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PubMed1997

Bayes estimates for immunological progression rates in HIV disease.

We develop Bayesian methods for calculating shrinkage estimates of immunological progression rates (for example, CD4 count decline rates) in populations of HIV-infected patients. These methods make the assumption that decline of immunological markers may be modelled as approximately linear on some suitable chosen scale. They are applicable in situations where seroconversion times are unknown and follow-up of patients is variable, with some patients having only sparse measurements of immunological markers. Fitting of models is achieved by Gibbs sampling and CD4 count data from 603 members of the Edinburgh City Hospital Cohort with at least two CD4 determinations are analysed to provide an illustration. It is found that Bayesian shrinkage estimates for CD4 slopes on the square root scale are much more effective predictors of future CD4 counts than the least squares estimates, with respect to squared error loss. Of various shrinkage estimators considered, the most effective corresponds to the simplest model, which can also be fitted using SAS. A characterization of the pattern of CD4 loss in the Edinburgh cohort is obtained (mean rate of decline on root scale-1.61 per annum, standard deviation 1.03) and the effect of various covariates (sex, age, risk category and HLA antigen type) on immunological progression is considered. It is found that homosexual men in Edinburgh and patients with HLA haplotype A1B8DR3 experience significantly faster loss of CD4.

Adult↗
PubMed1983

Suppression of the periodontopathic microflora in localized juvenile periodontitis by systemic tetracycline.

Since recent studies have implicated Actinobacillus actinomycetemcomitans in the etiology of localized juvenile periodontitis, this investigation determined the effectiveness of subgingival debridement, topical Betadine Solution, and systemic tetracycline in suppressing subgingival A. actinomycetemcomitans and other microorganisms. A total of 20 deep periodontal pockets and 10 normal periodontal sites of 6 localized juvenile periodontitis patients was included in the study. Each patient was treated in 3 stages over a period of 22 weeks, and the result of treatment was monitored for an additional 38 weeks. The first stage of treatment included plaque control, as well as thorough scaling and root planing, composed of at least 6 h of debridement. No concomitant periodontal surgery was performed. In the second stage, Betadine saturated cotton gauze was inserted into the periodontal pockets for 10 min. Stage 3 involved systemic tetracycline therapy (1 g/day) for 14 days. The subgingival microflora was determined at frequent intervals by selective culturing of A. actinomycetemcomitans and Capnocytophaga and by direct microscopic examination. The clinical effect was assessed by measuring changes in probing periodontal attachment level, probing periodontal pocket depth, radiographic alveolar bone mass, and other relevant clinical parameters. Scaling and root planing reduced the total subgingival bacterial counts and the proportions of certain Gram-negative bacteria, but no periodontal pocket became free of A. actinomycetemcomitans. Betadine application had little or no effect on the subgingival microflora. In contrast, tetracycline administered via the systemic route suppressed A. actinomycetemcomitans, Capnocytophaga, and spirochetes to low or undetectable levels in all test periodontal pockets. A. actinomycetemcomitans reappeared in 9 of the deep periodontal pockets after the administration of tetracycline. Most of these 9 pockets became free of detectable A. actinomycetemcomitans during the second week of tetracycline administration, whereas pockets which yielded no A. actinomycetemcomitans after tetracycline therapy became free of the organisms during the first week of tetracycline treatment. This data suggests that systemic tetracycline therapy of localized juvenile periodontitis should, as a practical rule, be continued for 3 weeks. Periodontal destruction continued in 4 deep pockets which all showed high posttetracycline A. actinomycetemcomitans counts. All 6 pockets which demonstrated a marked gain in periodontal attachment yielded no cultivable A. actinomycetemcomitans. No association was found between periodontal disease status and subgingival Capnocytophaga, spirochetes or motile rods. The present study indicates that A. actinomycetemcomitans is an important etiologic agent in localized juvenile periodontitis.(ABSTRACT TRUNCATED AT 400 WORDS)

Actinobacillus↗
PubMed2001

Impaired whole-blood polymorphonuclear leukocyte migration as a possible predictive marker for infections in preterm premature rupture of membranes.

