[Cognitive impairments and microvascular endothelial dysfunction in unilateral occlusion of the carotid artery]. / Kognitivnye narusheniia i disfunktsiia éndoteliia mikrososudov pri unilateral'noi okkliuzii sonnoi arterii.

OBJECTIVE: The study was aimed at determining cognitive impairments, vascular endothelial dysfunction and morphofunctional alterations of neurons in brain structures (neocortex and hippocampus) in unilateral occlusion of the common carotid artery in albino rats. MATERIAL AND METHODS: The experiments were carried out on Wistar albino rats. The animals were divided into two groups: group 1 - the control group composed of sham-operated rats and group 2 - the study group consisting of rats with cerebral ischaemia. The model of cerebral ischaemia was reproduced by occlusion of the left common carotid artery. Cognitive functions in rats were assessed by means of the object recognition test and conditioned passive avoidance response (CPAR). The degree of vascular endothelial dysfunction was evaluated by the number of circulating endotheliocytes and concentration of end products of nitric oxide - nitrites in blood plasma. Pathomorphological studies of the brain and morphometric analysis of the number of damaged neurons after ischaemic exposure were carried out by means of light microscopy. RESULTS: It was determined that unilateral occlusion of the common carotid artery in rats resulted in memory impairment revealed by the object recognition test and CPAR (p≤0.05). Cerebral ischaemia induced an elevated level of circulating endotheliocytes and a decrease in end products of nitric oxide - nitrates as compared with the controls (p≤0.05). Morphological study demonstrated signs of vasoconstrictive reactions, microvascular endothelial oedema, as well as an increase in the proportion of damaged neurons localized in the fronto-bregmatic region of the cortex of the cerebral hemispheres and hippocampus on the ipsilateral side (p≤0.05). CONCLUSION: Unilateral occlusion of the common carotid artery in albino rats resulted in cognitive impairments, damage of neurons in the most vulnerable areas of the cortex of the cerebral hemispheres and hippocampus predominantly on the ipsilateral side. Cognitive impairments and ischaemic lesions of the brain structures are induced by endothelial dysfunction, enhanced desquamation of endotheliocytes and prevalence of vasoconstrictive reactions resulting from decreased production of the major vasorelaxing factor - nitric oxide.

[Atypical maxillofacial shot wound]. / Atypické strelné poranenie maxilofaciálnej oblasti.

Maxillofacial shot wounds belong to the relatively less frequent injuries. The soft tissues might be affected in particular, however the most of injuries are combined with infliction of hard tissues. Shot wounds by the bow or crossbow are very rare. They might result as a consequence of either accident or suicide. In the literature some cases of a suicidal experiments had been published, all of these cases were due to arrow from the crossbow. There was no injury being documented due to arrow from the bow. The authors present a case of maxillofacial injury done by shoot of arrow from the bow in the course of a child's play.

[Scintigraphic detection of osteoblast activity after implantation of BAS-0 bioactive glass-ceramic material into long bone defects]. / Scintigrafické vysetrení osteoblastické aktivity po implantaci bioaktivního sklokeramického materiálu BAS-0 do defektu dlouhých kostí.

