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[The importance of polymerase chain reaction in the diagnosis of enterovirus infections of the central nervous system in children. Clinico-epidemiologic characteristics].

BACKGROUND: To compare the diagnosis utility of a reverse transcription-polymerase chain reaction (RT-PCR) for detection of enteroviral RNA in cerebrospinal fluid (CSF) in comparison to viral culture for central nervous system infections in pediatric population and to know the clinic epidemiological characteristics of this infection. METHODS: From June to December of 1997, 116 CSF samples of children were included in the study. The samples were inoculated in MRC-5 and HEP-2 and the enterovirus RNA was detected with AMPLICOR-Enterovirus (Roche Diagnostic System). The virus were typed by neutralization. RESULTS: 36 samples were positive (30 were positive for RT-PCR and 6 for RT-PCR and viral culture). The media time in detect the CPE was 4.8 days. The viruses found were four echovirus 6, one echovirus 30 and one echovirus 7. Fifteen children were under 6 months (93.3% under two months) and 21 children over 6 months with a media age of 81 months (range, 38-160 months). In over 6 months old children, the most frequent clinical presentation was meningitis aseptic syndrome with pleocytosis and in under 6 months old ones was fever and only 60% of these children had pleocytosis. The evolution was good in all the cases. CONCLUSIONS: AMPLICOR-Enterovirus is a sensitive method for the diagnosis of enteroviral meningitis. The clinical manifestations are different with the age and due to the absence of neurological symptoms and pleocytosis in the patients under two months, we think that it is necessary to study the presence of enterovirus with genetic amplification methods in this population of patients.

Central Nervous System Infections↗

Implantable insulin pumps: infections most likely due to seeding from skin flora determine severe outcomes of pump-pocket seromas.

Complications at implantation site of implantable insulin pumps may lead to premature removal. To elucidate the origins and the outcomes of these local adverse events. We investigated seromas of the 'pump-pocket' that have been detected for an eight month-period during the follow-up of such-treated forty type 1 diabetic patients. At the start of study period, skin bacterial flora was sampled at umbilicus and groin, and isolated strains of Staphylococcus epidermidis were preserved in specific vials at -20 degrees C. Each time a seroma was detected at transcutaneous 45 days-refill of pump reservoir, it was sampled for bacterial cultures. Isolated strains of S. epidermidis from seroma were genetically compared to preserved strains of corresponding patients using Pulsed-Field Gel Electrophoresis (PFGE) after genomic restriction by SmaI. Among the ten seromas that occurred after a mean time of 9.9 months since implantation, S. epidermidis were isolated in five cases. Genetic comparison of isolated strains could be performed in three cases. Compared strains showed identical (in 2 cases) or closely related (in one case) PFGE profiles. While the five aseptic seromas resolved with rest, four infected cases required explantations after one to nineteen months in spite of antibiotic therapy and the fifth one persisted without impairment under long-term antibiotics. Our results suggest that seeding from the skin flora is a key-factor determining the severity of pump-pocket complications. We recommend that bacterial investigations of pump-pocket seromas should be systematically performed, while prophylactic measures might include antibiotic cover for each puncture of the pump-pocket.

Adult↗

[Technics of preparation and collection of embryonal nerve tissue for transplantation in clinical practice].

The proper author's experience is used in discussing possibilities on how to obtain the embryonal nerve tissue's samples for transplantation in various CNS diseases. The most reliable, though a unique one is the obtention of the whole foetus. This may be exclusively done by the miniabortion using suction curettage, or eventually at greater dilatation of cervix with the use of fine abortive forceps providing the ultrasound control. The most secure source of tissue in order to make transplantation is the removal of pregnant uterus, e. g. for its myomatosis. The proper preparation of tissue for transplantation is provided with maximally aseptic conditions.

Abortion, Induced↗

Response to alpha-interferon in children with Philadelphia chromosome-positive chronic myelocytic leukemia.

