Evaluation of microscopic examination for bacteriuria.

In order to find an easily available, simple, reliable and inexpensive method for demonstrating significant bacteriuria in routine urine examination, microscopic observation and bacteriological cultures have been made in parallel on total of 206 urine samples. Microscopic examinations of centrifuged deposit for both pus cells and bacteria were found to be more satisfactory in urine specimens with significant bacteriuria than the examinations for either of these elements alone. The criteria of more than five pus cells per high power field and organisms visible in methylene blue stain had sensitivity of 79% and a false positive rate of 13%.     

Early detection and treatment of vesicular adenitis in bulls

The objective of this study was to determine whether early detection and treatment with tilmicosin would reduce the number of yearling bulls culled due to vesicular gland disease at the time of their first semen test. Bulls (n=2207) of 15 breeds at 17 performance test stations were examined by transrectal palpation at 9-12 months of age and assigned to three treatment groups: (1) positive treated group, receiving subcutaneous injections of tilmicosin every second day for three treatments (2) positive untreated control group, (3) negative untreated control group. Transrectal palpation of the glands was done again at a pre-sale semen test 28-70 d (mean=42.8 d) after the first examination. Semen was evaluated for the presence of pus and/or leukocytes by light microscopy. The proportion of bulls with vesiculitis was 4.4% (97/2207). At the pre-sale semen test the number of bulls with vesiculitis had decreased to 1.3% (29/2207); however, seven of these were new cases that developed after the first examination. Therefore, there was a recovery rate of 75/97 (77.3%) in the original group of bulls positive for vesiculitis. At the pre-sale test, there was no difference in the proportion of bulls with enlarged glands in the positive treated group (15/66) versus the positive untreated group (7/31) and no difference in the proportion of bulls with leukocytes or pus in their semen samples. The results of this experiment did not indicate any advantage in detection and treatment of vesiculitis before a year on age due to a high rate of spontaneous recovery.     

The utility of assessing the gross appearances of FNA specimens

F. Mayall, A. Cormack, S. Slater and K. McAnulty The utility of assessing the gross appearances of FNA specimens Objective: Ideally, fine needle aspiration (FNA) cytology should be performed with near‐patient assessment of the adequacy of the specimen by a cytopathologist. However, this is often not feasible. A cruder alternative is for the FNA practitioner to examine the gross appearances of the specimen and to try to predict the its quality. This study set out to determine the value of this approach. Methods: The study was conducted in tertiary public hospitals in New Zealand and the UK. FNA gross material grading was performed by a variety of pathologists on FNA samples taken using manual guidance and image guidance. The FNA gross material grade was compared with the findings on microscopic examination. Results: Nine out of 123 FNA samples were assessed as Grade 1 (unlikely to contain diagnostic material). All were subsequently reported as having insufficient diagnostic tissue on microscopic examination. Forty‐two of the FNA samples were assessed as Grade 2 (possibly contains diagnostic material) and 46 as Grade 3 (probably contains diagnostic material). None from either of these grades was reported as showing insufficient diagnostic material on microscopic examination. Twenty‐six cases were reported as Grade 4 (material suggesting a specific diagnosis). None of these was reported as showing insufficient diagnostic material on microscopic examination. The most common Grade 4 provisional diagnosis was that of a colloid cyst or colloid nodule of the thyroid (seven cases). Only two cases had misleading Grade 4 provisional diagnoses. Both were thought to be pus on gross examination but showed necrotic carcinoma on microscopic examination. Conclusions: The gross appearances of FNA samples can usually predict the adequacy of the samples and sometimes predict the final microscopic diagnosis. However, near‐patient microscopic assessment of FNA specimens is preferable if available.     

Preparation of Drosophila mitotic chromosomes for electron-microscopic studies]

A method of chromosome spreading on microscopic slides was modified for electron microscopy of metaphase chromosomes in Drosophila tissues. The slides covered with an electron transparent film were plasmochemically modified to make them hydrophilic. A piece of fixed tissue was macerated in 60% propionic acid before spreading chromosomes over the slide. The parts of preparation selected under light microscope for electron microscopic examination were cut and peeled of the slide to the top of a water drop. It was shown that the resolution of chromosomal structures was significantly higher than seen under optical microscope, but lower than in serial sections.     