PURPOSE OF THE STUDY: The aim of the study was to demonstrate, by three-phase bone scintigraphy, radionuclide uptake at the site of defects in long bones filled with the non-resorbable bioactive glass-ceramic material BAS-0 at a long follow-up. MATERIAL: Twenty patients, 14 men and 6 women, operated on between 1990 and 2000 for benign bone tumors or tumor-like lesions localized in the femur, tibia or humerus were comprised in the study. Their average age at the time of operation was 14 years (range, 8 to 24). The diagnoses based on histological examination included juvenile bone cysts in 11, aneurysmal bone cyst in five, non-ossifying fibroma in two, and fibrous dysplasia in two patients. The lesions were localized in the femur, humerus and tibia in 11, five and four patients, respectively. The metaphysis was affected in eight and the diaphysis in 12 patients. METHODS: Clinical, radiological and scintigraphic examinations were carried out at 2 to 12 years (7 years on average) after surgery. The clinical evaluation included subjective complaints and objective findings. Radiographs were made in standard projections and the osteo-integration of glass-ceramic material was investigated. Three-phase bone scans were made and the healthy and the affected limbs in each patient were compared by means of an index. Radionuclide uptake was considered normal when the index value was equal to 1.0, mildly increased at an index value of 1.2, moderately increased at 1.2-1.5 and markedly increased at an index value higher than 1.5. RESULTS: The clinical evaluation showed that, in the patients with glass-ceramic filling of metaphyses, six had no subjective complaints and two reported transient pain. In the patients with implants in diaphyses, subjective complaints were recorded in nine and no complaints in three patients. No inflammatory changes in soft tissues were found. No restriction in weightbearing of the limb treated was reported by any of the patients. On radiography, 18 patients were free from any disease residue or recurrence. Two patients had a residual defect. The bioactive glass-ceramic material BAS-0 was completely incorporated in all patients. On three-phase bone scans, radionuclide distribution on the flow phase and soft tissue phase was symmetrical in both limbs of all patients. For the metaphyseal location of implants, the delayed images demonstrated physiological radionuclide distribution in one patient, mildly increased distribution (index up to 1.2) in four, increased uptake (index up to 1.5) in two patients, and highly increased uptake (index above 1.5) in one patient. For the diaphyseal location of implants, the delayed scans demonstrated slightly increased radionuclide distribution in two, markedly increased in two and highly increased uptake in eight patients. DISCUSSION: The tissue during incorporation of a non-resorbable synthetic material is influenced by stress-shielding. This changes local mechanical signals, which has a negative effect on the adjacent bone tissue. Stress accumulating at the interface of a rigid implant and bone tissue may result in pain, and is detected by scintigraphy as an increased nucleotide uptake, particularly in diaphyseal grafts. CONCLUSIONS: This paper presents problems associated with implantation of the non-resorbable bioactive glass-ceramic material BAS-0 in the treatment of diaphyseal defects of long bones. The results show that, for filling of the defects described herein, non-resorbable glass-ceramic materials are not suitable.

99mTc-aminohexylidendiphosphonate and 99mTc-pyrophosphate in the scintigraphic diagnosis of experimental myocardial infarction in dogs.

99mTc-aminohexylidendiphosphonate (99mTc-AHDP) is a new Czechoslovak pharmaceutical of the phosphonate line which contains the amino group NH2 in its molecule. This substance was tested in 5 dogs with experimentally provoked 48-h old myocardial infarction. The in-vivo scan and the radioactivity of tissue samples demonstrated that 99mTc-AHDP is as suitable for imaging acute myocardial infarction as is the commonly used 99mTc-pyrophosphate.

Selective medium for primary isolation of members of the tribe Proteeae.

A selective Proteeae medium (SPM) for isolation and preliminary detection of species of genera Proteus, Morganella, and Providencia was evaluated. The SPM contains tryptose phosphate agar with phenolphthalein monophosphate (as substrate for phosphatase activity), bile salts and polymyxin B (as inhibitors). The selectivity of the SPM was tested by the ecometric method of quality assurance of culture media. Fourteen reference cultures of enterobacteria and fifty-four strains of Proteeae were tested for their absolute growth index (AGI). Ninety-five percent of tested Proteeae strains display an AGI above 2.5. The detected phosphatase activity proved to be able to discriminate colonies of members of the tribe Proteeae. The ability of SPM for primary isolation of members of Proteeae was tested on food and clinical material and 94 strains were isolated. In addition, the SPM was employed in routine practice of clinical microbiology. From 1016 clinical samples (stool, urine, vaginal and urethral swabs), 57 strains of Proteeae were detected by the SPM in contrast to 35 strains by the routine procedure. The difference amounts to nearly 40%.

[A rapid method of detecting psychrotrophic bacteria in raw milk]. / Zrychlená metoda stanovení psychrotrofních baktérií v syrovém mléce.

