the age and sex structure of the population, did not account for this variation . When blood culture sampling rates were included in a logistic regression model, the variation between laboratories was much less and of lower statistical significance (P = 0.019) . Higher rates of blood culture sampling were associated with a higher incidence of invasive pneumococcal disease . Consistently high sampling should be encouraged because a higher diagnostic rate should result in more selective prescribing of antibiotics, and secondly because improved ascertainment of severe pneumococcal infections is a prerequisite for the evaluation of new pneumococcal conjugate vaccines. Sheng Li Ke Xue Jin Zhan, 1996 Apr, 27(2), 101 - 6 {Helicobacter pylori: a great discovery}; Wang ZJ; Within the past decade, there has been an explosion of investigative activity and publications about Helicobacter pylori pathogen . Its role in gastroduodenal diseases is becoming remarkably evident . It is accepted that H . pylori is intimately involved in the pathogenesis of chronic gastritis microbiol/DT and peptic ulcer disease . Furthermore, it is also associated with stomach cancers . The old creed in medical field that "No acid, no ulcer" would probably be changed to "No H . pylori infection, no ulcer" . So, the discovery of H . pylori revolutionized our understanding of gastroduodenal diseases. Biophys J, 1998 May, 74(5), 2272 - 7 Spatial sensing of stimulus gradients can be superior to temporal sensing for free-swimming bacteria; Dusenbery DB; Predictions of the minimal size an organism must have to swim along stimulus gradients were used to compare the relative advantages of sensory systems employing spatial (simultaneous) and temporal (sequential) gradient detection mechanisms for small free-swimming bacteria, leading to the following conclusions: 1) there are environmental conditions where spatial detection mechanisms can function for smaller organisms than can temporal mechanisms, 2) temporal mechanisms are superior (have a smaller size limit) for the difficult conditions of low concentration and shallow gradients, but 3) observed bacterial chemotaxis occurs mostly under conditions where spatial mechanisms have a smaller size limit, and 4) relevant conditions in the natural environment favor temporal mechanisms in some cases and spatial mechanisms in others . Thus, sensory ecology considerations do not preclude free-swimming bacteria from employing spatial detection mechanisms, as has been thought, and microbiologists should be on the lookout for them . If spatial mechanisms do not occur, the explanation should be sought elsewhere. Otolaryngol Pol, 1998, 52(1), 5 - 10 {Deep neck infections in the antibiotic era}; Zietek E et al.; The paper presents 43 patients treated between 1971 and 1995 in the Otolaryngology Clinic of the Pomeranian Medical Academy in Szczecin because of deep neck space infections . The authors analyse their own clinical material paying particular attention to the diagnosis, microbiology, patients prognosis and basic therapeutic procedures . Out of the 43 cases, deaths occurred in 3, which denotes 7% mortality rate . The reason for failure was discussed . It was revealed that needle aspiration and sonography were for valuable to identify early abscess formation . The analyses have confirmed that the prognosis of the deep neck space infections is influenced by early diagnosis and prompt, extensive surgical treatment. Hinyokika Kiyo, 1998 Mar, 44(3), 193 - 4 {A case of septic shock following incarceration of the penis}; Horiguchi A et al.; An 84-year-old male presented to the emergency room with the chief complaint of painful, swollen penis following the use of a constriction ring to maintain penile erection . A high fever, chills and hypotension were recognized . Septic shock was presumed, and administration of antibiotics was started . Microbiologic cultures revealed Escherichia coli in blood . We herein report a rare but serious complication accompanying incarceration of the penis. Eur J Cardiothorac Surg, 1998 Feb, 13(2), 190 - 5 Frozen section diagnosis and surgical biopsy of lymph nodes, tumors and pseudotumors of the mediastinum; de Montpreville VT et al.; OBJECTIVE: Our experience with the use of frozen section (FS) was reviewed in order to assess its interest and limits, during minimally aggressive mediastinal surgery for staging of lung carcinomas and biopsy of primary lesions . METHODS: The retrospective study was based on a series of 420 consecutive mediastinal biopsies with FS performed through cervicotomy (n = 12), mediastinoscopy (n = 345), mediastinotomy (n = 43), manubriotomy (n = 13) or videothoracoscopy (n = 7), in 417 patients, aged 8-86 years (mean: 52.5 +/- 15.6) . The FS diagnoses were compared with the final diagnoses and consequences of the FS analyses were analyzed . RESULTS: Biopsies with FS had a 99.4% (351/353) efficiency rate for a precise definitive diagnosis (excluding normal lymph nodes) . However, two patients required a second operation: one for typing a lymphoma and the other to correct a FS misdiagnosis of small cell carcinoma on a crushed normal lymph node . The sensitivity for detection of carcinoma lymph node metastases was 99% (200/202) . The two false negative FS diagnoses, including one post-chemotherapy, were related to micrometastases . There were 46 correct FS diagnoses of non-metastatic lymph node which were followed by an immediate thoracotomy during the same anesthesia, for benign lung lesions (n = 9) or for carcinomas (n = 37), including 30 lung carcinomas that were immediately resected and proved to have no mediastinal involvement (n = 24), or only a limited involvement in a non-biopsied site (n = 6) . In the 51 cases of primary mediastinal tumors excluding carcinomas, FS indicated a resectable lesion with a sensitivity of 87.5% (7/8) and a specificity of 97.7% (43/44) . Five lesions were immediately resected: one Castleman's disease, one intrathoracic goiter, two of six thymomas and a Hodgkin's disease, which was diagnosed as thymoma on FS . An invasive thymoma was resected during a second operation after a FS diagnosis of carcinoma . FS had a sensitivity of 100% in the 62 cases of sarcoidosis and a sensitivity of 90% in the 20 cases of infectious lesions . One of the 18 cases of tuberculosis and an infectious pseudo-tumor of the anterior mediastinum had no microbiologic study because of FS diagnoses of tumor necrosis and lymphoma . CONCLUSIONS: FS efficiently secures the adequacy of the samples and guides the surgeon's decision making for the resection of lung carcinomas, but is less effective for a precise diagnosis of some primary mediastinal lesions, which may have close histologic appearances. J Marmara Univ Dent Fac, 1993 Sep, 1(4), 337 - 41 Dental procedures and infective endocarditis: current approaches of cardio-vascular surgery departments from different centres (Part 1); Akincibay H et al.; The relationship between dental procedures and infective endocarditis is well known . Therefore, in order to prevent infective endocarditis, pre-operative antibiotic prophylaxis is recommended . However, a total agreement has not been achieved regarding which dental procedures and specific heart diseases require prophylaxis . The aim of the present study is to review the latest concepts dealing with heart diseases regarding the choice of appropriate antibiotic regimen, determination of patients at risk of infective endocarditis and dental procedures which require pre-operative prophylaxis . Our review revealed that there is still some disagreement regarding the above concepts . We believe that dentists, especially periodontists, can play a significant role in the prevention of infective endocarditis following dental procedures, by giving more attention to the subject and also by alerting the infectious diseases/microbiology departments and related medical staff. Genitourin Med, 1997 Dec, 73(6), 498 - 502 Evaluation of the microparticle enzyme immunoassay Abbott IMx Select Chlamydia and the importance of urethral site sampling to detect Chlamydia trachomatis in women; Brokenshire MK et al.; OBJECTIVE: To evaluate the commercial microparticle enzyme immunoassay (MEIA), Abbott IMx Select Chlamydia, for the detection of Chlamydia trachomatis in women and to compare its performance with endocervical cell culture . Also, to determine whether sampling the urethral site is an important part of chlamydial diagnosis in women . SETTING: The Auckland, Manukau, and Waitakere Sexual Health Clinics, Auckland, New Zealand and the Department of Clinical Microbiology, Auckland Hospital, Auckland, New Zealand . PATIENTS: The study population consisted of 622 consecutive women who attended the three sexual health clinics . METHODS: The IMx Chlamydia assay was performed on an IMx analyser, following a specimen treatment procedure . All reactive samples from the IMx Chlamydia assay were confirmed using the IMx Chlamydia blocking antibody reagent . The Syva direct fluorescent antibody (DFA) test was used to aid in resolving discrepancies . The cell culture technique was performed in shell vials using cycloheximide treated McCoy cells, which were stained using a fluorescein conjugated monoclonal antibody . RESULTS: When compared against the endocervical cell culture, the IMx Chlamydia had a sensitivity of 82.1% (23/28) and a specificity of 99.3% (590/594) . When compared against an expanded gold standard, the IMx Chlamydia and endocervical cell culture had sensitivities of 84.4% (27/32) and 87.5% (28/32), specificities of 100% (590/590) and 100% (590/590), positive predictive values of 100% (27/27) and 100% (28/28), negative predictive values of 99.2% (590/595) and 99.3% (590/594), and accuracies of 99.2% (617/622) and 99.4% (618/622), respectively . The prevalence rate by endocervical cell culture and the expanded gold standard were 4.5% and 5.1%, respectively . Additional urethral cell culture testing revealed a further nine patients positive from this site only, giving a 28% (9/32) increase in the number of patients diagnosed for chlamydia, thus giving an overall prevalence of 6.6% (41/622) . CONCLUSIONS: The IMx Chlamydia assay is an easy and rapid test to perform, it is cost effective, and shows similar performance to endocervical cell culture in the female population studied and is thus an excellent alternative to culture for the diagnosis of C trachomatis . The study also showed the importance of urethral site sampling in these women, as endocervical testing alone will underestimate the prevalence of chlamydial genital infection. Enferm Infecc Microbiol Clin, 1998 Jan, 16(1), 14 - 8 {Meningitis by enterovirus in pediatrics . Clinical characteristics and virologic diagnosis}; Rubio G et al.; BACKGROUND AND METHODS: With the aim of knowing the etiology of aseptic meningitis (pleocytosis in LCR with negative bacterial culture and positive or negative virus culture or without pleocytosis with viral culture in positive LCR), 48 children attending the Emergency Department of our hospital from June to December, 1995 were prospectively studied . Pharyngeal and rectal swab, LCR for bacteria and virus and blood cultures were carried out . The samples were inoculated in fibroblasts MRC-5, RD and BGM . The cytopathic effect was identified by immunofluorescence . Typing was performed in the National Center of Public Health Care Microbiology, Virology and Immunology in Majadahonda (Spain) . RESULTS: Isolation of the virus was positive in 40/48 (83.3%) of the children: in 17 (35.5%) of LCR and the remaining 23/40 (47.9%) of pharyngeal and/or rectal swab . In all the cases the cytopathic effect was detected in MRC-5 . The viruses found were echovirus 30 and 5 . A greater incidence of the disease was observed in November and December . CONCLUSIONS: Meningitis by enterovirus is frequent in our area . Culture of the virus in LCR, the pharynx and stools is useful on suspicion of aseptic meningitis since the virus may be isolated from LCR in more than one third of