Effect of the storage period of paraffin sections on the detection of mRNAs by in situ hybridization.

In this study we evaluated whether storing non-deparaffinized sections can affect the detection of specific mRNAs by radioactive in situ hybridization (ISH). Using a standard ISH protocol, we hybridized serial sections of paraffin blocks stored for different periods of time with (33)P-labeled riboprobes specific for rat Type III collagen and matrix metalloproteinase-2 (MMP-2). Signal intensities were evaluated using a phosphorimager and by blinded microscopic examination. For slides hybridized with the Type III collagen riboprobe, signal intensities measured with the phosphorimager or evaluated by microscopic examination were negatively correlated with the storage period of the sections. For slides hybridized with the MMP-2 riboprobe, differences in signal intensity could be detected, albeit inconsistently, with the phosphorimager, although microscopic examination consistently indicated stronger signals in freshly sectioned slides compared to slides stored for 2 weeks or more. We concluded that it was preferable to use recently prepared sections for trying to locate mRNAs in paraffin-embedded tissues by ISH. In addition, our results suggest that quantifying signal intensity using a phosphorimager is feasible for abundant mRNAs or when large differences in expression are anticipated.(J Histochem Cytochem 49:927-928, 2001)     

Bacteriological quality assessment of fresh marketed lettuce and fennel.

One-hundred and twenty samples of lettuce and 89 samples of fennel purchased from five retail outlets in the city of Bari (Italy) from October 1973 through September 1975 were examined for viable aerobic bacteria (AB), total coliforms (TC), fecal coliforms (FC), fecal streptococci (FS), and salmonellae. Comparative tests indicated that the results of bacteriological analysis of wash water from either vegetable by a membrane filter technique compared favorably with those of conventional cultural examination of the vegetable tissue for the purpose of providing an indication of the bacteriological quality of the samples. Using the membrane filter technique, 2-year average counts of 6.59 X 10(7) for AB, 5.95 X 10(4) for TC, 6.13 X 10(3) for FC and 2.24 X 10(3) for FS/100 g (fresh weight) were obtained with lettuce; with fennel, the corresponding figures were 2.32 X 10(6) for AB, 7.82 X 10(4) for TC, 7.8 X 10(4) for FC, and 3.15 X 10(3) for FS. Indicator bacteria were present in all samples examined. In addition, 68.3% of the lettuce and 71.9% of the fennel samples yielded one or more of the following Salmonella serotypes: S. schottmuelleri, S. typhimurium, S. thompson, S. dublin, S. typhi, and S. anatum.     

Effect of ascorbic acid prophylaxis on the frequency of chromosome aberrations, urine mutagenicity and nucleolus test in workers occupationally exposed to cytostatic drugs

The genetic risk of workers occupationally exposed to a series of newly developed cytostatic drugs and the presumed antimutagenic potential of ascorbic acid (AA) were studied in a group of 38 chemical laboratory personnel examined for chromosome aberrations in lymphocytes, urine mutagenicity and nucleolar RNA activity before and after a 6-month prophylactic administration of AA at daily doses of 1 g for 5 days a week. Chromosome aberration tests revealed elevated aberrant cell (AB.C) rates both prior to and after AA supplementation (3.9% and 3.65% of AB.C., respectively). These values were significantly higher than those found in 18 non-exposed matching controls (1.05% of AB.C.). Tests for mutagenic activity in the urine of drug-exposed workers revealed 64% positive urine samples prior to vitaminization and 60% positive urine specimens after it; positive urine samples in the group of controls accounted for 21% of samples. In the nucleolus test, numbers of inactivated micronuclei in the exposed were initially higher than those of controls (33.4% versus 24.3%), but dropped to 20.5% after AA supplementation. These findings show that AA prophylaxis alone cannot substantially reduce the hazards associated with exposure to anti-cancer drugs.     