The therapeutic efficacy and toxicity of alpha-interferon (alpha-IFN) (Roferon, Hoffmann-La Roche, Inc., Nutley, NJ) were determined in 15 children (age range, 6 to 20 years) with Philadelphia chromosome-positive chronic myelocytic leukemia (Ph+ CML). All patients had received cytoreductive therapy with either hydroxyurea (n = 13) or busulfan (n = 1) or both (n = 1) for 6 weeks to 46 months (median, 7 months) before beginning alpha-IFN therapy at a dose of 5 x 10(6) U/m2/d intramuscularly. This dose was escalated to 10 x 10(6) U/m2/d if leukemia was inadequately controlled. Ten children had a hematologic response, with nine showing a reduction in the percentage of Ph+ marrow cells, including four who had no detectable Ph+ cells in marrow samples collected 48 to 204 weeks after the initiation of therapy. Two of 15 patients remain free of Ph+ cells. Therapy was discontinued before week 104 in ten patients because of the following: (1) early hematologic responses without a decrease in Ph+ cells (three patients); (2) early resistant disease (one patient); (3) blast crisis (one patient); (4) progressive disease (two patients); (5) seizure attributed to high-dose alpha-IFN (one patient); or (6) an inadequate trial of alpha-IFN caused by aseptic necrosis or poor compliance (two patients). The most common side effects were mild and have included fever, malaise, headache, myalgias, and pain at the injection site. Adverse events causing interruption of therapy were seizures, aseptic necrosis, and myelofibrosis. alpha-IFN stabilizes the chronic phase of Ph+ CML in some children, is adequately tolerated when administered at a dose of 2.5 to 5 x 10(6) U/m2/d intramuscularly, and results in a significant decrease in the proportion of Ph+ metaphases in some patients. alpha-IFN in combination with an effective cytoreductive agent or agents appears worthy of further clinical testing in this disease.

Adolescent↗

Detection of epithelial cell transfer in spinal areas by light microscopy and determining any tissue coring via cell culture during combined spinal-epidural interventions.

BACKGROUND AND OBJECTIVES: Epithelial tissue coring by spinal needles during subarachnoid injections may cause intraspinal epidermal tumors. Previous studies have investigated tissue transfer with different needle types during subarachnoid or epidural injection. This study deals with the transfer of epithelial tissue during combined spinal-epidural (CSE) anesthesia. METHODS: We studied 68 American Society of Anesthesiologists I to III adult patients. CSE anesthesia was induced under aseptic conditions at the L2-3 or L3-4 interspace with patients in the lateral decubitus position. Cerebral spinal fluid, spinal needle stylet, fluid used to flush the interior of the spinal needle, fluid used to wash the exterior of the spinal needle, fluid used to flush the interior of the epidural needle, and fluid used to wash the exterior tip of the epidural needle were examined under light microscopy (n = 30 patients) or incubated in a cell-culture medium (n = 38 patients). Samples were incubated in cell-culture medium alone (n = 13) or in a cell-culture medium for 3 weeks and then in a medium with epidermal growth factor (n = 25). As a positive control, skin tissue samples were taken by punch biopsy from 10 randomly chosen patients who underwent CSE interventions. These samples were incubated in an enriched medium serum. RESULTS: Light microscopy revealed that there was cell transfer in all phases in various rates: samples 1, 2, 3, 4, 5, and 6 contained epithelial cells and debris in ratios of 6.9%, 20.7%, 6.9%, 20.7%, 26.7%, and 33.3%, respectively. Epithelial cell colonization was detected in the cell-culture samples taken from the control group but not in the samples taken from the CSE group. CONCLUSIONS: We could not reproduce the cells or cell debris obtained during the CSE interventions in vivo, which can be explained by a possible structural deformation of cells or the inadequacy of the amount of cells that were transferred.

Adult↗

An outbreak of viral meningitis associated with a public swimming pond.

From July to October 2001, 215 cases of aseptic meningitis occurred among the inhabitants of the German city of Kassel and neighbouring counties. A matched case-control study identified bathing in a public, nature-like pond during the beginning of the outbreak as a risk factor for disease [matched odds ratio (mOR) 44.8, 95% confidence interval (CI) 3.9-515.6]. Among bathers, patients with meningitis spent more time in the water (mOR 18.8, 95% CI 2.0-174.1) and swallowed water more frequently (mOR = 7.3, 95% CI 0.7-81.8). Of 30 cerebrospinal fluid samples tested, echovirus 30 was cultured from 16, and echovirus 13 from seven. An echovirus 30 sequence obtained from one pond water sample showed a 99% nucleotide and 100% amino-acid homology with patient isolates. This outbreak demonstrates the potential of nature-like swimming ponds to cause widespread community infection with substantial public health impact.

Adolescent↗

In vitro propagation of Withania somnifera and isolation of withanolides with immunosuppressive activity.