the patients . Serotyping aids in surveilling the appearance of outbreaks and to know the predominant viruses . Cell culture is the diagnostic treatment of choice, but has a low sensitivity and is slow . The application of other techniques such as PCR which have a greater sensitivity and with high specificity for the diagnosis of these infections is necessary. Anesthesiology, 1998 Apr, 88(4), 935 - 44 Open-lung biopsy in patients with acute respiratory distress syndrome; Papazian L et al.; BACKGROUND: It has been suggested that fibrosis present during the fibroproliferative phase of acute respiratory distress syndrome (ARDS) can be treated by corticosteroids . However, neither clinical nor microbiologic criteria permit differentiation of this fibroproliferative phase from a nosocomial pneumonia . The aim of this observational case series was to evaluate the safety and utility of open-lung biopsy (OLB) performed in patients receiving ventilatory support who had persistent ARDS despite negative bacterial cultures . METHODS: During a 4-yr period, 37 OLBs were performed in 36 of 197 patients receiving ventilatory support who had ARDS . The severity of ARDS was assessed by a lung injury score of 3.1 +/- 0.4 (mean +/- SD) and a median ratio of the partial pressure of oxygen (PaO2) to the fraction of inspired oxygen (FiO2) of 118 mmHg . Histologic examination; bacterial, fungal, and acid-fast staining; and cultures of the tissue sample were performed . RESULTS: Fibrosis was present in only 41% of the lung specimens obtained by OLB . Only six patients received corticosteroids (17%) . In 9 of the 15 patients with fibrosis, cytomegalovirus pneumonia precluded the use of corticosteroids . Histologic cytomegalovirus pneumonia was diagnosed in 18 cases . Histologic bacterial or mycobacterial pneumonia was diagnosed in five cases . No significant change in arterial blood gases was noted as linked to the biopsy procedure except an increase of the PaO2/FiO2 ratio . One pneumothorax was diagnosed on a chest roentgenogram 12 h after OLB . Only one patient required blood transfusion during the 48-h period after OLB (for an hemothorax) . Five patients had moderate air leaks from operative chest tubes for 2-10 days . CONCLUSIONS: Open lung biopsy appeared to be a useful and acceptably safe diagnostic technique in patients with ARDS . It permitted the diagnosis of unexpected cytomegalovirus pneumonia. Am J Obstet Gynecol, 1998 Apr, 178(4), 759 - 64 Inhibition of amniotic prostaglandin E release by ampicillin; Vesce F et al.; OBJECTIVE: The effect of antibiotics in the prevention of preterm labor needs to be further investigated . The aim of this study was to determine the effect of ampicillin on prostaglandin E release from amnion as a possible explanation for its ability to retard preterm labor . STUDY DESIGN: The effect of the beta-lactam antibiotic ampicillin on prostaglandin E release from human amnion was tested under basal and stimulated conditions . RESULTS: Ampicillin dose dependently inhibits basal prostaglandin E release from amnion in both static and dynamic conditions . In our experiments, 10(-7) mol/L ampicillin (a concentration able to significantly inhibit prostaglandin E output) leaves the microbiologic features of the medium substantially unmodified up to 5 hours of incubation . Moreover, the drug reversibly counteracts the prostaglandin E elevation induced by arachidonic acid or oxytocin . CONCLUSION: This finding (i.e., that ampicillin inhibits prostaglandin E release from amnion) may offer an explanation for a beneficial response to ampicillin therapy in the case of preterm labor even in the absence of bacterial infection. Minerva Stomatol, 1998 Jan-Feb, 47(1-2), 51 - 6 {Aspergillus-induced maxillary sinusitis}; Fancello A et al.; Maxillary aspergillosis sinusitis more often occur in immunocompromised patients, but this pathology is increasing also in healthy people . Many risk factors are known and clinical and radiologic findings are aspecific; diagnosis therefore is possible with histologic and microbiologic aid . This particular disease and its surgical-pharmacological approach is explained. Eur J Biochem, 1998 Apr 1, 253(1), 263 - 9 The function of the periplasmic Sud protein in polysulfide respiration of Wolinella succinogenes; Klimmek O et al.; The periplasmic Sud protein was previously isolated as a sulfide dehydrogenase from Wolinella succinogenes . Sud modified by a C-terminal His-tag (Sud-His6) was produced in Escherichia coli by expression of the sud gene . Sud-His6 catalyzed thiocyanate formation from cyanide and polysulfide . The Vmax of this activity was more than one order of magnitude higher than that of sulfide oxidation by dimethyl-naphthoquinone and that of polysulfide reduction by BH4- . The apparent Km was less than 20 microM polysulfide . Polysulfide and not elemental sulfur was found to be the product of sulfide oxidation by dimethyl-naphthoquinone, in contrast to the earlier view {Kreis-Kleinschmidt, V., Fahrenholz, F., Kojro, E . & Kroger . A . (1995) Arch . Microbiol . 165, 65-68} . Sud-His6 did not contain metal ions or other prosthetic groups . Replacement by site-directed mutagenesis of the single cysteine residue of the Sud monomer caused complete loss of activity, while the exchange of the single histidine residue or of the lysine residue situated next to cysteine did not affect activity . In equilibrium dialysis, the Sud-His6 monomer bound up to ten polysulfide sulfur atoms with a dissociation constant of 0.2 mM . Sud-His6 loaded with polysulfide sulfur showed an absorption spectrum in the range of 350-400 nm; this spectrum differed from that of free polysulfide . Electron transport from H2 to polysulfide catalyzed by the membrane fraction of W . succinogenes was stimulated by the presence of small amounts of Sud-His6 . The apparent Km for polysulfide decreased sevenfold in the presence of saturating amounts of Sud-His6 (1 microM Sud-His6 dimer) . Similar results were obtained with intact W . succinogenes cells containing low and high amounts of Sud . Sud appears to function as a polysulfide binding protein and probably binds polysulfide sulfur to its cysteine residue and transfers it to the substrate site of the membraneous polysulfide reductase. Appl Environ Microbiol, 1998 May, 64(5), 1805 - 11 Use of inducible feedback-resistant N-acetylglutamate synthetase (argA) genes for enhanced arginine biosynthesis by genetically engineered Escherichia coli K-12 strains; Rajagopal BS et al.; The goal of this work was to construct Escherichia coli strains capable of enhanced arginine production . The arginine biosynthetic capacity of previously engineered E . coli strains with a derepressed arginine regulon was limited by the availability of endogenous ornithine (M . Tuchman, B . S . Rajagopal, M . T . McCann, and M . H . Malamy, Appl . Environ . Microbiol . 63:33-38, 1997) . Ornithine biosynthesis is limited due to feedback inhibition by arginine of N-acetylglutamate synthetase (NAGS), the product of the argA gene and the first enzyme in the pathway of arginine biosynthesis in E . coli . To circumvent this inhibition, the argA genes from E . coli mutants with feedback-resistant (fbr) NAGS were cloned into plasmids that contain "arg boxes," which titrate the ArgR repressor protein, with or without the E . coli carAB genes encoding carbamyl phosphate synthetase and the argI gene for ornithine transcarbamylase . The free arginine production rates of "arg-derepressed" E . coli cells overexpressing plasmid-encoded carAB, argI, and fbr argA genes were 3- to 15-fold higher than that of an equivalent system overexpressing feedback-sensitive wild-type (wt) argA . The expression system with fbr argA produced 7- to 35-fold more arginine than a system overexpressing carAB and argI genes on a plasmid in a strain with a wt argA gene on the chromosome . The arginine biosynthetic capacity of arg-derepressed DH5 alpha strains with plasmids containing only the fbr argA gene was similar to that of cells with plasmids also containing the carAB and argI genes . Plasmids containing wt or fbr argA were stably maintained under normal growth conditions for at least 18 generations . DNA sequencing identified different point mutations in each of the fbr argA mutants, specifically H15Y, Y19C, S54N, R58H, G287S, and Q432R. J Clin Microbiol, 1998 May, 36(5), 1236 - 9 Detection of Ureaplasma urealyticum in endotracheal tube aspirates from neonates by PCR; Nelson S et al.; A PCR-based test was optimized for the detection of Ureaplasma urealyticum from neonatal respiratory specimens, with primers directed against the multiple-banded antigen gene (L . J . Teng, X . Zheng, J . I . Glass, H . Watson, J . Tsai, and G . H . Cassell, J . Clin . Microbiol . 32:1464-1469, 1994) . Endotracheal tube aspirates (225) from 103 low-birth-weight neonates (<1,250 g) were taken, when possible, at days 0, 4, and 14 after birth and examined by culture and by PCR . Of 77 specimens positive by either method, 73 were detected by PCR and 60 were detected by culture . Overall, 36% of the neonates were positive for U . urealyticum by either method . Of 16 patients with PCR-positive-culture-negative results, 13 had positive cultures at another sampling point, and one additional patient had a twin with positive cultures . Of 11 patients with day 0 specimens positive by PCR alone, 9 subsequently became culture positive, demonstrating the utility of this test in early detection . Multiple serovars were present in over 50% of positive specimens, with serovars 3 and 14 in combination being most prevalent . The amplicon size generated from the specimen by PCR correctly predicted the biovars isolated in over 85% of positive specimens . Thus, this PCR test was valuable in allowing early detection of U . urealyticum in neonatal respiratory specimens, as well as in providing biovar information. J Bacteriol, 1998 May, 180(9), 2515 - 21 Developmental regulation of transcription of whiE, a locus specifying the polyketide spore pigment in Streptomyces coelicolor A3 (2) Kelemen GH, Brian P, Flardh K, Chamberlin L, Chater KF, Buttner MJ. whiE is a complex locus that specifies the polyketide spore pigment in Streptomyces coelicolor A3(2) . Two divergently oriented promoters, whiEP1 and whiEP2, were identified in the whiE gene cluster, and their activities were analyzed during colony development in wild-type and sporulation-deficient strains . Both promoters were developmentally regulated; whiEP1 and whiEP2 transcripts were detected transiently at approximately the time when sporulation septa were observed in the aerial hyphae, and transcription from both promoters depended on each of the six known "early" whi genes required for sporulation septum formation (whiA, -B, -G, -H, -I, and -J) . Mutation of the late sporulation-specific sigma factor gene, sigF, had no effect on the activity of whiEP1 but blocked transcription from whiEP2 . However, sigmaF-containing holoenzyme was not sufficient to direct transcription of whiEP2 in vitro . The whiEP2 promoter controls expression of whiE ORFVIII, encoding a putative flavin adenine dinucleotide-dependent hydroxylase that catalyzes a late tailoring step in the spore pigment biosynthetic pathway . Disruption of whiE ORFVIII causes a change in spore color, from grey to greenish (T.-W . Yu and D . A . Hopwood, Microbiology 141:2779-2791, 1995) . Consistent with these observations, construction of a sigF null mutant of S . coelicolor M145 caused the same change in spore color, showing that disruption of sigF in S . coelicolor changes the nature of the spore pigment rather than preventing its synthesis altogether. Appl Environ Microbiol, 1998 May, 64(5), 1766 - 72 Molecular analysis of a laccase gene from the white rot fungus Pycnoporus cinnabarinus; Eggert C et al.; It was recently shown that the white rot basidiomycete Pycnoporus cinnabarinus secretes an unusual set of phenoloxidases when it is grown under conditions that stimulate ligninolysis (C . Eggert, U . Temp, and K.