Ziehl-Neelsen staining technique can diagnose paragonimiasis

Background

We evaluated the Ziehl-Neelsen staining (ZNS) technique for the diagnosis of paragonimiasis in Laos and compared different modifications of the ZNS techniques.

Methodology

We applied the following approach: We (1) examined a paragonimiasis index case''s sputum with wet film direct examination (WF) and ZNS; (2) re-examined stored ZNS slides from two provinces; (3) compared prospectively WF, ZNS, and formalin-ether concentration technique (FECT) for sputum examination of patients with chronic cough; and (4) compared different ZNS procedures. Finally, we assessed excess direct costs associated with the use of different diagnostic techniques.

Principal Findings

Paragonimus eggs were clearly visible in WF and ZNS sputum samples of the index case. They appeared brownish-reddish in ZNS and were detected in 6 of 263 archived ZNS slides corresponding to 5 patients. One hundred sputum samples from 43 patients were examined with three techniques, which revealed that 6 patients had paragonimiasis (13 positive samples). Sensitivity per slide of the FECT, ZNS and the WF technique was 84.6 (p = 0.48), 76.9 (p = 0.25) and 61.5% (p = 0.07), respectively. Percentage of fragmented eggs was below 19% and did not differ between techniques (p = 0.13). Additional operational costs per slide were 0 (ZNS), 0.10 US$ (WF), and 0.79 US$ (FECT). ZNS heated for five minutes contained less eggs than briefly heated slides (29 eggs per slide [eps] vs. 42 eps, p = 0.01). Bloodstained sputum portions contained more eggs than unstained parts (3.3 eps vs. 0.7 eps, p = 0.016).

Conclusions/Significance

Paragonimus eggs can easily be detected in today''s widely used ZNS of sputum slides. The ZNS technique appears superior to the standard WF sputum examination for paragonimiasis and eliminates the risk of tuberculosis transmission. Our findings suggest that ZNS sputum slides should also be examined routinely for Paragonimus eggs. ZNS technique has potential in epidemiological research on paragonimiasis.     

Frequency of Strongyloides stercoralis infection in alcoholics

Several studies have shown that chronic alcoholics have increased susceptibility to infections due to higher exposure to infectious agents as well as breakdown in their immune defenses. As Strongyloides stercoralis infection is usually more relevant in immunocompromised patients, the aim of this study was to evaluate the frequency of S. stercoralis infection in alcoholics. Thus, coproparasitological examination was carried out in 145 subjects, from which 45 were chronic alcoholics (mean age of 45.7 +/- 11.0 years), 10 were nonalcoholic cirrhotic patients (mean age of 50.2 +/- 13.1 years), and 90 were asymptomatic nonalcoholic subjects (mean age of 46.7 +/- 10.1 years), which served as controls. From the alcoholics, 9 had hepatic cirrhosis, 9 had chronic pancreatitis and 27 had neither cirrhosis nor pancreatitis. For the diagnosis of strongyloidiasis, the Baermann-Moraes and Lutz methods were used in three fecal samples from each subject. Samples were collected at alternated days, and three slides of each sample were analyzed for each method, thus totalizing 2,610 slides examined. The frequency of strongloidiasis in the total alcoholic group (33.3%) and in the subgroups of alcoholics, i.e., patients with hepatic cirrhosis (44.4%), with chronic pancreatitis (33.3%), and those with no cirrhosis or pancreatitis (29.6%) was statistically higher than that found in the control group (5.5%). None of the individuals with nonalcoholic hepatic cirrhosis had S. stercoralis infection. Our results showed that the chronic alcoholism itself is an important factor that predisposes to strongyloidiasis.     