Determination of the counts of psychrotrophic bacteria offers an important indicator of the technological quality of purchased raw milk, and also an indicator of the hygienic level of its production in agricultural primary production, particularly with respect to the sanitation of milking machines. The applied and published methods of determination of the counts of psychrotrophic microbial contaminants have revealed large differences, multiplied by insufficient legislation in this country, by vague and even wrong definition of the given group of bacteria, and/or by the misuse of the concepts psychrotrophic and psychrophil bacteria. Since the level of the microbial quality of raw milk should be increased, we investigated and developed convenient methods of determination of psychrotrophic bacterium counts in the last years (Urbanová et al., 1988; Urbanová, in print): these methods should facilitate the reasonable control applying an expeditious procedure. A long incubation time of standard procedure is a disadvantage complicating the practical use of the previous procedure (10 days at a temperature of 7 degrees C). An accelerated procedure of psychotrophic bacterium cultivation to determine their counts in raw milk is described in this paper. After thorough stirring, a raw milk sample is diluted with sterile physiological solution (pH 7.2) pursuant to the standard CSN 57 0101. The dilutions 10(-2), 10(-3), 10(-4) were the most convenient for inoculation when the counts of psychrotrophic contaminants were determined in raw milk. The dilutions 10(-3), 10(-4), 10(-5) were inoculated if higher milk contaminations were expected.(ABSTRACT TRUNCATED AT 250 WORDS)

[Identification of Citrobacter species and their occurrence in raw products and foods]. / Identifikace druhu rodu Citrobacter a jejich výskyt v surovinách a potravinách.

Fifty-nine strains of bacteria were tested in study that were isolated from raw materials and foods (raw milk, farm milking parlors, sausage meat, raw potatoes, cheese, frozen oven-ready foods, confectionery products, cold dishes), and they were included in the genus Citrobacter using a commercial diagnostic kit ENTEROtest 16 (Lachema a.s., Brno), numeric identification program TNW (Czech Collection of Microorganisms, Faculty of Science of Masaryk University at Brno) and identification key (O'Hara et al., 1995). The results of the ENTEROtest itself, including OXI and ONPtests, did not provide satisfactory discrimination of detected strains to the level of species since 64.4% were identified by the genus by TNW program or designated as intermediary strains. Correct species identification was obtained in 33.9% only (Tab. I). Five next conventional tests (dulcitol, alpha-methyl-glucoside, raffinose, melibiose, arginine dihydrolase) were used and their results successfully specified 94.9% tested strains to the species level (Tab. I). A dichotomic identification key (O'Hara et al., 1995) enabled to classify the strains on the basis of the results of ENTEROtest 16 and two conventional tests (dulcitol, melibiosis) relatively easily, and it can be recommended for routine identification of typical Citrobacters from foods. Only ten strains were classified in a wrong way (16.9%) due to false results in ENTEROtest (Tab. II). Tab. III shows the strains classified wrongly by both identifications systems. The tested set contained Citrobacter braakii in a majority of cases (30 strains), followed by C. freundii (17 strains), C. youngae (6 strains), 2 strains of genomospecies 10 and one strain of C. koseri, C. amalonaticus and C farmeri. Tab. IV shows the presence of Citrobacter species in the particular raw materials and foods. The most frequently present species in raw materials: C. braakii (26 strains, 68.4%), C. freundii (3 strains, 7.9%) and C. youngae (3 strains, 7.9%). Foods contained these three species only: C. freundii (14 strains, 66.7%), C. braakii (4 strains, 19.0%) and C. youngae (3 strains, 14.3%). The percentage of the three most frequently present species in raw materials is substantially different from their percentage in foods (Fig. 1). The higher percentage of C. freundii presence in foods can be ascribed to secondary contamination.

[A computerized numerical identification system for the diagnosis of bacteria isolated from raw animal food products]. / Pocítacové numerické identifikacní systémy v diagnostice baktérií izolovaných z zivocisných surovin.