OBJECTIVES: Steroids, used in preterm premature rupture of membranes (pPROM), to reduce the risk of morbidity and mortality of the preterm neonate, impair the maternal polymorphonuclear leukocyte (PMN)-based immune system. In spite of combination with antibiotics, prenatal and postnatal bacterial infections of mother and child are frequent. This pilot study focuses on the influence of steroids in pPROM on maternal PMN functional capacity and subsequent infections. METHODS: After opting for expectant management, eight women with pPROM and no signs of infection were treated by steroids (betamethasone 5.7 mg, i.m. every 24 hours, for three days) and antibiotic therapy with either amoxicillin and clavulanic acid, piperacillin or ampicillin i.v. up to delivery. The conventional inflammation parameters of PMN blood count and C-reactive protein (CRP) were measured daily in parallel with PMN migratory capacity towards N-formyl-methionyl-leucyl-phenylalanine stimulation and under blank conditions, estimated by a whole blood membrane filter assay. RESULTS: In all patients PMN migration decreased during the application of steroids. Three patients showed a decrease in PMN migration below critical values and in spite of antibiotic prophylaxis acute pyelonephritis developed 2-6 days later. PMN count and CRP were not predictive of maternal infection. CONCLUSION: Reduced PMN function, caused by steroid treatment in pPROM, is suggested to be a reason for serious bacterial infections in spite of antibiotic prophylaxis. PMN migration reflects individual PMN defensive capacity.

Antibiotic Prophylaxis↗
PubMed1975

Proteolysis detection in milk. IV. Starch-gel electrophoresis and formol titration.

Starch-gel electrophoresis (SGE) and formol titration methods for detecting proteolysis in cold-stored raw milk have been studied to establish their value as quality indices. When examined by SGE, the first evidence of proteolysis in raw milks stored at 5 degrees C was the formation of para-k-casein. However, this fraction could not be detected on the starch gels until the total bacterial count (TBC) exceeded 10-minus 7/ml. The SGE method appeared more reliable than the previously discussed tyrosine value method. Formol titration did not appear to have any application in the screening of cold-stored raw milks with TBC less than 10-minus 7/ml, but would detect some milks of poorer bacteriological quality.

Animals↗
PubMed1992

Enhanced collagen phagocytosis by rat molar periodontal fibroblasts after topical application of lipopolysaccharide--ultrastructural observations and morphometric analysis.

To investigate the effect of lipopolysaccharide (LPS) on phagocytic activity of collagen fibrils by periodontal fibroblasts, we studied rat molar gingival connective tissue and periodontal ligament under light and electron microscopy after topical application of LPS (5 mg/ml in physiological salt solution (PS)) on the gingival sulcus. Phagocytic activity of collagen fibrils by fibroblasts was evaluated by counting the number of collagen-containing vacuoles inside fibroblasts that were present within a defined area (1200 microns2). Values obtained from fibroblasts in the subepithelial connective tissue, the region near the alveolar crest, and the middle region of periodontal tissue were compared. Periodontal ligament fibroblasts showed increased phagocytosis of the collagen fibrils from 3 hours to 1 day after topical LPS application, but no differences were observed in the gingival tissue. The intracytoplasmic vacuoles containing collagen fibrils were of various sizes and shapes, showing positive for acid phosphatase and/or alkaline phosphatase reaction. Collagen phagocytic activity of the fibroblasts in the middle region of the periodontal ligament also increased after PS treatment. However, this was significantly less than that observed in LPS-treated animals (p less than 0.01). This study indicates that LPS may enhance the degradation of collagen by stimulating the phagocytic activity of the periodontal ligament fibroblasts.

Animals↗
PubMed1990

[Variable threshold levels for estimation of renal uptake of 99mTc-dimercaptosuccinic acid based on single photon emission computed tomography].