Limitations of detection of malignancy in pleural effusions using ELISA-based TRAP assay: comparison with cytological examination

OBJECTIVE: Telomerase is active in almost all cancers from various organs but is not detectable in most normal cells. Thus, telomerase activity might be a universal and specific marker for diagnosing malignancy. The aim was to evaluate the potential use of the ELISA-based TRAP assay to detect malignancy in pleural effusion, and to compare it with conventional cytological examination. METHODS: Using the ELISA-based TRAP assay, telomerase activity was examined in 94 consecutive pleural effusions submitted for cytological examination. RESULTS: According to the results of cytology, the 94 samples were divided into two groups: group I, 79 non-malignant pleural effusions, including group IA, no association with a malignant tumour, a control group (n = 63), and group IB, associated with a malignant tumour (n = 16); and group II, 15 malignant pleural effusions. Telomerase activity was detected in five of 63 samples in group IA (7.9%), four of 16 samples in group IB (25%), and six of 15 samples in group II (40%). All five false-positive effusions were from patients with tuberculosis. Comparing group II with group IA, the TRAP assay showed 40% sensitivity, 92.1% specificity, 54.5% positive and 86.6% negative predictive value, and 82.1% accuracy. However, the detection rate of the TRAP assay (88.9%) was higher than that of the cytological examination (66.7%) in lung cancer-inflicted pleural effusions. CONCLUSION: The ELISA-based TRAP assay is relatively insensitive; therefore, it is unsuitable as a routine diagnostic tool for pleural effusion. False-positive telomerase activity due to lymphocytic contamination may weaken its diagnostic value for malignant effusions in a tuberculosis-endemic area.     

EVALUATION OF A NEW METHOD OF DUST CONTROL

Microscopic dust sampling was done to determine the amount of dust in the homes of six patients who were sensitive to house dust and who had allergic disease that was intractable to treatment. One case was subsequently excluded from the study because of extraordinary circumstances. The remaining five cases were studied with repeated dust counts before and after a water-and-oil emulsion was sprayed in the patient''s bedroom to immobilize house dust. In all five cases, the patients had dramatic relief of symptoms after the spraying was done. In four out of five, there was concomitant reduction of the amount of dust in the air as determined by microscopic counting of the dust particles on a glycerincoated slide. In the fifth case, relief of symptoms was not accompanied by reduction of dust on slides, but investigation revealed an error in control of exposure of the slides.     

Direct Immunofluorescence Test for the Diagnosis of Genital Herpesvirus Infections

Herpetic lesions of the genitalia may be confused clinically with other ulcerative, genital lesions. Direct immunofluorescence (FA) provides a rapid method of diagnosis, and the utility of this method for the diagnosis of genital ulcers was examined. One hundred and ten patients with genital lesions were examined by darkfield for syphilis and by FA and culture for herpes simplex virus (HSV) infections. Satisfactory samples were obtained from 102 patients, of which 81 were clinically suspected cases of HSV. Acetone-fixed slides of scrapings of ulcerative lesions were stained with conjugated antiserum prepared in rabbits against HSV type 2. HSV was isolated from 73% of specimens of suspected herpetic lesions, and 77% of these specimens were positive by FA. Nine percent were positive by FA only and these were not thought to represent false positives. Five percent were positive by culture only. A comparison of clinical diagnoses with laboratory findings revealed that 4% of the cases were misdiagnosed when only the clinical evaluation was considered. The data suggest that the inclusion of a diagnostic FA test for HSV along with the darkfield examination may be useful for differentiating the etiological agents of ulcerative, genital lesions.     

Gram staining with an automatic machine

This study was undertaken to develop a new Gram-staining machine controlled by a micro-controller and to investigate the quality of slides that were stained in the machine. The machine was designed and produced by the authors. It uses standard 220 V AC. Staining, washing, and drying periods are controlled by a timer built in the micro-controller. A software was made that contains a certain algorithm and time intervals for the staining mode. One-hundred and forty smears were prepared fromEscherichia coli, Staphylococcus aureus, Neisseria sp., blood culture, trypticase soy broth, direct pus and sputum smears for comparison studies. Half of the slides in each group were stained with the machine, the other half by hand and then examined by four different microbiologists. Machine-stained slides had a higher clarity and less debris than the hand-stained slides (p<0.05). In hand-stained slides, some Gram-positive organisms showed poor Gram-positive staining features (p<0.05). In conclusion, we suggest that Gram staining with the automatic machine increases the staining quality and helps to decrease the work load in a busy diagnostic laboratory.     