-E . L . Eriksson, Appl . Environ . Microbiol . 62:1151-1158, 1996) . In this report we describe the results of a cloning and structural analysis of the laccase-encoding gene (lcc3-1) expressed by P . cinnabarinus during growth under xylidine-induced conditions . The coding region of the genomic laccase sequence, which is preceded by the eukaryotic promoter elements TATA and CAATA, spans more than 2,390 bp . The corresponding laccase cDNA was identical to the genomic sequence except for 10 introns that were 50 to 60 bp long . A sequence analysis indicated that the P . cinnabarinus lcc3-1 product has a Phe residue at a position likely to influence the reduction-oxidation potential of the enzyme's type 1 copper center . The P . cinnabarinus lcc3-1 sequence was most similar to the sequence encoding a laccase from Coriolus hirsutus (level of similarity, 84%). Appl Environ Microbiol, 1998 May, 64(5), 1743 - 9 Sensitive and rapid detection of viable Giardia cysts and Cryptosporidium parvum oocysts in large-volume water samples with wound fiberglass cartridge filters and reverse transcription-PCR; Kaucner C et al.; We recently described a reverse transcription-PCR (RT-PCR) for detecting low numbers of viable Cryptosporidium parvum oocysts spiked into clarified environmental water concentrates . We have now modified the assay for direct analysis of primary sample concentrates with simultaneous detection of viable C . parvum oocysts, Giardia cysts, and a novel type of internal positive control (IPC) . The IPC was designed to assess both efficiency of mRNA isolation and potential RT-PCR inhibition . Sensitivity testing showed that low numbers of organisms, in the range of a single viable cyst and oocyst, could be detected when spiked into 100-microliter packed pellet volumes of concentrates from creek and river water samples . The RT-PCR was compared with an immunofluorescence (IF) assay by analyzing 29 nonspiked environmental water samples . Sample volumes of 20 to 1,500 liters were concentrated with a wound fiberglass cartridge filter . Frequency of detection for viable Giardia cysts increased from 24% by IF microscopy to 69% by RT-PCR . Viable C . parvum oocysts were detected only once by RT-PCR (3%) in contrast to detection of viable Cryptosporidium spp . in four samples by IF microscopy (14%), suggesting that Cryptosporidium species other than C . parvum were present in the water . This combination of the large-volume sampling method with RT-PCR represents a significant advance in terms of protozoan pathogen monitoring and in the wider application of PCR technology to this field of microbiology. Acta Cytol, 1998 Mar-Apr, 42(2), 305 - 11 Relative value and cost-effectiveness of culture and special stains in fine needle aspirates of the lung; Krane JF et al.; OBJECTIVE: To define the role of microbiologic stains and culture in lung fine needle aspiration (FNA) specimens . STUDY DESIGN: All lung FNA specimens over a nine-year period, with results of both culture and microbiologic stains (Gram's, methenamine silver and acid fast) were reviewed and correlated with clinical information . RESULTS: Sixty-five cases were identified; 13 cases represented clinically significant infections (20%) . Gram's stain identified 3 infections and had 1 false positive result, while culture identified 7 infections and had 9 false positive results . However, all false positive cultures represented easily identifiable contaminants, and eight of nine cases had no associated acute inflammation or necrosis . Aspergillus species were detected in four cases by Papanicolaou and silver stain, while culture was positive in only one case . Coccidioides immitis was detected by both Papanicolaou stain and culture in one case . A single case of Mycobacterium tuberculosis was identified by both culture and acid-fast stain . While culture appeared more cost-effective than Gram's stain for identifying bacteria, both Papanicolaou and methenamine silver stain were more cost-effective for identifying fungi . CONCLUSION: In lung FNA specimens, culture and special stains should be restricted to specimens with acute inflammation or necrosis . Gram's stain and fungal culture are insensitive and not cost-effective, and fungi are often identifiable with routine stains. Ned Tijdschr Geneeskd, 1998 Feb 21, 142(8), 415 - 9 {Hepatic steatosis during treatment with zidovudine and lamivudine in an HIV-positive patient}; ter Hofstede HJ et al.; A 33-year-old HIV-infected man was given antiretroviral therapy with zidovudine and lamivudine . After ten months' treatment the patient had elevated hepatic transaminase levels . Severe hepatic steatosis was found in the biopsy . Clinical history, laboratory, microbiologic and X-ray examination revealed no other abnormalities . The transaminase levels remained high after withdrawal of zidovudine alone, but a decrease was observed when both zidovudine and lamivudine were stopped . Rechallenge of lamivudine therapy caused the levels to increase again . The hepatic steatosis was considered to be caused by the antiretroviral therapy, lamivudine having a synergistic influence on this side effect of zidovudine . Ten months after the therapy was changed to the protease inhibitor indinavir combined with zalcitabine and stavudine, two other nucleoside analogues, hepatic steatosis recurred. Crit Care Clin, 1998 Apr, 14(2), 221 - 50 Postcardiac surgery infections; Lutwick LI et al.; Infections following cardiac surgery, although generally uncommon, are associated with difficult management decisions and significant morbidity and mortality . They often present while the patient is either in a critical care unit, or requires CCU management . This review analyzes infections related to median sternotomy wounds, prosthetic heart valves, transvenous permanent pacemakers, automatic implantable cardioverter-defibrillators, and left ventricular assist devices . The diagnosis, microbiology, treatment and outcome of each is also discussed. Cesk Patol, 1998 Jan, 34(1), 3 - 6 Evolution of pathology practice in the United States: current status and future trends; Sims KL et al.; This article traces the development of pathology practice from its origins in autopsy pathology to its current practice in the United States . The American model of practice differs markedly from that in continental European countries because of the extensive incorporation of "Clinical Pathology" with the traditional disciplines of anatomic pathology under the auspices of the Pathology Department . "Clinical Pathology" as it is now defined includes the laboratory testing disciplines of Chemistry, Hematology, Immunology, Medical Microbiology, and Transfusion Medicine . The increasing importance of computers and information management, DNA diagnostic techniques, and the multiple roles of the pathologist as a researcher and consultant in pathology practice in the United States is discussed. Dent Today, 1998 Feb, 17(2), 104, 106, 108 - 11 The key to success: maintaining the long-term health of implants; Silverstein L et al.; During the infancy years of dental implantology, the emphasis for long-term success of osseointegrated implants was the surgical phase of dental implantology . In the years that followed, the emphasis for success had switched from a purely surgical influence to focusing more on the proper fixture placement which would be dictated by the prosthetic and aesthetic needs of each particular case . In more recent years, the dental profession has recognized professional implant maintenance and diligent patient home care as two critical factors for the long-term success of dental implants . The microbiota and clinical presentation of peri-implantitis is the same as periodontitis around a natural tooth. Microsc Res Tech, 1998 Mar 15, 40(6), 463 - 72 Cytochemical and biochemical studies on adenylate cyclase activity in preneoplastic and neoplastic liver tissue and cultured liver cells; Mayer D et al.; The ATP-analogue adenylyl(beta,gamma-methylene)diphosphonate was chosen as substrate for the cytochemical localization of adenylate cyclase (AC) activity . The tissues investigated covered normal rat liver and liver from carcinogen-treated animals with preneoplastic lesions and hepatocellular neoplasms, as well as cultured liver cells . The AC reaction product methylene diphosphonate was precipitated with Pb2+ immediately at the place of production . This approach permitted a precise localization of AC activity by light and electron microscopy . The specificity of the AC reaction was demonstrated by control reactions, including inhibition of AC with 2'5'-dideoxyadenosine and activation with forskolin, glucagon, and cholera toxin . Endogenous phosphatases were inhibited with tetramisole and NAD . In normal liver, AC activity was mainly localized in the sinusoidal membrane of hepatocytes . A distinct gradient in activity was observed within the liver lobule . Hepatocytes localized around the terminal hepatic venule showed a significant higher AC activity compared to hepatocytes near the portal tract . AC was clearly decreased in focal preneoplastic liver lesions of the glycogenotic-basophilic cell lineage leading to hepatocellular carcinomas . Cytochemically detected intensity of AC activity corresponded to data obtained by microbiochemical assays in laser-dissected tissue samples . A remarkable interdependence of AC activity and degree of differentiation was also seen in epithelial rat liver cell lines: Highly differentiated cells show high enzyme activity and vice versa, as shown by both cytochemical and biochemical examinations . It is concluded that alterations in cellular signal transduction caused by alterations in AC activity play an important role in hepatocarcinogenesis. J Pediatr Ophthalmol Strabismus, 1998 Mar-Apr, 35(2), 73 - 9; quiz 110-1 Clinical practice guidelines for the management of orbital cellulitis; Uzcategui N et al.; PURPOSE: A retrospective study was performed to review the management of periorbital and orbital cellulitis at Miami Children's Hospital, between January 1, 1993 and February 15, 1996 . RESULTS: One hundred and one patients were included in this study . The variables analyzed in this study included age, sex, length of hospital stay, imaging studies, laboratory tests, and microbiology specimens collected . Patients were classified according to the modified Chandler classification . Average length of stay for our patient population was 4.5 days . Data regarding the radiologic studies, laboratory tests, and microbiology yield of specimens were analyzed . CONCLUSIONS: The incidence of orbital infection, manifested by lid swelling alone is much more common (stages I and II) than orbital infection involving postseptal findings (stages III, IV, and V); 84.16% compared with 15.84%, respectively . An updated approach and a general guideline for the management of periorbital and orbital cellulitis according to the clinical staging of the process is presented. J Virol, 1998 May, 72(5), 4508 - 14 Feline coronavirus type II strains 79-1683 and 79-1146 originate from a double recombination between feline coronavirus type I and canine coronavirus; Herrewegh AA et al.; Recent evidence suggests that the type II feline coronavirus (FCoV) strains 79-1146 and 79-1683 have arisen from a homologous RNA recombination event between FCoV type I and canine coronavirus (CCV) . In both cases, the template switch apparently took place between the S and M genes, giving rise to recombinant viruses which encode a CCV-like S protein and the M, N, 7a, and 7b proteins of FCoV type I (K . Motowaka, T . Hohdatsu, H . Hashimoto, and H . Koyama, Microbiol . Immunol . 40:425-433, 1996; H . Vennema, A . Poland, K . Floyd Hawkins, and N . C . Pedersen, Feline Pract . 