To calculate renal uptake of 99mTc-dimercaptosuccinic acid (DMSA) more accurately using single photon emission computed tomography (SPECT), it is necessary to estimate values of threshold level, which corresponds to the ratio of kidney to background (BG ratio). Thus the phantom and clinical studies were conducted. Six kinds of renal phantoms of 80 to 339 ml, contained different radioactivity of 37 to 485 MBq were prepared. These phantoms were placed in a larger body phantom filled with 1 to 50% of radioactivity representing background. Clinical application of this method was also performed. Results were as follows. 1) A significant correlation between phantom volumes and estimated volumes obtained using threshold level based on BG ratio calculated on tomographic images was found (r = 0.99). 2) A significant correlation between total counts in estimated volumes and radioactivities in phantoms was recognized (r = 0.94). 3) Known radioactivity in phantoms and radioactivity in estimated volumes were highly related (r = 0.98). 4) Clinical application showed valuable results in patients with renal dysfunction. Thus, this method can calculate more accurate renal uptake of 99mTc-DMSA.

Adult↗
PubMed1999

Study on local inflammatory reactions and other parameters during subcutaneous mistletoe application in HIV-positive patients and HIV-negative subjects over a period of 18 weeks.

Subcutaneous injections of fermented and unfermented aqueous extracts of Viscum album L. result in a local inflammatory reaction at the injection site. In this trial, the symptoms associated with this local reaction were investigated. Furthermore the occurrence of local reactions was tried to correlate with an increase in CD3/25- and CD8/38-positive lymphocyte counts, with eosinophilic granulocyte numbers, and with the formation of mistletoe lectin antibodies. Included in the trial were 30 HIV-antibody-positive patients and 17 healthy non-smokers, aged 24-51 years. The CD4 cell count in the HIV-negative subjects was > 800/microliter, compared with 200-600/microliter in the HIV-positive patients. All study participants had a Karnofsky score > or = 70. The trial subjects were observed over a period of 18 weeks. With escalation of the dose of a fermented and unfermented extract of Viscum album L. (Iscador Qu Spezial and Viscum album QuFrF), there was an increase in local reactions. Erythema at the injection site was the most frequently reported symptom. Between the doses and the symptoms induration, swelling and pruritus were marked correlations. Effects of the application of mistletoe extracts on the immune system were demonstrated by an increase in CD3/25-positive lymphocyte counts and antibodies against mistletoe-lectins. There were no changes in eosinophilic granulocytes or CD8/38-positive lymphocyte populations. For evaluation of the therapeutic applications of mistletoe extracts in HIV-positive patients it is advisable to assess primarily activation of CD3-positive lymphocytes and the patient response on the basis of the local reaction. The local inflammatory reaction at the injection site is desirable and well tolerated if the reaction is smaller than 5 cm in diameter.

Adjuvants, Immunologic↗
PubMed2001

Eurosol versus fetal bovine serum-containing corneal storage medium.

PURPOSE: To evaluate the usability of Eurosol, a new medium-term corneal storage medium without components of bovine origin. METHODS: Ten pairs of human donor corneas were placed in tissue culture at 31 degrees C for 7, 14, 21, 28, or 35 days. One cornea of each pair was cultivated in conventional storage medium on Earls' minimum essential medium base containing 2% fetal bovine serum; the other one was stored in Eurosol. Corneas were examined with inverse light microscopy; corneal thickness was measured; and scanning electron microscopy was performed. RESULTS: No significant difference in corneal thickness and endothelial cell count was found at any time. Scanning electron microscopy showed a complete endothelial cell layer on all corneas. CONCLUSION. The findings indicate a potential clinical applicability of the tested serum-free medium-term storage medium, offering a safer alternative to conventional media containing fetal bovine serum.

Cell Count↗
PubMed1999

Quantification of eosinophilic granule protein deposition in biopsies of inflammatory skin diseases by automated image analysis of highly sensitive immunostaining.