Streptococci species of anginosus group isolated from oral and maxillofacial infections

The aim of the study was to isolate and identify at species level streptococci strains of anginosus group in pus samples collected from 110 patients with oral and maxillofacial (OMF) infections. Gram-stained smears and cultures on selective and nonselective media were done from each of the 111 pus samples (2 samples were collected from one of the patients, who presented 2 oral abscesses at the same time). The isolates were identified on the basis of cultural and biochemical characteristics. Speciation of the anginosus group isolates was performed using the Rapid ID 32 Strep system (Bio Mérieux, France). Fourty-four anginosus group strains were isolated from 42 patients. Fourty of these isolates were identified as Streptococcus anginosus (2 nonidentical isolates were found in 2 patients), 3 isolates as Streptococcus constellatus and only one as Streptococcus intermedius. The study confirmed that the anginosus group is often involved in OMF infections alone or in association with other aerobic and/or anaerobic bacteria. In the investigated cases, Streptococcus anginosus was by far the most frequently isolated species within the anginosus group.     

Diagnostic performance of mycologic and serologic methods in a cohort of patients with suspected sporotrichosis

BackgroundThe gold standard for the sporotrichosis diagnosis is culture; however, serologic approaches have been recently implemented to aid in the sporotrichosis diagnosis. Nevertheless, the clinical consequences of the introduction of serologic tests are poorly addressed.AimsTo correlate the results of culture and serology of patients with suspected sporotrichosis.MethodsA retrospective study of 198 patients with suspected sporotrichosis was conducted. Information about culture isolation of Sporothrix from clinical samples and antibody detection by an enzyme-linked immunosorbent assay (ELISA) were obtained from the medical records of the patients.ResultsPositive culture and antibody detection was observed in the samples of 84 patients (42.4%). Forty-one samples (20.7%) showed negative results with both techniques and divergent results were obtained in the samples of 73 patients (36.9%). False negative results in the ELISA were observed with 23 patients (31.5%), 78.3% of them with less than 30 days of infection (p = 0.0045). Among the initial false positive ELISA in the sera of 50 patients, four samples in culture yielded the growth of Sporothrix, and 27 improved with itraconazole. At the end of follow-up, a diagnosis of proven or probable sporotrichosis was established in 139 patients, and possible sporotrichosis in 11 patients. The treatment of the patients with probable sporotrichosis with antifungal drugs resulted in clinical cure for these individuals.ConclusionsThese two techniques are complementary in the diagnosis of sporotrichosis, making diagnosis and clinical decision more precise.     

Entamoeba histolytica and/or Entamoeba dispar: infection frequency in HIV+/AIDS patients in Mexico city

The objective of this work was to evaluate the frequency of Entamoeba histolytica/Entamoeba dispar intestinal infection in HIV+/AIDS subjects and their HIV- close relatives or sexual partners. Enteric parasites were investigated in stool samples by microscopic examination and E. histolytica and E. dispar were identified by PCR. We found by microscopic analysis in HIV+/AIDS group that the E. histolytica/E. dispar complex was present in 5.9% of the members, while in the HIV- group was 2.9%. With PCR we found that the E. histolytica prevalence was 25.3% in the HIV+/AIDS group and 18.5% in the HIV-group. The difference in the results obtained with the microscopic and PCR is due to the different sensibility of the procedures. Besides, we found patients who were infected with E. histolytica in both groups were asymptomatic cyst passers. Our results suggest that E. histolytica strains prevalent in the studied community appear to be of low pathogenic potential.     

Human papillomavirus infection of the uterine cervix of women without cytological signs of neoplasia.