23:40-44, 1995) . In the present study, we have looked for additional FCoV-CCV recombination sites . Four regions in the pol gene were selected for comparative sequence analysis of the type II FCoV strains 79-1683 and 79-1146, the type I FCoV strains TN406 and UCD1, the CCV strain K378, and the TGEV strain Purdue . Our data show that the type II FCoVs have arisen from double recombination events: additional crossover sites were mapped in the ORF1ab frameshifting region of strain 79-1683 and in the 5' half of ORF1b of strain 79-1146. Oral Microbiol Immunol, 1996 Oct, 11(5), 365 - 8 Characterization of a 4.2-kb plasmid isolated from periodontopathic spirochetes; Chan EC et al.; Oral anaerobic treponemes are assoicated with active periodontal disease and may comprise up to 57% of the microbiota in periodontal pockets . Four treponeme strains (designated U2a, U2b, U9b, and U9c) isolated from clincial cases were found to harbor a new 4.2-kb plasmid when plasmid DNA was extracted and purified employing the Qiagen Plasmid Kit . This plasmid differs from the smaller plasmids (2.0-, 2.6-, and 2.7-Kb) reported previously by others in Treponema denticola . The newly discovered 4.2-kb plasmid was found to be the same in all four treponeme strains by restriction endonuclease analysis . It is a circular plasmid since restriction with PstI, Pvu II, Sma I, Xma I, Ava 1 or Bam HI produced a single band of the same size . Bacterial strain U2b was shown to be Treponema socranskii and U9c to be T . denticola . The plasmid is designated "pTS1" . The presence of the same plasmid in different species of the treponemes isolated from the same patient suggests the possibility of a naturally occurring genetic transfer system within the oral spirochetes or their ability to take up and maintain mobilizable plasmids. Acta Microbiol Immunol Hung, 1997, 44(4), 309 - 25 Extended-spectrum beta-lactamases: an actual problem of hospital microbiology (a review); Szabo D et al.; Although there is a variety of mechanisms of bacterial resistance to beta-lactam antibiotics, the most important one is production of beta-lactamases inactivating penicillins and cephalosporins . The classification of beta-lactamases is based on biochemical, enzymological (i.e . molecular structure, inhibitory property, substrate-profile, relative rate of hydrolysis) and immunological characters . Extended-spectrum beta-lactamases (ESBLs) can be derived from TEM or SHV enzymes . These enzymes have now been sequenced and it has been found that relatively few point mutations have occurred in the gene of the TEM and SHV type enzymes . These point mutations clustered in five areas of the gene . The amino acid mutations can alter the conformation, the active site and change the hydrance of beta-lactamase-cephalosporin binding capacity . So the enzyme is able to bind and hydrolyse the third generation cephalosporins . Successive mutation interacted radically increasing the binding capacity of enzymes and confer resistance to newer cephalosporins . The use of these drugs provides a strong selective pressure to develop these mutations . Sporadic nosocomial outbreaks due to strains producing an ESBL led to an epidemic problem in some hospitals resulting in a concurrent dissemination of genes, plasmids or strains . Clinical epidemiological importance and role of ESBLs and emergence of multiply resistance of bacteria of nosocomial importance are discussed in this brief. Rev Med Brux, 1998 Feb, 19(1), 5 - 9 {Septic thrombophlebitis of the internal jugular vein and Lemierre syndrome}; Bouton F et al.; The authors report a case of Lemierre's syndrome . This uncommon clinical entity is characterized by a septic internal jugular vein thrombosis with secondary metastatic abscesses and Fusobacterium necrophorum septicemia, following an acute oropharyngeal infection . The diagnosis is primarily clinical and it should be suspected when a severe septicaemic illness, with pulmonary symptoms, occurs after an acute pharyngotonsillar infection . This article reviews the clinical picture, microbiology and treatment of this forgotten complication of acute tonsillitis. Rev Esp Salud Publica, 1997 Mar-Apr, 71(2), 181 - 7 {The evaluation of epidemiological services related to brucellosis in the Autonomous Community of the Basque Country}; Coll Jorda D et al.; BACKGROUND: Spain's Epidemiologic Notifiable Disease Surveillance System (ENDSS) was evaluated by reference to Brucellosis, the zoonoses with greatest incidence countrywide and a disease that is notifiable on a case by case basis in the Basque Autonomous Region (BAR) . METHODS: ENDSS information and the use to which it was put, were analysed . Active case searching was carried out via the Microbiologic Information System (MIS), public hospitals and the Occupational Disease Registry (ODR) . A comparison was run between reported and search data . The capture-recapture method was used to compare ENDSS against hospital cases . RESULTS: Risks and outbreaks wee detected . Suspect cases were not reported . There was a 40% post-search increase in cases, i.e., 40 versus a previous figure of 24 . Sensitivity was estimated at 60% . Predictive Value Positive could not be computed however . No case definition was in force . As estimated by the capture-recapture method, overall sensitivity for both ENDSS and hospital-based surveillance was 84% (95% IC: 70-100) . The number of cases estimated by this method was 38 +/- 8 . CONCLUSIONS: Underreporting is in evidence and, as a consequence, sensitivity is low . A case definition is called for . Response time is slowed by awaiting confirmation before reporting . The number of cases detected through active searching is compatible with that calculated by the capture-recapture method using only two sources. Ophthalmology, 1998 Apr, 105(4), 707 - 16 Additional procedures after the initial vitrectomy or tap-biopsy in the Endophthalmitis Vitrectomy Study; Doft BH et al.; OBJECTIVE: The study aimed to assess the frequency, indications, and outcome of additional ocular procedures after initial treatment of vitrectomy (VIT) or tap-biopsy (TAP) for patients with endophthalmitis after cataract extraction . DESIGN: The study design was an analysis of observational data collected as part of a multicenter, randomized clinical trial . PARTICIPANTS: Of the 420 patients enrolled in the Endophthalmitis Vitrectomy Study, the 148 who had additional procedures were compared with the 272 who did not . MAIN OUTCOME MEASURES: The types, indications, and number of additional ocular procedures were assessed . A masked examiner measured visual acuity 9 to 12 months after study entry . RESULTS: Within 1 week of study entry, 8% of VIT eyes and 13% of TAP eyes underwent additional procedures, 14% for complications of the initial procedure and 86% for worsening ocular inflammation or infection . Cultures were obtained in 33 of the 38 eyes operated on for worsening inflammation or infection and were positive in 42% . Cultures obtained from the early additional procedures were positive more frequently in eyes with an initial TAP (71%) than in eyes with an initial VIT (13%) . Both virulence of initial microbiologic organism isolated and poor presenting vision were risk factors for requirement of reoperation . In all cases in which a single organism was cultured at the initial procedure, when the reculture was positive, it was the same organism . Late additional procedures (after 7 days) were required in 27% of patients . Visual outcome was much worse for eyes that had an additional procedure compared to eyes that did not, and this was especially the case for eyes that had an early additional procedure . Only 15% of eyes that had an early additional procedure achieved 20/40 visual acuity as compared to 57% of eyes that did not . CONCLUSION: Need for an additional procedure was a marker of more severe disease, and patients who underwent additional procedures achieved poorer visual acuity at final follow-up. Am J Orthop, 1998 Mar, 27(3), 207 - 10 Relation of surface area to in vitro elution characteristics of vancomycin-impregnated polymethylmethacrylate spacers; Holtom PD et al.; To study the relationship between surface area and antibiotic elution from antibiotic-impregnated polymethylmethacrylate (PMMA) spacers, a standard block spacer made of vancomycin (4 g) and 40 g of PMMA was compared with two unique spacer designs, the "donut" and "fenestrated." The spacers were incubated in phosphate-buffered saline, which was changed daily, and a microbiologic assay was used to measure the antibiotic activity of the eluates . The donut and fenestrated spacers had 12% and 40% more surface area than the standard spacer, respectively . There was no significant difference, however, in daily elution levels of antibiotic between the donut spacer and the standard spacer . The fenestrated spacer displayed significantly better elution than either the standard or donut spacers, with an average of 20% more antibiotic eluted on any given day. Vet Pathol, 1998 Mar, 35(2), 108 - 16 Cutaneous and systemic necrotizing vasculitis in swine; Thibault S et al.; A systemic vasculitis involving particularly the skin and kidneys has been recently described in swine under the name dermatitis/nephropathy syndrome . Twelve pigs with gross cutaneous lesions typical of this condition were necropsied, and morphologic, immunohistochemical, microbiologic, and epidemiologic characteristics were studied . The pigs were divided into three groups comprising eight pigs with acute lesions, two with chronic lesions, and two with acute lesions kept for sequential skin biopsies . Acute skin lesions consisted of round to irregular, red to purple macules and papules that often coalesced to form large, irregular patches and plaques . With time, the lesions became covered by crusts and faded gradually, sometimes leaving scars . Characteristic distribution included the perineal area of the hindquarters, limbs, dependent parts of the abdomen and thorax, and margins of the ears . In the acute phase of the disease, necrotizing and leucocytoclastic vasculitis of small-caliber blood vessels were observed within the dermis and panniculus and in various extracutaneous locations such as the renal pelvis and synovial membranes . All pigs had macroscopic evidence of pneumonia and generalized lymphadenopathy . Microscopically, they had interstitial pneumonia and perivascular cuffing of mononuclear cells in various tissues including skin . The presence of immunoglobulins and complement was demonstrated by immunofluorescence in and around necrotic vessels of the skin in the early stages . Porcine reproductive and respiratory syndrome (PRRS) virus (PRRSV) antigens were detected by immunohistochemistry in macrophages located around vessels of the tissues examined (skin and kidneys) in acute and chronic cases . PRRSV RNA was demonstrated by reverse transcription-polymerase chain reaction in lung and spleen homogenates from all pigs . The PRRSV was isolated in cell culture from 11 of the pigs . These findings suggest that PRRSV infection may play a role in the pathogenesis of this systemic vascular disease of swine. J Biochem (Tokyo), 1998 Feb, 123(2), 247 - 52 Purification and characterization of a puromycin-hydrolyzing enzyme from blasticidin S-producing Streptomyces morookaensis; Nishimura M et al.; Blasticidin S-producing Streptomyces morookaensis JCM4673 produces an enzyme which inactivates puromycin (PM) by hydrolyzing an amide linkage between its aminonucleoside and O-methyl-L-tyrosine moieties {Nishimura et al . (1995) FEMS Microbiol . Lett . 