Prevalence of gram-negative rods in the normal pharyngeal flora

We obtained throat cultures from 100 randomly selected people free from any chronic upper or lower respiratory disease who did not work in a hospital and who had not experienced any acute illness or received any antibacterial therapy in the 4 weeks preceding culture. Eighteen percent harbored either a species of Enterobacteriaceae or Pseudomonas aeruginosa in their pharynx. In all cases, colony counts were low, the majority being detected in broth media selective for Gram-negative rods. There were no clear-cut age or sex distributions of Gram-negative pharyngeal carriage. These data imply that, in at least some cases, isolation of Gram-negative rods from sputum of untreated patients may be a normal finding, and that in some patients with pulmonary infection, the pretreatment, upper respiratory tract flora may serve as the source of subsequent superinfection with Gram-negative rods.     

Occurrence of extended spectrum beta-lactamases (ESBL) and inducible beta-lactamases (IBL) in clinical strains of gram-negative rods

Hairy cell leukemia (HCL) is clinically associated with severe T-cell dysfunction. Several new observations have given more insight into the abnormal T-cell responses seen in this disease. T-lymphocytes in the spleen of patients with HCL seem to be abnormally activated. On the other hand, they are non-responsive, possibly as a result of monocytopenia which may lead to inadequate antigen presentation. This, together with the lack of CD28 on T-cells, may cause T-cell dysfunction. Furthermore, there is a very restricted repertoire of the T-cell receptor-beta family, which may also result in non-responsiveness. Otherwise, T-cell clonal excess may be indicative for activated, possibly autoreactive T-cells.     

Occurrence of gram-negative non-fermenting rods in hemocultures and their sensitivity to antimicrobial agents

In the period from January 1993 to June 1996 were at the Department of Microbiology of the University Hospital in Olomouc 122 strains of Gram-negative nonfermentative rod-shaped bacteria isolated from haemocultures. The majority represented the group of 51 strains of the genus Acinetobacter (41.8%), complex A. calcoaceticus-baumannii (Acb complex). The second largest group were 21 strains (17.2%) of Pseudomonas aeruginosa. These were followed by 17 strains (13.9%) of Stenotrophomonas maltophilia, 8 strains (6.6%) of non-Acb complex acinetobacters, 6 strains (4.9%) of Pseudomonas putida and 5 strains (4.1%) of Alcaligenes xylosoxidans. The remaining species were represented only by 1-2 strains. In three isolations was the identification impossible. The majority of strains (24.6%) were from the Department of Haematology of the University Hospital in Olomouc. The most frequent diagnoses in patients with positive haemocultures were leukemias and lymphomas (24.6%). The most effective tested antimicrobial agents were ceftazidime (93.4% of sensitive strains) and ofloxacin (91.7%). From the total number of 80 strains detected using the equipment BacT/Alert 120, 22 (27.5%) were isolated repeatedly confirming their role in the etiology of bacteriemic or septic episodes. Because only one blood sample was obtained in 34 cases (58.6%) of the remaining 58 only once detected strains, it was impossible to confirm their etiologic role by repeated isolation. (Tab. 6, Ref. 22.)     

Rapid detection of Shiga-like toxin gene in feces with rapid-cycle polymerase chain reaction

A rapid detection for Shiga-like toxin in feces was developed with the nucleic acid extraction method by silicondioxide-guanidine thiothianate and rapid-cycle polymerase chain reaction by RapidCycler (model 1002; Idaho Technology, RC-PCR here after). Twenty-two fecal samples that were collected from patients with diarrhoea caused by E. coli O157:H7 and frozen for 6 months were examined directly by RC-PCR, conventional PCR assay using by ThermalCycler 9600-R (Roche, TC-PCR here after) and by the culture method using tellurite-cefixime sorbitol MacConkey (direct method). These examinations were done also after being injected into TCV-TSB and incutated at 35 degrees C overnight (indirect method). The sensitivity of RC-PCR and TC-PCR using a diluted suspension of broth enriched at 35 degrees C overnight were 4.1 pg and 410 fg, respectively. Positive results in the direct method were obtained in 7 for RC-PCR, 10 for TC-PCR and 5 for culture. Positive results on indirect assay were obtained in 9 for RC-PCR, 9 for TC-PCR and 7 for culture. It was demonstrated that the RC-PCR assay was able to detect Shiga-like toxin gene in feces in less than 90 minutes after being received at the laboratory.     