One hundred and six patients were studied whose cervical smears showed only non-specific inflammatory changes. Screening for genital pathogens yielded only a few positive cases. Histological examination of biopsy specimens taken by colposcopically directed tissue sampling showed cervical intraepithelial neoplasia in 13 of the women (12.3%). Deoxyribonucleic acid (DNA) hybridisation techniques were used to detect human papillomavirus, which was found in 24 patients (22.6%). In a second group of 104 patients with normal cervical cytology tissue biopsy samples were obtained and examined histologically but in no case was cervical intraepithelial neoplasia found. On DNA hybridisation, however, 12 patients (11.5%) were found to be positive for human papillomavirus. In this group finding human papillomavirus DNA was usually associated with a columnar ectopy. An association between human papillomavirus type 16 DNA and both cervical intraepithelial neoplasia and cervical cancer is well established. In this study it was type 16 which occurred most frequently in both groups.     

Negative micronucleus tests on caprolactam and benzoin in ICR/JCL male mice

Male ICR/JCL mice were given a single intraperitoneal injection of 125, 250, and 500 mg/kg of caprolactam (CAP), or 250, 500, and 1000 mg/kg of benzoin (ZOIN). Bone marrow preparations were made 24, 30, and 48 h after treatment with the maximum dose, and 30 h after treatment with the other doses. The slides were coded before microscopic examination. No significant increase was found in the incidence of micronucleated polychromatic erythrocytes after treatment with either CAP or ZOIN.     

Evaluation of two coproscopic techniques for the diagnosis of schistosomiasis in a low-transmission area in the state of Minas Gerais, Brazil

This population study, which evaluated two parasitological methods for the diagnosis of schistosomiasis mansoni, was performed in a low-transmission area in Pedra Preta, Montes Claros, Minas Gerais, Brazil. A total of 201 inhabitants of the rural area participated in this research. Four stool samples were obtained from all participants and analysed using the Kato-Katz method (18 slides) and a commercial test, the TF-Test?, which was performed quantitatively. The data were analysed to determine prevalence, the sensitivity of the diagnostic methods, the worm burden and the definition of the "gold standard", which was obtained by totalling the results of all samples examined using the Kato-Katz technique and the TF-Test?. The results showed that the prevalence obtained from the examination of one Kato-Katz slide (the methodology adopted by the Brazilian control programme) was 8% compared to 35.8% from the "gold standard", which was a 4.5-fold difference. This result indicates that the prevalence of schistosomiasis in so-called low-transmission areas is significantly underestimated.     

谷红注射液和丁苯酞注射液联合Solitaire AB型支架取栓治疗急性缺血性脑卒中的临床研究

摘要 目的：探讨谷红注射液和丁苯酞注射液联合Solitaire AB型支架取栓治疗急性缺血性脑卒中（AIS）的临床疗效。方法：选择2018年1月到2020年12月期间攀枝花市中西医结合医院收治的AIS患者98例,依照随机数字表法分为观察组（50例）和对照组（48例）。对照组采用Solitaire AB型支架取栓治疗,观察组给予谷红注射液和丁苯酞注射液联合Solitaire AB型支架取栓治疗,两组均治疗14 d。治疗后进行临床疗效评价,比较两组治疗后的血管再通率和并发症发生率,比较两组治疗前后的美国国立卫生研究院卒中量表（NIHSS）评分、Barthel指数评分及全血高切黏度、全血低切黏度、血小板聚集率。结果：观察组的临床总有效率为84.00%（42/50）,对照组为60.42%（29/48）,两组比较差异有统计学意义（P<0.05）。观察组的血管再通率明显高于对照组（P<0.05）。两组治疗后血小板聚集率、全血低切黏度及全血高切黏度均明显低于治疗前（P<0.05）,且观察组治疗后血小板聚集率、全血低切黏度及全血高切黏度均明显低于对照组（P<0.05）。治疗后观察组Barthel指数评分明显高于对照组（P<0.05）,NIHSS评分明显低于对照组（P<0.05）。观察组的并发症发生率为6.00%（3/50）,对照组为4.17%（2/48）,两组比较差异无统计学意义（P>0.05）。结论：谷红注射液和丁苯酞注射液联合Solitaire AB型支架取栓治疗AIS的临床疗效显著,能够提高血管再通率,减轻神经功能缺损,改善患者的日常生活能力和血液流变学指标,且并发症较轻。