132, 95-100} . In this study, we purified to homogeneity the enzyme from the cell-free extracts of S . morookaensis . The molecular weight of PM-hydrolyzing enzyme, estimated by SDS-PAGE and gel filtration, was 68 and 66 kDa, respectively, suggesting that this protein is monomeric . The PM-hydrolyzing activity was strongly inhibited by Zn2+, Fe2+, Cu2+, Hg2+, and N-bromosuccinimide, but was stimulated by DTT . The optimum pH and temperature for PM-hydrolyzing activity were 8.0 and 45 degrees C, respectively . Several L-aminoacyl-beta-naphthylamides were good substrates for the enzyme, suggesting that the PM-inactivating enzyme has an aminopeptidase activity . The N-terminal sequence of the first 14 amino acids (Val-Ser-Thr-Ala-Pro-Tyr-Gly-Ala-Trp-Gln-Ser-Pro-Ile-Asp) of the enzyme showed no significant homology with any published hydrolase sequences. Compend Contin Educ Dent, 1997 Sep, 18(9), 881 - 4, 886, 888 passim; quiz 894 Genetic risk for severe periodontal disease; Newman MG; The discovery of a genetic marker that is highly associated with increased risk for severe periodontitis is a major breakthrough in the clinical management of all dental patients . The marker is not diagnostic; rather, it is a prognostic test, and it is used to identify patients who are much more susceptible to plaque . Individuals who have the marker have a 6 to 19 times higher chance of getting severe periodontitis than those who do not have the marker . It is estimated that 30% of the US population will test positive . This new information supplements existing microbiology and immunology research . A limitation with the established view of the etiology of periodontal disease makes it difficult to explain to patients why they are more or less at risk based solely on their level of oral hygiene . It has also been difficult to predict the clinical outcomes of various treatments for individual patients because each person responds differently to their own plaque . The genetic discovery helps to explain why some people with a little plaque have a lot of disease, and why other people with a lot of plaque have only minor problems . By incorporating the constant factor, genetics, into your philosophy of practice, the patient's periodontal needs and desires can be better understood and managed. Med Tr Prom Ekol, 1998, (1), 13 - 6 {State of local immunity under exposure to anthropogenic factors of biological, chemical and physical nature in industry}; Litovskaia AV et al.; The authors studied immunologic features of saliva in 1714 workers exposed to vibration and other occupational hazards in microbiologic, chemical enterprises . The examinees demonstrated lower activity of lysozyme and concentrations of IgA, higher levels of IgG . Immunologic features of saliva was proved to have extreme diagnostic importance, therefore could be used to detect early signs of exposure to occupational hazards and to diagnose pathologic conditions caused by those hazards. Epidemiol Infect, 1998 Feb, 120(1), 37 - 42 Surveillance of outbreaks of waterborne infectious disease: categorizing levels of evidence; Tillett HE et al.; Public health surveillance requires the monitoring of waterborne disease, but sensitive and specific detection of relevant incidents is difficult . The Communicable Disease Surveillance Centre receives information from various sources about clusters of cases of illness in England and Wales . The reporter may suspect that water consumption or recreational water exposure is the route of infection, or subsequent investigation may raise the hypothesis that water is associated with illness . It is difficult to prove beyond reasonable doubt that such a hypothesis is correct . Water samples from the time of exposure are seldom available, some organisms are difficult to detect and almost everyone has some exposure to water . Therefore, we have developed a method of categorizing the degree of evidence used to implicate water . The categories take into account the epidemiology, microbiology and water quality information . Thus outbreaks are classified as being associated with water either 'strongly', 'probably' or 'possibly' . This system allows a broad database for monitoring possible effects of water and is not confined to the few outbreaks which have been intensively investigated or have positive environmental microbiology . Thus, for reported incidents, the sensitivity of classifying it as water associated should be high but this may be at the expense of specificity, especially with the 'possible' association. Rinsho Byori, 1998 Feb, 46(2), 124 - 31 {External quality assessment for clinical microbiology and good laboratory management}; Kumasaka K; The Tokyo Metropolitan external quality assessment (EQA) program has revealed some serious problems in private independent microbiology laboratories in Tokyo since 1982 . The poor performance in the EQA surveys closely related to poor laboratory managements, the type of training, experience of the medical technologists or technicians, and supervisory ability of the consultant physicians in independent laboratories . Social factors impede the reform of the quality assurance of clinical microbiology . Such factors include poor infrastructure of continuing education for small private laboratories, closure of the central clinical laboratories in the hospitals and outsourcing of laboratory tests due to restructuring in response to economic problems, and limited numbers of certified clinical pathologists of the Japan Society of Clinical Pathology (JSCP) . Therefore, the Tokyo Metropolitan EQA Scheme is still confidential and its main role is educational . Good two way communication between participants and the organizers' clinical pathologists is essential, if the quality of laboratory tests is to be improved . The new JSCP edition of the postgraduate training requirement in clinical pathology includes "Laboratory Administration and Management" . Good laboratory management(GLM) is an increasingly important component of good laboratory practice . The practice activities of clinical pathologists must include general management in addition to exercising there specialized knowledge in medicine and technology . Whereas leadership of a good clinical pathologist provides the direction of where a good laboratory is going, good management provides the steps of how to get there . And I believe quality system models from business and industry may provide us with strong guidance to build a quality system for the good laboratory that will endure into the next century. Rev Invest Clin, 1997 Sep-Oct, 49(5), 369 - 72 {Bibliometric repercussions of adopting English as the sole language for publication}; Bracho-Riquelme RL et al.; OBJECTIVE: To determine the effect that the change from French to English had on the impact of the Annales de l' Institut Pasteur . THEORETICAL REFERENCE: The Pasteur Institute journals have a prestige of over 100 years of existence and eight Nobel Prize winners in Physiology and Medicine . Changes through time on the impact of these journals can be monitored as they are included in databases of the ISI (Institute for Scientific Information) . METHODS: A year by year analysis from 1974 to 1992 was done using simple regression between percentage of articles published in English and: a) their impact factor; b) their ranking among journals of the same field . RESULTS: The determination coefficient (r2) between the percentage in English and the impact factor was 0.108, and that between percentage in English and the journal's rank in similar publications was 0.178 for the Ann Microbiol, 0.045 for the Ann Immunol and 0.122 for the Ann Virol . CONCLUSIONS: The change of language did not increase the impact factor of the French journals. J Periodontol, 1998 Feb, 69(2), 190 - 4 Microbiota associated with experimental peri-implantitis and periodontitis in adult Macaca mulatta monkeys; Eke PI et al.; This study examines the microbiota associated with the progression of experimental peri-implantitis and periodontitis induced concurrently in partially edentulous adult monkeys . Root-form and plate-form implants with fixed prosthesis in place for at least 12 months and their corresponding opposite molar teeth were ligated for 6 months . The microbiota in plaque around these ligated dental implants and molars were studied at 0, 1, 2, 3, and 6 months post-ligation . Plaque samples were analyzed by dark-field microscopy and selective and non-selective culture . Putative periodontal pathogens were detected as a major component of the microbiota cultured from plaque samples obtained from experimental peri-implantitis sites . Overall, the types and relative proportions of putative periodontal pathogens in plaque associated with ligature-induced peri-implantitis and ligature-induced periodontitis were similar . Only levels of anaerobic Actinomyces and spirochetes were significantly different between both sites . Spirochete levels were significantly higher at peri-implantitis sites when compared with levels at periodontitis sites after 6 months, and spirochete levels increased significantly between 0 and 6 months post-ligation at implant sites . Levels of spirochetes correlated significantly with probing depth and bone loss at peri-implantitis sites . Overall, Actinomyces levels were higher at periodontitis sites . Porphyromonas species were not detected continuously as part of the peri-implantitis microbiota . In conclusion, this study finds that the microbiota associated with the progression of experimental peri-implantitis and periodontitis occurring concurrently in partially edentulous mouths are similar. Int J Dermatol, 1998 Jan, 37(1), 1 - 6 Dermatoepidemiology . III . ABC principles for a critical review of the literature; Chuang TY et al.; EPIDEMIOLOGY: Is the study of disease occurrence in human populations . As a science, epidemiology emphasizes descriptive and analytic observation, clinical trial, behavioral intervention, and the practical utility of diagnostic tests . "Epidemiology" is derived from the Greek epi (among), demos (people), and logos (doctrine) . CLINICAL EPIDEMIOLOGY: Is the application of epidemiologic principles and methods to problems arising in clinical medicine, dermatology included . For dermatologists, understanding this discipline is as important as mastering other basic sciences, such as immunology, microbiology, and dermatopathology . The recognization of Lyme disease is a classic work of "infectious" disease epidemiology . In 1972, a disease characterized by erythema chronicum migrans and "endemic arthritis" clustered in Lyme, Connecticut . By 1975, an infectious agent was suspected to be the cause of the disease . In 1977, the tick was thought to be the vector; in 1980, the spirochete became the prime suspect and, in 1982, Borrelia burgdorferi was identified as the etiologic agent . The study of hexachlorobenzene exposure, resulting in porphyria turcica, is an example of classic "chronic" disease epidemiology . The illness began in 1955 when sporadic cases of porphyria occurred in eastern Turkey . In 1957, the first case with illness resembling congenital erythropoietic porphyria was described . In subsequent years, over 3000 patients developed "epidemic" porphyria . The cause was due to the ingestion of seed wheat which had been treated with fungicides containing 20% hexachlorobenzene . EPIDEMIOLOGIC METHODS: For research, published elsewhere as Dermatoepidemiology . I., include descriptive observational study, analytic observational study, epidemiologic experimental study and tests for sensitivity, specificity, and positive/negative predictive value . Epidemiologic principles, instead, stress the correct interpretation of data, minimization of bias, and the appreciation of natural variations in collected data. Clin Ther, 1998 Jan-Feb, 20(1), 88 - 100 Comparison of ceftibuten once daily and amoxicillin-clavulanate three times daily in the treatment of acute exacerbations of chronic bronchitis; McAdoo MA et al.; In medical practice, antibiotics are generally given empirically for the treatment of acute exacerbations of chronic bronchitis (AECB) . To be effective, antibiotic therapy should be broad in spectrum, and it should also cover the common beta-lactamase-producing pathogens . In this multicenter, randomized, investigator-masked study, 469 patients with AECB were randomized (in a ratio of 2:1) to receive 400-mg oral ceftibuten capsules once daily or 500-mg amoxicillin-clavulanate tablets three times daily for 5 to 15 days . Patients receiving ceftibuten were further divided into those who took the capsule with a meal (fed) and those who took the capsule 1 hour before a meal (fasted) . Clinical and microbiologic responses were