Eosinophilic granulocytes are major effector cells in inflammation. Extracellular deposition of toxic eosinophilic granule proteins (EGPs), but not the presence of intact eosinophils, is crucial for their functional effect in situ. As even recent morphometric approaches to quantify the involvement of eosinophils in inflammation have been only based on cell counting, we developed a new method for the cell-independent quantification of EGPs by image analysis of immunostaining. Highly sensitive, automated immunohistochemistry was done on paraffin sections of inflammatory skin diseases with 4 different primary antibodies against EGPs. Image analysis of immunostaining was performed by colour translation, linear combination and automated thresholding. Using strictly standardized protocols, the assay was proven to be specific and accurate concerning segmentation in 8916 fields of 520 sections, well reproducible in repeated measurements and reliable over 16 weeks observation time. The method may be valuable for the cell-independent segmentation of immunostaining in other applications as well.

Automation↗
PubMed1989

The effect of citrus limonoids on hamster buccal pouch carcinogenesis.

Two citrus limonoids, limonin and nomilin, were tested for their effects on the development of 7,12-dimethylbenz[a]anthracene (DMBA)-induced buccal pouch epidermoid carcinomas. Forty-five female Syrian hamsters were divided into three equal groups. The left buccal pouches of the animals in each group were pretreated topically with two applications of dimethylsulfoxide (DMSO) (group I), a 2.5% solution of limonin dissolved in DMSO (group II) or a 2.5% solution of nomilin dissolved in DMSO (group III). After this initial treatment, 11 hamsters from each group were selected. The left buccal pouches of these animals were painted 2 or 3 times weekly with a 0.5% solution of DMBA in mineral oil. On alternate days the pouches were painted with DMSO (I), the 2.5% solution of limonin (II) or the 2.5% solution of nomilin (III). The 12 remaining hamsters were used as controls and were painted with mineral oil and DMSO (I), mineral oil and the 2.5% solution of limonin (II), or mineral oil and the 2.5% solution of nomilin (III). After 15 weeks the hamsters were killed, the pouches were excised and the tumors were counted and measured. Tumors of variable size were common in the animals treated with DMBA. However, the animals receiving topical applications of limonin exhibited a 60% reduction in tumor burden. Further comparisons between groups I and II showed that this reduction in tumor burden was due to a 20% decrease in tumor number and a 50% decrease in tumor mass. The results for group III showed that nomilin was considerably less effective as an inhibitor of DMBA-induced neoplasia.

9,10-Dimethyl-1,2-benzanthracene↗
PubMedDate not supplied

Comparison of two methods for presurgical disinfection of the equine hoof.

OBJECTIVES: To determine for equine hooves the normal resident aerobic bacterial population and the efficacy of 2 methods of disinfection. Study Design-Measurement of total bacterial, gram-positive bacterial, and gram-negative bacterial surface populations from the frog, sole, and hoof wall after each step of 2 different preoperative surgical disinfection techniques. ANIMALS: Six adult horses. METHODS: Hoof wall, sole, and frog samples were collected for quantitative bacteriology before, during, and after 2 multistep antiseptic preparation techniques: Method A-6-minute scrub with povidone-iodine soap, followed by 24-hour submersion in povidone-iodine solution-soaked cotton; and Method B-initial removal of superficial layer of hoof capsule before completing Method A disinfection procedures. RESULTS: Removal of the superficial hoof layer, application of the povidone iodine scrub, and completion of the povidone-iodine soak all significantly (P < .0008) decreased total bacterial numbers. Method B had significantly lower bacterial counts than method A at each consecutive step. Final total bacterial counts remained greater than 10(5) bacteria per gram of tissue regardless of preparation method. CONCLUSIONS: The hoof surface hosts a broad spectrum of aerobic gram-positive and -negative bacteria, many of which are potential pathogens. Bacterial numbers can be significantly reduced by removal of the superficial hoof surface, by application of a povidone-iodine scrub, and by use of a 24-hour povidone-iodine soak. However, bacterial populations >10(5) g per tissue persist after these disinfection procedures. CLINICAL RELEVANCE: Regardless of the preparation methods used in this study, bacterial populations capable of inducing wound infection remain on the hoof capsule.