Withania somnifera plantlets were produced in vitro from the shoot-tip of aseptically germinated seedlings. Culture conditions were optimized using different plant growth regulators which gave rise to 120 shoots from a single bud. The plantlets were then transferred to pots and maintained in greenhouse for 4 months. 90% of these in vitro propagated plantlets survived and showed normal growth. Leaves from these plants were used for isolation of the withanolides. Methanolic extract of leaves from plantlets growing in tissue culture and those transferred to the greenhouse were evaluated for immunomodulatory activity. While the extract from greenhouse samples showed potent immunosuppressive activity, those from tissue cultures samples did not show any activity. Fractionation and characterization of withanolides, using HPLC, NMR, MS methods revealed the presence of withaferin A in the greenhouse samples. Our results indicate that Withania species may require longer time and better differentiation and also natural environment for the production of withaferin A.

Adjuvants, Immunologic↗

Induction of hydrometra in goats by means of active immunization against prostaglandin F2 alpha.

Hydrometra is a pathological condition of the uterus which is characterized by accumulation of aseptic fluid in the presence of a persistent corpus luteum. It forms a major cause of subfertility in goats. Active immunization against prostaglandin F2 alpha (PGF2 alpha), the luteolytic hormone in this species, was used to explore the possibility for artificial induction of this pseudopregnant condition. During the breeding season, 11 goats (group I) were immunized with 5 mg PGF2 alpha-ovalbumin conjugate in Freund's complete adjuvant, 4 goats were control-immunized (group CI) and 5 goats remained untreated (group C). Jugular blood samples were taken twice a week (monday and thursday) for measurements of plasma progesterone and binding of 3HPGF2 alpha. In conjunction with blood sampling, transcutaneous ultrasonographic examination of the uterus took place to detect the presence of fluid in the uterus. Before and immediately after immunization, the mean (+/-SD) duration of luteal phases (progesterone concentrations > or = 1 ng ml-1) was 16.7 +/- 1.6 (n = 39), 17.8 +/- 1.3 (n = 23) and 16.9 +/- 1.1 (n = 18) days in animals of group I, CI and C respectively. Ten goats of group I developed an antibody titre. Persistence of luteal function (mean duration +/-SD: 150.3 +/- 23.5 days) occurred in 6 of these animals and in 1 goat of group CI. Accumulation of fluid in the uterus in group I was first observed between day 31 and 38 of the prolonged luteal phase. Discharge of uterine fluid occurred as soon as the plasma progesterone concentration reached a level lower than 0.5 ng ml-1. It is concluded that immunization against PGF2 alpha is an effective method to induce pseudopregnancy in goats, providing a model for studies on luteal maintenance and uterine function in the absence of a conceptus.

Animals↗

Early diagnosis of enteroviral meningitis by a solid-phase reverse immunosorbent test and virus isolation.

45 cases of aseptic meningitis/meningoencephalitis were studied with regard to enteroviral etiology by virus isolation and solid-phase reverse immunosorbent test (SPRIST), a cross-reacting test for enterovirus IgM. An etiological diagnosis was reached in 37/45 (82%) patients. Etiological diagnoses other than enteroviruses were found in 8 patients: Borrelia burgdorferi in 4, varicella-zoster virus in 2, herpes simplex virus in 1 and mumps virus in 1 patient. Enteroviruses (echovirus 6, 21 and 30) were isolated from cerebrospinal fluid (CSF) in 26/37 (70%) and from stool samples of 20/21 (95%) of patients with no other etiology. Altogether enteroviruses were isolated from CSF and/or faecal samples in 29 patients. Echovirus 30 dominated as etiologic agent. In 34/40 (85%) of the samples with an enterovirus, a cytopathogenic effect was observed in cell culture within 4 days. In patients with an enterovirus isolate a SPRIST IgM response to echovirus 3, 5, 7 and/or coxsackievirus B3 was detected in 6/13 (46%) sera sampled 3-4 days after the onset of meningeal symptoms and in altogether 17/25 patients (68%). In 4 out of these virus isolation positive and SPRIST negative patients a single serum for SPRIST was available less than 4 days after onset of meningeal symptoms. Antigen from echovirus 5 gave the highest diagnostic yield. The SPRIST IgM test was positive in 2 cases where virus isolation, complement fixation and neutralization tests were negative. Epidemiological data however supported an enteroviral diagnosis in both of them. In conclusion, both SPRIST and virus isolation seem to be valuable for the early diagnosis of enteroviral meningitis.

Adolescent↗

Local hemorheological disorders during chronic inflammation.