Rapid diagnosis of asymptomatic hereditary haemochromatosis by detection of the Cys282Tyr mutation in the HLA-H gene

Hereditary haemochromatosis is an autosomal recessive disorder characterised by life-long excessive accumulation of iron. A candidate gene for hereditary haemochromatosis has recently been reported (HLA-H) and a specific missense mutation (Cys282Tyr) has been identified in 85% of patients with the disorder. We describe the rapid detection of this mutation using the polymerase chain reaction and restriction endonuclease digestion. The usefulness of this test for early diagnosis of hereditary haemochromatosis in asymptomatic family members is highlighted.     

Functional characteristics of lateral interactions between rods in the retina of the snapping turtle

1. Intracellular recordings were made of the slow hyperpolarizing light responses of single rods in the retina of the snapping turtle. Physiological criteria used to identify rods were verified by intracellular injections of Procion Yellow. 2. The amplitudes of the responses elicited by fixed intensity flashes increased as the stimulus was enlarged to a diameter of 300 mum. Scattered light was found incapable of accounting for this effect, which must result from summative interaction of rods with neighbouring receptors. Effects of summative interaction were observed even at stimulus intensities that produced maximal responses. Enlarging the diameter of the higher intensity stimuli from 100 to 300 mum increased the peak response amplitude by almost 50%; it also produced a distinct initial peak of the response which we term overshoot. The amplitude of this overshoot was graded with stimulus size. 3. Complete intensity-response relationships were determined using stimulus diameters of 100 and 750 mum for each rod. With the smaller stimulus the intensity response range was 4-5 log units, and with the larger stimulus this was increased to 5-0 log units. For intensities below about 60 quanta/mum2 per flash (514 nm) the amplitudes elicited by the large stimulus followed a sigmoid-shaped curve. However, at higher intensities an additional lobe appeared on the intensity-response relationship. The appearance of this lobe correlated with the emergence of the overshoot on the response wave form. 4. Determinations of rod flash sensitivity (mV per quantum per mum2) showed that it increased with stimulus size up to a stimulus diameter of about 300 mum. With diameters between 50 and 150 mum, a linear relationship existed between the flash sensitivity and stimulus area. Absolute quantal sensitivities increased with stimulus area by a factor of 26, from a value of 28 muV per photoisomerization per rod with a stimulus 25 mum in diameter, to 720 muV per photoisomerization per rod with a stimulus 300 mum in diameter. 5. By comparison, red-sensitive cones showed increased sensitivity as a function of stimulus size only up to a stimulus diameter of 120 mum. Their over-all sensitivity was lower than that of rods and proved linear with stimulus diameter rather than with stimulus area. 6. Simultaneous recordings were made from rod-cone pairs to determine whether the overshoot, and hence the lobe on the amplitude-intensity function, could result from a cone input to the rod response. The time course of the cone response proved much too rapid to fit the overshoot of the rod response. 7. The spectral sensitivity of the dark-adapted rod response closely followed the difference spectrum of the rod photopigment for wave-lengths greater than 450 nm. This was true throughout the intensity range of the response, including low intensities where response averaging was necessary. 8. At low response amplitudes (approximately 1 mV), about 70% of the 40 rods tested showed responses to long wave-length stimuli consisting of two components...     

Occurrence of homologs of the Escherichia coli lytB gene in gram-negative bacterial species

The Escherichia coli LytB protein regulates the activity of guanosine 3',5'-bispyrophosphate synthetase I (RelA). A Southern blot analysis of chromosomal DNA with the E. coli lytB gene as a probe revealed the presence of lytB homologs in all of the gram-negative bacterial species examined but not in gram-positive species. The lytB homologs from Enterobacter aerogenes and Pseudomonas fluorescens complemented the E. coli lytB44 mutant allele.     