evaluated after treatment at 0 to 6 days (end of treatment) and 7 to 21 days (follow-up) . Overall clinical success was determined by cure/improvement of signs and symptoms of AECB at the end of treatment and at follow-up . Overall microbiologic assessment was graded as eradication, persistence, relapse, reinfection, colonization, superinfection, or unassessable . Tolerability was evaluated by grading observed adverse events . The mean duration of treatment was 10.4 days for patients who received ceftibuten and 10.1 days for patients who received amoxicillin-clavulanate . A total of 252 patients receiving ceftibuten and 117 patients receiving amoxicillin-clavulanate were evaluable for clinical efficacy, and 55 patients were evaluable for microbiologic response . Both treatments improved the signs and symptoms of bronchitis, and overall clinical success rates were equivalent for patients treated with ceftibuten (211 of 252 {84%}) and amoxicillin-clavulanate (93 of 117 {79%}) (95% confidence interval {CI}, -4.5% to 13.6%) . Overall microbiologic eradication rates were also similar for patients treated with ceftibuten (36 of 37 {97%}) and amoxicillin-clavulanate (12 of 14 {86%}) (95% CI, -5.2% to 21.2%) . The most frequently reported treatment-related adverse events were gastrointestinal disturbances, which occurred in 15% (47 of 316) and 24% (36 of 152) of patients treated with ceftibuten and amoxicillin-clavulanate, respectively . No significant difference was observed in the ceftibutenfed and ceftibuten-fasted groups in overall clinical assessments of the clinical efficacy population and safety population . In conclusion, 400 mg oral ceftibuten once daily has a similar clinical success rate to 500 mg amoxicillin-clavulanate three times daily, with a trend toward fewer gastrointestinal side effects, in the treatment of patients with AECB. J Biol Chem, 1998 Feb 27, 273(9), 5155 - 66 Purification, regulation, and molecular and biochemical characterization of pyruvate carboxylase from Methanobacterium thermoautotrophicum strain deltaH; Mukhopadhyay B et al.; We discovered that Methanobacterium thermoautotrophicum strain DeltaH possessed pyruvate carboxylase (PYC), and this biotin prototroph required exogenously supplied biotin to exhibit detectable amounts of PYC activity . The enzyme was highly labile and was stabilized by 10% inositol in buffers to an extent that allowed purification to homogeneity and characterization . The purified enzyme was absolutely dependent on ATP, Mg2+ (or Mn2+ or Co2+), pyruvate, and bicarbonate for activity; phosphoenolpyruvate could not replace pyruvate, and acetyl-CoA was not required . The enzyme was inhibited by ADP and alpha-ketoglutarate but not by aspartate or glutamate . ATP was inhibitory at high concentrations . The enzyme, unlike other PYCs, exhibited nonlinear kinetics with respect to bicarbonate and was inhibited by excess Mg2+, Mn2+, or Co2+ . The 540-kDa enzyme of A4B4 composition contained a non-biotinylated 52-kDa subunit (PYCA) and a 75-kDa biotinylated subunit (PYCB) . The pycB gene was probably monocistronic and followed by a putative gene of a DNA-binding protein on the opposite strand . The pycA was about 727 kilobase pairs away from pycB on the chromosome and was probably co-transcribed with the biotin ligase gene (birA) . PYCA and PYCB showed substantial sequence identities (33-62%) to, respectively, the biotin carboxylase and biotin carboxyl carrier + carboxyltransferase domains or subunits of known biotin-dependent carboxylases/decarboxylases . We discovered that PYCB and probably the equivalent domains or subunits of all biotin-dependent carboxylases harbored the serine/threonine dehydratase types of pyridoxal-phosphate attachment site . Our results and the existence of an alternative oxaloacetate synthesizing enzyme phosphoenolpyruvate carboxylase in M . thermoautotrophicum strain DeltaH (Kenealy, W . R., and Zeikus, J . G . (1982) FEMS Microbiol . Lett . 14, 7-10) raise several questions for future investigations. Chest, 1998 Mar, 113(3 Suppl), 188S - 193S Nosocomial pneumonia guidelines: an international perspective; Mandell LA et al.; Hospital-acquired pneumonia is a serious illness with substantial morbidity and mortality . Management of this illness is challenging for the physician and a number of diverse issues must be considered when initiating therapy . Guidelines for the treatment of hospital-acquired pneumonia have been developed in Canada and the United States . A questionnaire sent to infectious disease physicians or clinical microbiologists in 29 countries showed that Australia, Sweden, and France had national guidelines in addition to Canada and the United States, while Hong Kong and France had single hospital-based guidelines . These guidelines are reviewed and some of the controversial issues relating to nosocomial pneumonia are discussed. J Gynecol Obstet Biol Reprod (Paris), 1997, 26(8), 781 - 8 {Diagnostic difficulties in infiltrating lobular breast cancer: value of ultrasonography . 33 cases}; Escolano E et al.; Thirty three invasive lobular carcinoma (ILC) were submitted to mammography, ultrasonography and finally surgery . The type of tumor proliferation and the absence of microcalcifications within the invasive tissue led to 15% of false negative responses in the mammographic analysis . Ultrasonography disclosed only 12% of false negatives . Sonographic appearance, especially fine needle aspirations or microbiopsy under ultrasonographic control, allows modification of mammographic and clinical diagnosis errors . Thus, this method appears of importance in diagnosis of ILC. Eur J Cardiothorac Surg, 1998 Jan, 13(1), 71 - 7 Factors affecting the yield of cardiac valve allografts from living unrelated donors; Hunt CJ et al.; OBJECTIVE: Allografts are the valve of choice for fertile women, patients with infective endocarditis and those with small aortic roots . However, the supply of valves is problematic and widespread usage is restricted by limited availability . Allograft valves are available from cadaveric donors and from the explanted hearts of transplant recipients . Potentially, hearts from these patients could be an excellent source of usable aortic and pulmonary valves . However, little information is available on the suitability of such donors, the procurement rate of allograft valves from this source, or the factors that limit the yield of implantable valves from explanted hearts . METHOD: In order to examine some of these issues, we have carried out a retrospective study on the explanted hearts offered to the East Anglian Tissue Bank by Papworth hospital . Papworth hospital carries out approximately 90 heart and heart/lung transplants per year . Over a 2 year period, the tissue bank was offered 72 hearts from this programme . RESULTS: Of the 72 hearts offered, 58 were accepted for subsequent dissection and further examination . A total of 14 hearts were refused . The main reasons for refusal were extensive cardiectomy trauma (4 hearts) and abnormal valve morphology (four hearts) . Of the 116 valves from those hearts accepted for dissection, 55 valves were rejected upon further examination . Reasons for rejection included: cardiectomy trauma (26 valves), abnormal morphology (22 valves), procurement/dissection trauma (7 valves) . Of the 61 valves banked, four were subsequently rejected due to positive or incomplete microbiology . Procurement trauma fell to 0% in the last 12 months of the study but cardiectomy trauma remained constant and was related to previous cardiac surgery . Overall, the yield of implantable valves was 0.8 valves/donor . However, the yield showed considerable variation, from 1.0 valves/donor for donors diagnosed as cardiomyopathy to 0.5 valves/donor for donors with ischaemic heart disease who had undergone previous cardiac surgery . CONCLUSION: It is possible to predict the likely yield of explanted heart valves from different groups of heart transplant recipients, based on diagnosis and previous history . The yield of usable valves could be increased by avoidance of injury, both during cardiectomy and subsequent removal of the valves; this is achievable through appropriate training. Gac Med Mex, 1997, 133 Suppl 1, 111 - 24 {Advances in the molecular diagnosis of infectious and parasitic diseases}; Sweet-Cordero A et al.; During the last decade molecular diagnostic techniques have moved from the research laboratory into the clinical microbiology laboratory . The application of molecular biology for the diagnosis of infections and parasitic diseases by the detection of nucleic acids has steadily grown, and it is very probably that, while they may not displace the traditional diagnostic laboratory, they will be common place in the not to distant future . A detailed description of the principal molecular diagnostic techniques that are currently being used or that have a potential use for the diagnosis, evaluation of disease progression or response to therapy of selected infectious and parasitic diseases, is presented . Emphasis is placed on the rational use of these techniques in regional reference laboratories or highly specialized hospitals; the importance of selecting and implanting those diagnostic techniques with the highest cost-benefit ratio; and finally, the need to train human resources which are highly qualified in the theory and practice of molecular biology. Allerg Immunol (Paris), 1998 Jan, 30(1), 18 - 24 {Updates on the earlier treatments for atopic dermatitis}; Jelen G; The GERDA classes have the function of updating our knowledge of dermato-allergology . One of the themes tackled this year was the treatment of atopic dermatitis . Apart from consideration of treatment or exception with cortisone, it seemed to be of interest to find the relevance of "old treatments" for atopic dermatitis, either preventive or symptomatic . Preventive treatment made reference to correction of food factors (diet in infants, removal of maternal allergens, supplementation on fatty acids) and of environmental factors especially the fight against house dust mites by use of anti-mite mattress covers . Miracle treatments of atopy do not always exist . Thus there is often need for, besides local corticosteroid therapy, an external symptomatic treatment where the emphasis is on the struggle against skin microbiology, the fight against pruritic inflammatory conditions and above all the battle against xerosis . Knowledge of the physiology of the stratum corneum gives better understanding of the effect of emollients and moisturizers in restoration of the cutaneous barrier, of which dysfunction is one of the elements of atopic dermatitis. J Radiol, 1997 Nov, 78(11), 1141 - 6 {Stereotactic core biopsy of breast microcalcifications . Aid to diagnosis?}; Chopier J et al.; We assessed the accuracy with which stereotactic core biopsy of breast microcalcifications helps to justify follow-up or indicate required surgical planning . Eighty-nine patients underwent stereotactic breast biopsy performed with large core needles . Histopathologic findings of microbiopsies were correlated with radiological findings, with follow up and with surgical findings . Agreement between radiological and histopathologic findings on biopsies was obtained in 86% for benign results . Even in the case of benign radiological findings, the histological results of microbiopsies required a surgical biopsy justified in 10% of cases . Radiologically suspicious or malignant microcalcifications were correlated with 35% benign and 65% malignant histologies on biopsies . The positive predictive value, negative predictive value and accuracy were respectively 93%, 86% and 95% . Invasive lesions were found on microbiopsies in 39% of excised malignant lesions . Multifocal lesions were proved on microbiopsies in 50% of all excised carcinomas . Microbiopsies help to disclose malignant lesions undetected on radiological analysis, and may obviate diagnostic surgical biopsy in many cases. Virology, 1998 Mar 1, 242(1), 68 - 79 An equine herpesvirus type 1 recombinant with a deletion in the gE and gI genes is avirulent in young horses; Matsumura T et al.; The cell culture-adapted KyA strain of equine herpesvirus type 1 (EHV-1) has been found to be attenuated in young horses (Matsumura et al., 1996, Vet . Microbiol . 