Acceleration of bacteria identification and certain standardization of results in clinical, and also veterinary and food microbiology is enabled by use of commercial diagnostic sets and computer processing of their results. In the present paper, 80 bacterial strains isolated from animal sources in the department of food hygiene were used for assessment of two numerical identification systems (TNW and IDENTI). They were isolated from primary cultures on End's, meat-peptone and blood agar (Imuna, HI-Media) under different culture regimes (37 degrees C/24 h, laboratory temperature/3 days, 7 degrees C/10 days). A commercial diagnostic set ENTEROtest 1 and 2 (Lachema a.s., Brno, Czech Republic) was used for the purposes of diagnostics of gram-negative fermenting, oxidase-negative bacteria. Testing was performed according to the manufacturer's instructions by the culture of isolated strain. The test results were read continually after 6-hour (sugars) and 24-hour cultivation. Each strain was tested three times. Additional tests--oxidase, catalase, yellow pigment production. These commercial numerical identification systems were compared: IDENTI (supplied by Z. Svoboda, Jihlava; Lachema a.s., Brno) TNW (supplied by the Czech Collection of Microorganisms, Brno) Out of the total number of 80 isolated gram-negative bacteria 77 (96.25%) strains were, classified into the genera by both numerical systems. This result is in agreement with the percentage of identified strains by means of foreign commercial sets and numerical systems, which ranges from 93 to 99% in bacteria isolated from foods (Cox and Mercuri, 1979).(ABSTRACT TRUNCATED AT 250 WORDS)

[The NEFERM test in the identification of psychrotrophic bacteria isolated from milk and dairy products]. / Uplatnení NEFERM-testu v identifikaci psychrotrofních bakterií izolovaných z mléka a mlécných výrobku.

Sanitary and nutriti on quality of foods from animal sources, that means also of milk and dairy products, are significantly influenced by the presence of psychrotrophic bacteria, the lipolytic and proteolytic activity of which contributes to their nutritive and sensory changes. This applies particularly to psychrotrophic bacteria of the genus Pseudomonas, Shewanella, Alcaligenes and Flavobacterium (Muir and Phillips, 1984; Piton and Richard, 1985; Craven and Macauley, 1990). These bacteria cause raw and pasteurized milk to turn bitter and gelatinize, and/or technological problems during processing. In the present paper the possible use of NEFERM-test and numerical identification system TNW was verified for identification psychrotrophic, nonfermentative bacteria isolated from milk and dairy products. This research is a continuation of our preceding work (Urbanová and Pácová, 1995) dealing with identification of these bacteria in poultry. The test comprised 35 psychrotrophic, gram-negative, oxidase-positive nonfermentative bacteria isolated from raw milk (30 strains) and hard cheeses (5 strains). A commercial diagnostic kit NEFERM-test (Lachema a.s., Brno) and respective conventional methods were parallelly used for identification: the conventional methods were complemented by assays of oxidase (OXI) and beta-galactosidase (ONPG) using of commercial strips Lachema a.s. Additional tests were as follows: gelatin and Tween 80 hydrolysis, fluorescein production. The readings of yellow or orange pigment production were also applied. A numerical identification system TNW was used to process the results (Czech Collection of Microorganisms, Faculty of Science, Masaryk University, Brno). Correct identification of bacteria by means of commercial diagnostic kits depends particularly upon the high goodness of fit of the results obtained by conventional tests in relation to those obtained by commercial kits.(ABSTRACT TRUNCATED AT 250 WORDS)

[The NEFERM-Test for identification of psychrotrophic bacteria in food]. / Uplatnení NEFERM-testu v identifikaci psychrotrofních bakterií izolovaných z potravin.

Identification of gramnegative nonfermentative bacteria by traditional methods requires much labor and time consumption. Therefore in this study we tested the use of an available commercial diagnostic kit and processing of its results by computerized numerical identification systems. Identification involved 83 gramnegative, psychotrophic, nonfermentative, oxidase-positive bacteria isolated from just slaughtered and deep-frozen chickens. The strains were isolated from primary cultures in Psedomonas F and Endo agar, Violet Red Bile agar (IMUNA, OXOID) and King B agar (King et al., 1954), under two cultivation regimes (room temperature for three days and 7 degrees C for 10 days). A commercial kit for diagnostics of nonfermentative bacteria in clinical microbiology under the name NEFERM-test (Lachema a.s., Brno) containing 12 biochemical assays was used for strain identification: GLU, AGR, IND, ESL, SCI, PHS, URE, MAN, XYL, MLT, LAC, NIT. All these assays were conducted parallelly by traditional methods. Additional tests involved the use of OXI and ONPG commercial strips (Lachema a.s., Brno), gelatin and Tween 80 hydrolysis (Pácová and Kocur, 1984) and fluorescein production (King et al., 1954). These identification systems were used to process the results of tests; index and differentiation table (Lachema a.s., Brno) and numerical computerized systems TNW (Czech Collection (Z. Svoboda, Jihlava). Various shortened procedures, identification keys and systems are used to speed up identification of gramnegative nonfermentative bacteria. Available commercial identification kits (API NE20) in form of microassay have been developed particularly for diagnostics of bacteria from clinica materials, which applies to NEFERM-test of the Czech make.(ABSTRACT TRUNCATED AT 250 WORDS)