Rapid detection of K-ras gene mutations in canine lung cancer using single-strand conformational polymorphism analysis

A total of 126 spontaneous lung tumors from pet dogs were examined for K-ras mutations within exon 1 and exon 2 using a non-radioisotope single-strand conformational polymorphism analysis (SSCP) detection method on PCR products. Mutations were confirmed by direct DNA sequencing. Tumors were classified as adenomas (9), bronchioloalveolar carcinomas (59), adenocarcinomas (30), adenosquamous carcinomas (16), squamous cell carcinomas (3) and anaplastic carcinomas (9). Nineteen mutations were detected in the malignant tumors: 18 occurred in exon 1 codon 12 and one in exon 2 codon 61. No mutations were present in the adenomas. The most common mutation was a G-->A transition (11/19) in the second position of codon 12. Based on this study, K-ras mutations occur in canine non-small cell lung carcinomas. The frequency and type of mutation more closely matches tumors from human non-smokers with K-ras mutations than smokers. With the application of screening techniques such as SSCP, large numbers of dog tumors can be examined to provide a large animal model for comparative studies of carcinogenesis.     

Antimicrobial activity of fluoroquinolones and other antibiotics on 1,116 clinical gram-positive and gram-negative isolates

A total of 1,116 clinically isolated strains belonging to Staphylococcus aureus (200), Staphylococcus epidermidis (200), Streptococcus pneumoniae (20), Escherchia coli (200), Klebsiella spp. (177), Serratia marcescens (22), Pseudomonas aeruginosa (224), Haemophilus influenzae (35) and Salmonella (38) from the Department of Infectious Diseases, La Sapienza University in Rome (Italy) were tested against three fluoroquinolones (ofloxacin, ciprofloxacin and levofloxacin) and 10 other antibiotics (augmentin, ampicillin, cefaclor, cefixime, cefotaxime, cotrimoxazole, gentamicin, minocycline, oxacillin and vancomycin). Fluoroquinolones inhibited essentially about 100% of H. influenzae, Salmonella and S. pneumoniae, more than 75% of Staphylococcus including methicillin-resistant strains, and about 90% of Enterobacteriaceae and 50% of P. aeruginosa. Minimal inhibitory concentration values ranged from < 0.015 to > 32 micrograms/ml for Klebsiella, S. aureus and epidermidis, E. coli and P. aeruginosa; from < 0.015 to 2 micrograms/ml for Salmonella; from 0.03 to 16 micrograms/ml for Serratia; from < 0.015 to 1 microgram/ml for Haemophilus; and from 0.5 to 2 micrograms/ml for S. pneumoniae. Levofloxacin and to a lesser extent ofloxacin and ciprofloxacin, generally exhibited a greater activity than the other agents against both Gram-positive and Gram-negative bacteria. Regarding the distribution of resistant strains in Italy, we found a peculiar pattern of resistance as far as E. coli and P. aeruginosa were concerned. Quality control parameters are also summarized. S. epidermidis resulted as a new emergent pathogen especially in immunocompromised patients and its level of sensitivity has been modified over the last few years. In fact, the percentage of resistant strains to antibiotics or the percentage of methicillin-resistant isolates (in our study 35%), has gradually increased. Levofloxacin and ofloxacin showed good activity against staphylococcal strains compared with the majority of other antibiotics. These results suggest that the newer quinolones are promising antimicrobial agents for various infections.     