48, 353-365) . The KyA strain lacks at least six genes in its genome, including those encoding glycoproteins gE and gI . To elucidate whether EHV-1 glycoproteins gE and gI play a role in viral virulence, we have constructed an EHV-1 recombinant that has the genes encoding both gE and gI deleted from its genome and its revertant . Growth properties of the deletion mutant virus in vitro were compared with those of the parent and the revertant viruses . Plaque size of the mutant virus in fetal horse kidney (FHK) cells was significantly smaller than those of the parent and the revertant viruses . In one-step growth experiments, however, the yields of infectious virus from FHK cells infected with the deletion mutant, the parent, or the revertant virus were approximately the same . The results suggested that gE and/or gI of EHV-1 promoted cell-to-cell spread of the virus, but that these glycoproteins were not involved in the process of virus maturation and release or in virus attachment and penetration . Subsequently, the virulence of mutant and revertant viruses was examined in young horses . No clinical signs were observed in six horses, including three colostrum-deprived foals inoculated intranasally with the deletion mutant virus, whereas three colostrum-deprived foals inoculated intranasally with the revertant virus manifested clinical signs typical for EHV-1 respiratory infection (i.e., pyrexia, nasal discharge, and swelling of submandibular lymph nodes) . The results obtained from in vivo studies revealed that the EHV-1 mutant defective in both gE and gI genes was avirulent in young horses, suggesting that gE and/or gI of the EHV-1 have an important role in EHV-1 virulence . However, the EHV-1 mutant defective in both gE and gI genes induced only a partial protectivity in inoculated foals from manifestation of respiratory symptoms after challenge infection. Int J Periodontics Restorative Dent, 1997 Aug, 17(4), 334 - 47 Follow-up of two cases of Papillon-Lefèvre syndrome and presentation of two new cases; Boutsi EA et al.; Periodontitis resulting from Papillon-Lefevre Syndrome has been known to cause early loss of primary dentition with subsequent involvement of the permanent dentition . In this study, two Papillon-Lefevre Syndrome patients were followed for 3 years after initial treatment and improvement of their periodontal condition . In addition, two new cases of Papillon-Lefevre Syndrome are presented . The follow-up treatment of the first two patients included monitoring the oral hygiene and performing ultrasonic scaling . Their present clinical appearance is very satisfactory . The periodontal condition of the third (new) patient was brought under control by extracting the involved deciduous teeth under antibiotic coverage, and by scaling and root planing the already erupted permanent teeth as well as by maintaining a high standard of oral hygiene . In the fourth case, all permanent teeth had erupted and the periodontium had already been severely destroyed . Actinobacillus actinomycetemcomitans was not detected by microbiologic examination after the periodontal conditions improved, except in the fourth case . Western blot analysis showed that the three first three patients had positive antibody response to the same antigens of Actinobacillus actinomycetemcomitans . Phagocytosis by polymorphonuclear neutrophil leukocytes) had not decreased, but the expression of surface receptors of polymorphonuclear neutrophil leukocytes was within the normal limits. Bone Marrow Transplant, 1998 Jan, 21(2), 195 - 203 A detailed analysis of charges for hematopoietic stem cell transplantation at a children's hospital; Kline RM et al.; We analyzed hospital charges for pediatric hematopoietic stem cell transplantation (HSCT) to understand better the medical origin of these charges . Forty-nine patients undergoing HSCT at Kosair Children's Hospital between January 1992 and August 1995 had hospital charges analyzed by cost center, donor type and clinical outcome . Thirty-three autologous, two syngeneic and 14 allogeneic transplants were performed . Twenty-four transplants were performed for hematological malignancies, 22 for solid tumors, and three for non-malignant diseases . Pharmaceutical charges comprised the largest single component of total hospital charges (THC), accounting for 38.9% . Room charges were the next largest group at 33.7% of THC . Other cost centers, in order of magnitude, were central supply (7.9%), transfusion services (7.5%), laboratory (5.8%), microbiology (3.6%), miscellaneous (1.9%), and radiology (1.4%) . Within the pharmaceutical cost center, colony-stimulating factors comprised the largest single item, making up 18% of total pharmacy charges and 7% of THC . Antibiotics were the second largest component, at 16% of pharmacy charges and 6% of THC . Patients transferred to the intensive care unit (ICU) had charges 68% greater than non-ICU patients . Allogeneic transplant patients had THC 35% greater than autologous transplant patients, but also a four-fold greater chance of becoming an ICU patient . THC for non-ICU allogeneic transplant patients were 18% greater than for autologous non-ICU patients . THC for allogeneic ICU patients were 21% greater than for autologous ICU patients . Patients who died of transplant-related toxicity prior to day 100 had THC 83% greater than those who survived beyond day 100 . This is the first published comprehensive and detailed analysis of charges associated with hematopoietic stem cell transplantation . With increased emphasis on the provision of cost-effective care in both Europe and the USA, medical practices must be examined with the goal of reducing inefficiencies while preserving quality of care . Understanding the genesis of charges in expensive procedures such as stem cell transplantation is an initial step in cost containment. Bratisl Lek Listy, 1997 Oct, 98(10), 559 - 62 {Epidemiologic and microbiologic aspects of mycobacteriosis in Slovakia . M . fortuitum}; Badalik L et al.; The purpose of this paper is to confront the knowledge about the epidemiology of this disease abroad with information obtained in Slovakia . The methodology of this paper is based upon surveillance of tuberculosis as applied on mycobacterioses . Pulmonary infections caused by M . fortuitum presents only 10% of pulmonary mycobacterioses caused by facultative pathogenic mycobacteria . Since the year 1979 were in the territory of Slovakia newly discovered 5 cases of mycobacterioses caused by M . fortuitum . 4 of them are men (age 61-77 years old) and 1 women (57 years old) . 3 patients are from Eastern Slovakia and 2 patients are from Central Slovakia . All of them had pulmonary location of disease . Another 8 cases (5 men and 3 women) are suspect for mycobacterioses for the meantime . In each isolation of the agents was made 5-8 times but without verification of clinical activity . Findings of M . fortuitum in biological material even when repeated can mean contamination only . During the period of last 10 years 768 isolations of M . fortuitum from 496 persons were registered . (Ref . 36.) Pathologica, 1997 Oct, 89(5), 495 - 502 {Origin and development of pathological anatomy in Trieste: a paradigmatic history}; Melato M; The history of the Italian city of Trieste is somewhat unique . In 1382, being unable to compete with nearby Venice, it gave itself up to the Archduchy of Austria, which, while ensuring Trieste a certain degree of autonomy, did nothing to improve the town's economy . Things changed, however, after 1719, when Trieste was declared a Free Port . The city, at the time poor and lacking the human and economic resources that might have given life to the long-awaited trading activities, started to attract flocks of immigrant labourers and professionals, including many physicians . Thus, from mid-18th C onwards, Trieste witnessed the rise of a local class of physicians . Graduates from the major European universities of the times, these physicians were highly active in the rapid professional and cultural growth of the city . Being free from traditions or affiliations to specific medical schools, this unusually multicultural class of professionals provides a sample of the times' medical class which is particularly suitable for understanding what schools of thought dominated during that period . A number of publications, largely dealing with pathology, and many historical circumstances show that this newly formed class of physicians was influenced by the scientific method advanced by Morgagni in his De Sedibus . It is also clear that Morgagni's influence greatly benefited the city's doctors and soon turned them into fertile and original promoters of ideas, as well as highly skilled professionals . When the times became ripe to take on a specialist in pathology, both medical class and local authorities showed great interest in the field by making sure that the successful candidate, Dr . Simon Pertot, had all the resources and equipment necessary for the specialist to become a central figure in professional training and medical research in Trieste . Thus, the kind of clinical pathology where a single physician performed macroscopic examinations to identify his patient's cause of death gave way to a more specialised discipline which was able to transfer--to post-mortem examinations first and soon after to surgically removed tissues--the wealth of notions developed in the middle of the 19th C in the basic sciences of chemistry, microbiology and physiology, as had been advanced by the second founding father of the discipline, Virchow . Trieste, therefore, may be said to offer a unique example of the origins and development of pathologic anatomy in Central Europe . And the recollection of the city's experience promotes, in the modern pathologist, an awareness of belonging to a discipline which has always been the soundest leader in the advancement of medicine. Can J Cardiol, 1998 Jan, 14(1), 85 - 6 Purulent pericarditis secondary to Candida parapsilosis and Peptostreptococcus species; McNamee CJ et al.; Purulent pericarditis secondary to Candida pericarditis is a rare but lethal infection . The first case of pericardial infection by Candida parapsilosis in combination with Peptostreptococcus species in an immunocompetent nondebilitated individual is reported . Urgent pericardiectomy was required after pericardial drainage failed to stop symptoms of tamponade . The etiology, microbiology and treatment of this unusual condition are reviewed. Tidsskr Nor Laegeforen, 1998 Jan 10, 118(1), 67 - 70 {A classification system of anogenital findings . A diagnostic tool in the assessment of sexually abused children}; Myhre AK et al.; The assessment of suspected sexual child abuse demands teamwork where the paediatrician plays a central role . From a juridical point of view, the task of the paediatrician is to evaluate the anatomic, microbiologic and forensic medical findings . In 1995, in order to improve the quality of this work, Norwegian paediatricians established a peer review group which meets on a regular basis . Based on available literature and the experience of the individual members, a classification system for anogenital findings has been developed . The findings are divided into five classes . Class one comprises findings frequently seen in children who have not been abused . Class two comprises findings not considered to be normal, but for which there could be many different causes . Classes three, four and five represent findings which are increasingly predictive with respect to injury penetration or attempted penetration . Since our knowledge of anogenital anatomy in children who have not been abused is limited, our classification system should be updated regularly. Histol Histopathol, 1998 Jan, 13(1), 129 - 36 Characterization of natural occurring Pneumocystis carinii pneumonia in pigs by histopathology, electron microscopy, in situ hybridization and PCR amplification; Ramos Vara JA et al.; Macroscopic, histologic, ultrastructural, microbiologic, in situ hybridization (ISH) and PCR detection results in three 8-week-old pigs naturally infected with Pneumocystis carinii (PC) are described . All animals had a nonsuppurative interstitial pneumonia and intra-alveolar Pneumocystis organisms with foamy eosinophilic and PAS positive appearance . Ultrastructurally, PC trophozoites and cysts were observed in pigs No . 