Animals↗
PubMed1997

[Mitoxantrone-induced acute left heart failure after intrapleural administration].

A 79-year-old female was admitted to our hospital because of a malignant pleural effusion following mastectomy 4 years ago. In the patient's history arterial hypertension and previous inferior myocardial infarction have been known. Two doses of 20 mg mitoxantrone were installed intrapleurally at an interval of 4 weeks. Six hours after the second mitoxantrone application and the patient had increasing dyspnea with consecutive left heart failure, pulmonary congestion, and a drop of blood pressure. The white-cell count was 14800/mm3. The levels of creatinine phosphokinase (CPK), lactate dehydrogenase (LDH) and serum aspartate aminotransferase (SGOT) were in the normal range. Transthoracic echocardiography showed concentric left ventricular hypertrophy and a markedly decreased fractional shortening, but no left ventricular dilatation. The electrocardiogram showed newly appeared down-sloping ST-segments and inverted T-waves. Clinical recovery was achieved after 6 days by application of oxygen, dobutamine and furosemide followed by angiotensin converting enzyme inhibition and digitalis. In the echocardiographic control examination 14 days later left ventricular function had normalized. The changes of electrocardiogram normalized 4 weeks later.

Aged↗
PubMed1990

The effect of sustained release application of chlorhexidine on salivary levels of Streptococcus mutans in partial denture wearers.

The effect of a slow-releasing dosage (SRD) coating of chlorhexidine on the salivary levels of Streptococcus mutans and on plaque index scores in patients with removable partial dentures (RPD) was tested. The SRD proved to be effective in maintaining a low level of S. mutans counts after mechanical cleaning, as compared to a baseline established during the control period. Plaque index scores were lower following the treatment and correlated with the microbiological results. Our findings indicate that a single application of sustained-release chlorhexidine to removable partial dentures effectively maintains S. mutans levels as well as reducing the plaque score for a minimum period of 1 week.

Chlorhexidine↗
PubMedDate not supplied

[A method for the quantitative determination of reticulocytes in induced reticulocytosis].

A technique accelerating and simplifying the determination of reticulocyte counts in animals with reticulocytosis has been suggested. The results are correctly evaluated without microscopy. The technique can be also used for the identification of patients with hemolytic anemias, especially in field conditions. An application for the technique developed has been submitted and positive approval has been obtained on November 20, 1987.

Animals↗
PubMed2000

A simplified application of systematic field sampling and low-cost video recording set-up for viewing disector pairs - exemplified in the rat cochlear nucleus.

A description of a simple and efficient way of systematic field sampling along with a low-cost set-up for simultaneous viewing of physical disector pairs at the light microscopic level is presented. So far the use of a programmable motorized stage for the former purpose and two projection microscopes for the latter have proved to be the most efficient and fastest solutions, but the relatively high expense of purchasing such equipment constitutes a major impediment to the widespread use of such stereological methods. We describe practical applications of two new cost-effective alternative approaches derived from existing ones. These approaches are used in estimating the total number of neurones in the cochlear nucleus of the rat and involve the systematic selection of section fields at a magnification significantly lower than the magnification used for counting and a video recording set-up for comparing section pairs. Apart from providing a cheap way for the implementation of the physical disector, the main feature of the application, which may also encourage its use, is that it provides an efficient and simple design for systematic area sampling without requiring any specialized equipment. The same is true for optical disector designs, where the procedure is simplified even further.

Animals↗
PubMed2001

Semiquantitative analysis of SPECT images using 99Tc(m) -HMPAO in the treatment of brain perfusion after the attenuation correction by the Chang method and the application of the Butterworth filter.