Rapid detection of different RNA respiratory virus species by multiplex RT-PCR: application to clinical specimens

Inconsistent findings from recent mortality studies of workers exposed to magnetic fields have led to calls for more detailed understanding of exposure distributions and metrics in various industries. The authors undertook personal monitoring at an automobile transmission plant to (a) learn if magnetic field exposure differences were present, (b) make assignments for a brain cancer study, and (c) compare two exposure indices. A wide range of average exposures occurred (i.e., 0.016-4.6 microtesla). Within-day variability was also large, and it reached 4 orders of magnitude for some workers. Unexpectedly, demagnetizers were found among the strong sources that contributed to elevated exposures. The authors used conventional summary measures to assign job groups to exposure categories, and they used a new index of exposure irregularity to make alternative assignments. These new assignments appeared to differ from the original ones with respect to work time in each exposure group (i.e., 54% of the work time fell into different exposure categories).     

Rapid detection and typing of herpes simplex virus DNA in clinical specimens by the hybrid capture II signal amplification probe test

A second-generation signal amplification, nucleic acid-based test for the rapid detection and typing of herpes simplex virus (HSV) DNA was developed and evaluated with artificial and clinical specimens. The analytical sensitivity of the Hybrid Capture II (HC II) HSV DNA assay was determined by testing either cloned HSV DNA or total genomic HSV DNA titrations and resulted in detection thresholds of between 5 x 10(3) and 1 x 10(4) copies per assay. Specificity was assessed by testing a panel of bacteria and viruses commonly found in the female genital tract. Sensitivity was assessed by testing 112 ulcerative genital lesions by the HC II assay and comparing the results to those obtained by routine cell culture. Discrepant results were resolved by PCR testing. After resolution of the discrepant results, the sensitivity of the HC II assay compared to the consensus result (the results of two of three tests, the HC II assay, culture, and PCR, were in agreement) was 93.2% (41 of 44 specimens), and the specificity was 100% (60 of 60 specimens). Culture gave a sensitivity of 84.1% (37 of 44 specimens) and a specificity of 100% (60 of 60 specimens) compared to the consensus result. The results of HSV typing by the HC II assay and culture agreed in all cases. The HC II assay is a rapid and accurate assay for detecting and typing HSV types 1 and 2, with a sensitivity comparable to that of culture and greater ease of use than culture.     

Controlled clinical evaluation of Isolator and ESP aerobic blood culture systems for detection of bloodstream infections

A controlled clinical evaluation comparing the Isolator system (Wampole Laboratories, Cranbury, N.J.) and the ESP 80A blood culture bottle in the automated ESP system (Difco Laboratories, Detroit, Mich.) was performed with 10,535 blood culture sets from patients with suspected septicemia. Of 1,150 positive cultures, 844 positive cultures from 285 patients with 394 septic episodes fulfilled the study criteria for minimum blood sample requirements in each system and clinical significance of isolates. The Isolator system detected statistically significantly more positive cultures of Staphylococcus aureus (P < 0.001), Enterococcus spp. (P = 0.007), Escherichia coli (P = 0.001), Alcaligenes xylosoxidans (P = 0.02), Xanthomonas maltophilia (P = 0.01), Candida albicans (P < 0.001), and Candida glabrata (P = 0.05). The Isolator system detected significantly more septic episodes due to S. aureus (P < 0.001), X. maltophilia (P = 0.02), and C. albicans (P = 0.004) than did the ESP 80A bottle; however, the two systems did not otherwise significantly differ in their abilities to detect septic episodes due to other organisms.     

Rapid detection and serotyping of adenovirus by direct immunofluorescence

Four fluorescent antibody reagents were evaluated for their suitability for the identification of adenovirus isolates by immunofluorescence. The antibodies used in the reagents consist of monoclonal antibodies against adenovirus type 3 (Ad3), Ad4, Ad8, and adenoviruses of subgroup C (Ad1,2,5,6), serotypes known to occur in outbreaks of disease. Most of the monoclonal antibodies employed were reactive against type-specific antigens found on the hexon protein. Reagents employing two noncompeting anti-hexon antibodies were more sensitive than reagents prepared with only one monoclonal antibody, although both types of reagents exhibited a high degree of specificity. Five hundred and seventeen adenovirus isolates (359 of which had previously been typed by other methods) and 46 nonadenovirus isolates were examined with all four type-specific reagents in parallel with an adenovirus group-specific reagent. The results indicate that direct typing of adenovirus isolates is feasible, leading to significant savings in time compared to other typing methods and should contribute to the management of certain adenovirus infections, particularly during outbreaks.     