2 and No . 3, with the former being much more numerous . PC organisms were located on the alveolar surface or within the alveolar septa . Trophozoites had numerous filopodia and were thick-walled . Cysts had no or few filopodia, were thick-walled and contained intracystic bodies . Using non-isotopic ISH on formalin-fixed, paraffin-embedded lung tissue sections, PC DNA from pigs No . 2 and No . 3 hybridized with a probe specific for PC ribosomal RNA (rRNA) . Using primers specific for mitochondrial rRNA gene (pAZ102-E/pAZ102-H), and for the internal transcriber spacers of ribosomal gene of PC, PCR methods amplified a product in the lung of pigs No . 2 and No . 3 using either frozen or formalin-fixed and paraffin-embedded lung tissue . DNA from Pig No . 1 samples did not amplify with any primer . This is the first time that molecular biology techniques (in situ hybridization and PCR) have been applied to the study of porcine pneumocystosis. CLAO J, 1998 Jan, 24(1), 48 - 51 Twice-a-day versus four-times-a-day ofloxacin treatment of external ocular infection; Friedlaender MH; PURPOSE: The purpose of this study was to compare the efficacies of 0.3% ofloxacin eyedrops, when given twice-a-day (BID) versus four-times-a-day (QID), for the treatment of external ocular disease . METHOD: Fifty patients with blepharitis, conjuctivitis, or blepharoconjunctivitis were randomly assigned to treatment with 0.3% ofloxacin eyedrops, BID or QID, for 10 days . Signs, symptoms, and cultures were evaluated at the beginning and at the end of the study . RESULTS: The clinical outcome was virtually identical in the two groups . There was a significant decrease in clinical scores in the BID and QID groups by days 3 to 5 (2.6-3.0 points) and a further decrease by day 11 (4.3-5.0 points) . There was no significant difference between the two groups at any time interval . Microbiologic studies showed a reduction in colony-forming units in 87% of the BID group and in 80% of the QID group . CONCLUSION: The treatment of external ocular disease with 0.3% ofloxacin eyedrops was equally effective when given BID or QID. BMJ, 1998 Jan 24, 316(7127), 253 - 8 Epidemiology and detection of HIV-1 among pregnant women in the United Kingdom: results from national surveillance 1988-96; Nicoll A et al.; OBJECTIVE: To describe the epidemiology of HIV-1 infection in pregnant women in the United Kingdom . DESIGN: Serial unlinked serosurveillance for HIV-1 in neonatal specimens and surveillance through registers of diagnosed maternal and paediatric infections from reporting by obstetricians, paediatricians, and microbiologists . SETTING: United Kingdom, 1988-96 . SUBJECTS: Pregnant women proceeding to live births and their children . MAIN OUTCOME MEASURES: Time trends in prevalence of HIV-1 seropositivity in newborn infants (as a proxy for infection in mothers); the proportions of mothers with diagnosed HIV-1 infections, and their characteristics . RESULTS: HIV-1 prevalence among mothers in London rose sixfold between 1988 and 1996 (0.19% of women tested; 1 in 520 in 1996) . Apart from in Edinburgh and Dundee, levels remained low in Scotland (0.025%; 1 in 3970) and elsewhere in the United Kingdom (0.016%; 1 in 1930) . Over a third of births to infected mothers in 1996 occurred outside London . In London the reported infections were predominantly among black African women, whereas in Scotland most were associated with drug injecting . The contribution of reported infection among African women increased over time as that of drug injecting declined . In Scotland 51% of mothers' infections were diagnosed before the birth . In England, despite a national policy initiative in 1992 to increase the antenatal detection rate of HIV, no improvement in detection was observed, and in 1996 only 15% of previously unrecognised HIV infections were diagnosed during pregnancy . CONCLUSIONS: HIV-1 infection affects mothers throughout the United Kingdom but is most common in London . Levels of diagnosis in pregnant women have not improved . Surveillance data can monitor effectively the impact of initiatives to reduce preventable HIV-1 infections in children. Epidemiol Mikrobiol Imunol, 1997 Sep, 46(3), 124 - 7 {Today's tasks in medical mycology}; Jesenska Z; The author analyzes contemporary tasks of mycology . Problems comprise clinical as well as environmental mycology which possesses a large amount of new findings and thus offers many research opportunities for young microbiologists. Acta Microbiol Immunol Hung, 1997, 44(3), 291 - 4 The development and results of oral microbiology at the Semmelweis University of Medicine; Banoczy J et al.; The development of oral microbiology at the Semmelweis University of Medicine started around 1980 in the Institute of Microbiology . The teaching of oral microbiology was introduced to the dental educational program, basic research was initiated, appropriate methods learned and used . An intensive collaboration with the Department of Conservative Dentistry developed, which resulted between 1983 and 1995 in about 23 publications (the majority in international journals) and eight lectures, mainly on international forums. Oral Microbiol Immunol, 1996 Dec, 11(6), 425 - 7 Distribution of a newly described species, Kingella oralis, in the human oral cavity; Chen C; The oral distribution of Kingella oralis was investigated in 10 periodontally healthy subjects . 11 untreated adult periodontitis patients and 6 untreated localized juvenile periodontitis patients . From each subject, 6-8 each of supra- and subgingival tooth samples, 4 mucosa samples and a saliva sample were examined by culture for the presence of K . oralis . K . oralis was found in at least one oral site in 26 of the 27 study subjects, and in at least one tooth site in each of these 26 positive subjects . Its prevalence in dental plaque ranged from 23% to 59% in different subject groups . The mean percentage of K . oralis in total microbiota in the dental plaque ranged from 0.40% in the periodontally healthy group to 4.60% in localized juvenile periodontitis subjects . The organism was a significant species in a few periodontitis sites, constituting > 5% of the total microbiota. Onderstepoort J Vet Res, 1997 Sep, 64(3), 195 - 9 Strains of Actinobacillus spp . from diseases of animals and ostriches in Zimbabwe; Mohan K et al.; Among the Actinobacillus spp . only A . lignieresii represents a homogenous and well studied taxon . However, haemolytic and non-haemolytic strains of A . equuli and A . suis are also isolated from a wide range of diseases in a variety of hosts . These isolates often pose problems in definitive identification . Consequently, several studies have been published, emphasizing the need for detailed studies to reclassify various members of this genus and also to assess their disease significance . We isolated 48 strains of Actinobacillus from clinical cases in horses, cattle, sheep, cat, pigs and ostrich . In order to investigate the association of various taxa of Actinobacillus in different pathological conditions in these hosts, the Zimbabwean isolates were characterized in detail and assigned to the recently described taxa of Actinobacillus . Representative strains of different taxa were also confirmed at the Department of Veterinary Microbiology, Frederiksburg C, Denmark . Of the 48 isolates, 22 were identified as A . lignieresii, 13 as A . equuli; 6 as A . suis, four as belonging to Taxon 11 and one as belonging to Taxon 9 . These two taxa were recently described . Details of pathological conditions in the respective hosts and the significance of isolation, are discussed . We have reported for the first time isolation of A . lignieresii from a post-operative wound in a cat and there is also a first report of isolation from an ostrich . We considered A . equuli to be the primary cause of equine abortion and septicaemia, and Taxon 9 as causing "sleepy foal" disease . We did not encounter any case of "wooden tongue", but isolated A . lignieresii from cases of superficial lymphadenitis in cattle and sheep . This appears to be a first report of detailed descriptions of Zimbabwean strains of Actinobacillus spp. J Clin Microbiol, 1998 Feb, 36(2), 526 - 30 Evaluation of PCR primers for early diagnosis of cytomegalovirus infection following liver transplantation; Mendez JC et al.; The availability of microbiologic methods that detect early replication of cytomegalovirus (CMV) posttransplantation will enhance the process of initiating preemptive antiviral therapy prior to the appearance of CMV disease . Using PCR techniques we sought to determine which region of the CMV genome present in peripheral blood leukocytes (PBLs) or serum provides the highest sensitivity for the detection of CMV posttransplantation . Blood samples were prospectively collected weekly for at least 8 weeks from a cohort of 21 consecutive liver transplant recipients not receiving anti-CMV prophylaxis . Results of PCR assays were correlated with recovery of CMV in cell cultures and histopathological findings from biopsy specimens of infected organs to assess clinical symptomatic infection . Of 148 specimens, primer pairs directed to the HindIII-X fragment region of CMV detected target DNA with a 94% sensitivity, compared to an 87% sensitivity with primer pairs directed to EcoRI fragment D, 32% sensitivity with primer pairs directed to the immediate-early antigen 1 gene (IEA1 gene), and 20% sensitivity with primer pairs directed to the major immediate-early (MIE) gene . The performance characteristics in terms of the sensitivity of primers for amplifying CMV DNA associated with symptomatic infection ranged from 100% (HindIII-X) to 20% (MIE gene); however, specificity was inversely related (HindIII-X, 45%; MIE gene, 91%) to primers directed to these gene targets . When HindIII-X and EcoRI-D primer sets were used, CMV DNA from PBLs was a more sensitive target than CMV DNA from serum for the early detection of symptomatic CMV infection (17 versus 12 days) . Importantly, CMV DNA was not detected in five patients with no evidence of this viral infection . In conclusion, primers directed to the HindIII-X fragment region were the most optimal for the early detection of CMV DNA in PBLs and sera from symptomatic liver transplant recipients. Appl Environ Microbiol, 1998 Feb, 64(2), 492 - 5 Engineering of a cold-adapted protease by sequential random mutagenesis and a screening system; Taguchi S et al.; A cold-adapted protease subtilisin was successfully isolated by evolutionary engineering based on sequential in vitro random mutagenesis and an improved method of screening (H . Kano, S . Taguchi, and H . Momose, Appl . Microbiol . Biotechnol . 47:46-51, 1997) . The mutant subtilisin, termed m-63, exhibited a catalytic efficiency (expressed as the kcat/Km value) 100% higher than that of the wild type at 10 degrees C when N-succinyl-L-Ala-L-Ala-L-Pro-L-Phe-p-nitroanilide was used as a synthetic substrate . This cold adaptation was achieved with three mutations, Val to Ile at position 72 (V72I), Ala to Thr at position 92 (A92T), and Gly to Asp at position 131 (G131D), and it was found that an increase in substrate affinity (i.e., a decreased Km value) was mostly responsible for the increased activity . Analysis of kinetic parameters revealed that the V72I mutation contributed negatively to the activity but that the other two