The aim of this work was to optimize the parameters for semiquantitative analysis with regard to the average number of counts per pixel in tomographic projections. The analysed studies were divided into three groups, proportionally, to average the counts per pixel. The analysis of results proved the dependence (P<0.05) between the average geometric contrast of images with the maximum counts per pixel higher than 500, reconstructed with the cut-off frequency fc=0.50fn, and the standard image. Also, the same dependence was found between the group with an average of 200 counts per pixel and the standard image at the cut-off frequency fc=0.40fn. The analysis of the attenuation coefficient, c, showed significant differences. The attenuation coefficient c=0.12 cm-1 as accepted for further studies. Optimization of the matrix order of interpolation filter proved increasing of contrast with decreasing size of the matrix. In conclusion, according to the average number of counts per pixel in a projection the parameters of image reconstruction, particularly the cut-off frequency fc of the Butterworth filter, should be changed. Additionally, an attenuation correction with coefficient c=0.12 cm-1 and interpolation with matrix size equal 7x7 should be applied.

Brain↗
PubMed1970

Evaluation of ultra-low-volume insecticide dispensing systems for use in single-engined aircraft and their effectiveness against Aedes aegypti populations in South-East Asia.

An evaluation study of ultra-low-volume (ULV) spraying of insecticide from aircraft was carried out in Thailand, to determine if this technique could be used for the emergency control of Aedes aegypti, the major vector of haemorrhagic fever. A small, single-engined aircraft, a Cessna-180, was used in the trials and 2 types of spraying equipment were tested; both were found to be equally effective. The aircraft was fitted with 6 spraying nozzles and flew at an altitude of 150 feet (46 m) at a speed of 100 miles/h (161 km/h). The insecticide used was 95% technical grade malathion and swaths 75 feet wide (22.8 m) were laid down; the rate of application was 3 US fl oz/acre (219 ml/ha).Trials were made in 3 villages near Bangkok and it became apparent that a small aircraft could not produce the required even distribution of insecticide; the rate of application was therefore increased to 6 US fl oz/acre (438 ml/ha). This increased rate appeared to compensate for the narrow width of the swath and produced very satisfactory mortalities in caged mosquitos as well as in natural populations.The size and distribution of droplets was monitored by the use of oil-sensitive red dye cards which showed that there was a good penetration of insecticide into dwellings, etc. Trial results were evaluated by biting counts, bioassays of Aedes and Culex adults and larvae, Culex dips and ovitraps. Biossays indicated that the 6 US fl oz/acre rate of application was almost 100% effective in the open and produced satisfactory mortalities inside markets and dwellings.It was concluded that larger aircraft would be required to treat areas of more than 1000 acres (405 ha) and congested city areas but that the rate of application of insecticide could be considerably lower. Nevertheless, small aircraft can be useful in smaller and less congested areas.

Aedes↗
PubMed1978

[Experimental investigations on the regeneration of aortic endothelial cells. Automatic and visual evaluation of autoradiograms (author's transl)].

Autoradiographic studies on plane preparations using 3H-thymidine and pulse cytophotometric investigations on scrape preparations of aortic endothelium from 34 rabbits have brought the following results: By means of multiple 3H-thymidine-injections the number of labelled endothelial cells is higher in young growing animals than in older rabbits; pulse cytophotometrically we did not find any indication of polyploidization processes; from that we conclude a decrease of the growth fraction with advancing age. In comparison to visual counting the automatic image analysis is a highly effective method for the evaluation of autoradiograms. After multiple 3H-thymidine applications the mean blacking intensity of the labelled cell nuclei is lower in 2 months old animals than in 12 months old rabbits; that may mean an increase in the S-phase duration of the proliferating endothelial cells of the older animals. In the area with branching sites there are more 3H-thymidine-labelled endothelial cells than in the neighbouring vascular segments. The significance of these findings for the preferential incidence of arteriosclerosis in this area is discussed.

Age Factors↗