[Determination of the number of somatic cells in milk using the rapid diphenylamine DNA filter method]. / Stanovení poctu somatických bunek v mléce rychlou difenylamin DNA filtr metodou.

The rapid reaction of the diphenylamine agent with DNA was used for the determination of the counts of somatic cells in cow's milk, using the DNA filter method. The method is based on the filtration of a warmed (65-70 degrees C) mixture of milk with Triton X-100 through the Synpor nitrocellulose membrane filter, pore size 2 to 5 microns, and subsequent DNA determination of the collected somatic cells by the colour reaction of diphenylamine. A 2ml quantity of distilled water and 4 ml of diphenylamine reagent were added to the membrane filters with somatic cells. The mixture is warmed in water bath at 90 to 100 degrees C for 20 min., then it is cooled, centrifuged (3500 X g, 15 min.), and the optical density is measured at 595 nm. The relation 8 micrograms = 1 million cells was used for the conversion of DNA content to the counts of cells. The average variation coefficient of the determination was 5.9% and the coefficient of correlation between the diphenylamine DNA filter method and the direct microscopy of the somatic cells on membrane filters was r = 0.997. Using the diphenylamine DNA filter method, the counts of somatic cells can also be determined from milk samples stored in frozen condition or from the filters with collected cells kept at the temperature of 4 degrees C (10 days) or 25 degrees C (3 days). Milk stabilized with formaldehyde can also be used for the determination if stored at 4 degrees C.

[Methods of diagnosing subclinical mastitis in dairy cow mastitis control programs]. / Metody diagnostiky subklinických mastitid v programu tlumení mastitid dojnic.

The number of somatic cells and the isolation of the causative agents of mastitis in quarter, composite, bucket, and bulk tank samples of cow's milk was determined four times during a six-month period. The number of somatic cells in milk samples indicated a degree of mastitis infection and was influenced neither by the year season nor by the length of lactation. At a repeated examination of 28 dairy cows an increased number of somatic cells in milk was found once in 68 udder quarters and with three successive samplings only in 21 quarters. The etiological agents of mastitis were detected once in 31 quarters and three times in succession only in five quarters. The number of cows positive by the number of cells in quarter samples of milk increased from 52.9-58.8% at a single examination to as much as 100% at four examinations. The etiological agents of mastitis were isolated in a single examination in 17.6% of cows and at four examinations in 58.8% of cows. The composite and bucket samples of milk containing 200 to 300 thousand cells per ml are recommended to be considered as mastitis-positive: in 68 to 78% they came from cows having more than 500 thousand cells per ml at least in one quarter sample. The number of cells in a bulk sample was in correlation with the percentage of cows having a positive NK-test (similar to CMT) and positive isolation of S. agalactiae from quarter milk samples.(ABSTRACT TRUNCATED AT 250 WORDS)

[Use of the Synpor membrane filter for the separation and determination of the number of somatic cells in the milk of dairy cows using the indole DNA filtration method]. / Vyuzití membránových filtru Synpor pro separaci a stanovení poctu somatických bunek mléka dojnic indol DNA filtr metodou.