Nucleotide sequence analysis of a transposon (Tn5393) carrying streptomycin resistance genes in Erwinia amylovora and other gram-negative bacteria

A class II Tn3-type transposable element, designated Tn5393 and located on plasmid pEa34 from streptomycin-resistant strain CA11 of Erwinia amylovora, was identified by its ability to move from pEa34 to different sites in plasmids pGEM3Zf(+) and pUCD800. Nucleotide sequence analysis reveals that Tn5393 consists of 6,705 bp with 81-bp terminal inverted repeats and generates 5-bp duplications of the target DNA following insertion. Tn5393 contains open reading frames that encode a putative transposase (tnpA) and resolvase (tnpR) of 961 and 181 amino acids, respectively. The two open reading frames are separated by a putative recombination site (res) consisting of 194 bp. Two streptomycin resistance genes, strA and strB, were identified on the basis of their DNA sequence homology to streptomycin resistance genes in plasmid RSF1010. StrA is separated from tnpR by a 1.2-kb insertion element designated IS1133. The tnpA-res-tnpR region of Tn5393 was detected in Pseudomonas syringae pv. papulans Psp36 and in many other gram-negative bacteria harboring strA and strB. Except for some strains of Erwinia herbicola, these other gram-negative bacteria lacked insertion sequence IS1133. The prevalence of strA and strB could be accounted for by transposition of Tn5393 to conjugative plasmids that are then disseminated widely among gram-negative bacteria.     

Rapid sequence evolution of the mammalian sex-determining gene SRY

In mammals, induction of male sex determination requires the Y-chromosome gene SRY. SRY encodes a protein with a central 'high mobility group' domain (HMG box) of about 78 amino acids. HMG boxes are found in a wide variety of proteins that bind to DNA with high affinity but differing degrees of sequence specificity. The human SRY protein binds to linear DNA with sequence specificity and to cruciform DNA structures without sequence specificity. The DNA-binding activity of the SRY protein resides in the HMG box and mutations in this region are associated with sex reversal in XY females. No function has been ascribed to the portions of the SRY protein outside the HMG box. SRY belongs to a family of genes that are related by sequence homology within the DNA-binding domain: the genes most similar to SRY (> 60%) have been named SOX genes (SRY box genes). None of the known SOX genes is homologous to SRY outside the HMG-box region. Although SRY is an important developmental regulator, its sequence is poorly conserved between species apart from the HMG-box domain. Here we investigate the coding sequence of SRY in primates and find that evolution has been rapid in the regions flanking the conserved domain. The high degree of sequence divergence and the frequency of non-synonymous mutations suggest either that the majority of the coding sequence has no functional significance or that directional selection has occurred.     

Note on clinical judgment and the formal characteristics of clinical tasks.

Analysis of clinical judgment studies by F. J. Todd (1954), K. R. Hammond et al (see record 1965-08022-001), S. Oskamp (see record 1968-02701-001), L. R. Goldberg (see record 1970-12828-001), and R. M. Dawes (see record 1971-25701-001) suggests that the same relation between consistency of inferences and task predictability holds in clinical inferences as in laboratory learning studies. Findings indicate that, to understand the process of clinical inference, it is insufficient to analyze only the clinician; it is also necessary to analyze the clinician/clinical-task system. (15 ref) (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Rapid detection of species of the opportunistic yeast Trichosporon by PCR

Trichosporon species are opportunistic pathogens, associated with a high mortality rate in immunocompromised patients. Oligonucleotide primers were used to amplify a 170-bp fragment of small-subunit ribosomal DNA of all species in the genus Trichosporon by PCR. The primers amplify DNAs of all species in the genus Trichosporon, including six causative agents of trichosporonosis. DNAs of other medically important yeasts, such as Candida albicans and Cryptococcus neoformans, are not amplified by this detection system.