The somatic cells of cow's milk were separated by filtering through nitrocellulose membrane filters (Synpor), pore size 2 to 5 micron. An indole reagent was added to the membrane filters with trapped cells, the mixture was warmed in boiling water bath for 20 min., then cooled, centrifuged (3500 X g, 15 min.), and optical density was measured at 490 nm. The relation 8 micrograms = 1 million cells was used for the conversion of DNA content in sample to the counts of cells. The average variation coefficient of the determination was 4.7% and the coefficient of correlation between the indole DNA filter method and direct microscopy on membrane filters was r = 0.997. Using the indole DNA filter method, the counts of somatic cells can be estimated from milk samples kept frozen or from filters with retained cells kept at 4 degrees C, or (for a short time--3 days) at 25 degrees C. The estimation is not distorted when the milk samples are stabilized with K2Cr2O7 or a mixture of K2Cr2O7 and HgCl2, if they are kept at 4 degrees C for eight days.

[Incidence of foreign substances in raw and pasteurized milk]. / Výskyt cizorodých látek v syrovém a pasterovaném mléce.

In 1980 to 1982, the occurrence of inhibiting substances, herbicides and chemical elements contaminating raw farm and bulk milk and pasteurized milk in different stages of technological processing was studied in the input regions of 19 dairies of the Lacrum Brno concern corporation. Residues of inhibiting substances were demonstrated by the thin-layer microbiological diffusion disk method using the testing strain of B. stearothermophilus var. calidolactis, to be present in raw milk (9.1% out of 143 samples) as well as in pasteurized milk (2.0% out of 151 samples). Chromatographic method (sensitivity 0.01-0.05 mg.kg-1 milk) was used for examining 135 raw milk samples for the occurrence of herbicides based on diazines and triazines. Herbicides were demonstrated to get into milk; residues were found in 45 samples (33.3%). Atomic absorption spectrophotometry was used for examining 143 raw milk samples for the contents of zinc, copper, iron and manganese. As to zinc content, 100% of the samples met the standard, as to iron content 99%, and as to the content of copper, 89% (Ministry of Health Directives, No. 50, 1978). The content of manganese remained within the range up to 0.1 mg.kg-1 milk in 60% of the samples. Out of the 135 milk samples (101 raw, 34 pasteurized milk ones) examined for the content of lead, four samples (3%) failed to meet the limit of the highest admissible level.

[Monitoring the incorporation of bioactive glass ceramics in the area of the acetabulum after surgical revision of the hip joint arthroplasty by scintigraphy.]. / Sledování postupu vhojování bioaktivní sklokeramiky v okolí acetabula po revizní operaci endoprotézy kycle pomocí scintigrafie.

The authors investigated changes of osteoblastic activity at the site of a reconstructed defect of the acetabulum after aseptic loosening of a total endoprosthesis of the hip joint by administration of an osteotropic radiopharmaceutical preparation, methylendiphosphonate, labelled with 99mTc. The defect of the acetabulum was filled with bioactive glass ceramic material BAS-0 (Lasak Co., Prague) in the shape of dense granules 4-8 mm in size and a metal antiprotrusion basket. Into the reconstructed acetabulum a new polyethylene socket of the endoprosthesis was implanted. Examination by the two-head camera Helix of Elscint Co. was performed within two weeks after operation. Healing in the described group of ten patients was without complications, after three months the patients burdened the operated joint partly, after six months they burdened it by half their weight and after another three months by their whole weight. Immediately after operation the metabolic acitivity in the vicinity of the defect exceded the reference activity of normal bone by 50% The authors discuss possibilities and advantages of the mentioned examination in investigations of the incorporation and function of glass ceramic prostheses of osseous tissue. Key words: bioactive glass ceramics, scintigraphy, osteotropic radiopharmaceutical preparation, 99mTc methylendiphosphonate, reconstruction of acetabular defect.

[Cysts and pseudocysts of the spleen in children]. / Cysty a pseudocysty sleziny u detí.

A cyst in the upper pole of the spleen was diagnosed in three children. In all the pole of the spleen was resected -in two instances it contained a cyst, in one a pseudocyst. The postoperative course was without complications. All patients had scintigraphic check-up examinations, in all three instances functional lienal tissue was found which did not differ much as to its size from that of a normal spleen. In cysts situated in the splenic pole the authors consider partial resection of the affected portion of the spleen as the optimal surgical method. In cysts of other portions of the spleen it would be better to select a different surgical procedure, e.g. enucleation of the cysts, partial decapsulation of the cyst or percutaneous puncture of the cyst combined, if necessary, with sclerotization.