Hemorheological parameters were investigated before, during, and after the standard burn of the distal part of the rabbits' ear shell (54 degrees C for 3 minutes). The erythrocyte aggregation was investigated with the "Georgian technique" and the local hematocrit was determined by centrifugation of blood samples from the inflammatory focus. In addition, we determined the size of the inflammatory edema in the ear shell. We found that the erythrocyte aggregability rised by 3.7 times in the venous blood flowing out from the inflammation focus while the hematocrit increased 1.6 times as compared to the microcirculation in the contralateral (control) ear shall. The thickness of this latter related to edema development increased three times. All the mentioned changes disappeared within five days after start of the experiments. We concluded that in the aseptic inflammatory foci the erythrocyte aggregability increases considerably in the microcirculation producing capillary stases and enhancing the local hematocrit. The hemorheological disorders were not spread to other parts of the circulatory bed.

Animals↗

[Bacteremia in intra-arterial angiography, percutaneous transluminal angioplasty and percutaneous transhepatic cholangio-drainage].

PURPOSE: Prospective evaluation of the rate of bacteremia attributed to invasive radiological techniques. METHODS: Aerobic and anerobic blood cultures were obtained in 100 patients (62 men, 38 women; mean age 65 +/- 14 years) undergoing intra-arterial angiography (N = 50), PTA (N = 30) or percutaneous transhepatic biliary drainage (PTCD; N = 20). Samples were taken before the treatment (T0), immediately after puncture of the vessel or bile duct (T2), and 30 min after the termination of the procedure (T3). RESULTS: The overall rate of bacteremia was 18%. During diagnostic angiography a 16% rate of temporary bacteremia (no positive T3 samples) was observed. During PTA the rate was 27% (no clinically significant infectious disease) and during PTCD the rate was 10% (5% cholangitis with septicemia). We isolated staphylococci (S. epidermidis: N = 7, S. species: N = 3, S. aureus: N = 1), streptococci (N = 2), Propionibacterium acnes (N = 5), E. coli (N = 1), Enterococcus faecium (N = 1), Enterobacter species (N = 1), and Clostridium perfringens (N = 1). Apart from the one patient with cholangitis no clinical infectious complication occurred. CONCLUSION: Temporary bacteremia is rather frequent during invasive radiological procedures. Strictly aseptic conditions and antibiotic prophylaxis, specially in case of implantation of a permanent foreign body, is warranted.

Aged↗

Infusion therapy team and dressing changes of central venous catheters.

OBJECTIVE: To determine whether central venous catheter (CVC) dressing changes could be performed by ward nurses rather than by the infusion therapy team (ITT) nurses without increasing the risk of catheter-related infection. DESIGN: Retrospective cohort study using prospectively collected data. The study extended from January 1995 to June 1996. SETTING: The University of Texas M.D. Anderson Cancer Center, a referral cancer center. PATIENTS: The study group was a random sample of 483 patients who received CVC dressing changes by ward nurses during the study period. A random sample of 483 patients who received CVC dressing changes by the ITT constituted the control group. RESULTS: The risks of catheter-related septicemia were 1.7% among cases and 1.4% among controls (risk ratio, 1.14; 95% confidence interval [CI95], 0.26-6.42; P=.70). There also were no significant differences between the two groups in the risks of catheter-related site infection (risk ratio, 0.50; CI95, 0.02-4.12; P=.25) or any catheter-related infection (risk ratio=1.00; CI95, 0.27-3.64; P=.59). CONCLUSIONS: Provided that aseptic techniques (including maximal barrier precautions during insertion) are maintained, the responsibility of CVC dressing changes could be delegated to the ward nurses without increasing the low risk of CVC-related infection, resulting in an estimated cost saving in excess of $90,000 per year.

Adult↗

[Determination of the elastic properties of the compact bone in the femur of dogs].

Total hip endoprotheses are a good possibility for treatment degenerative wear and pathologic damage of the hip joint in man as well as in dogs. However, aseptic loosening of the protheses, especially in the area of the shaft, is still a problem in conventional total hip endoprotheses. The purpose of the present study was to use the finite-element-analysis (FEA), to enhance endoprothetic design and to prevent loosening of protheses. In order to simulate the femur of the dog for numerical analysis, a material law for the compacta in the femur of the dog was developed. The elastic properties of the compacta were experimentally determined by using compression tests of bone samples of euthanised dogs. The results show constant denseness and a constant axial elastic modul in the compacta.

Animals↗

Rapid diagnosis of gram-negative bacterial meningitis by the Limulus endotoxin assay.

The Limulus amoebocyte lysate endotoxin assay was evaluated as a method for rapid diagnosis of acute bacterial meningitis in a series of 305 patients. The results of Limulus assays on cerebrospinal fluid (CSF) samples from these patients were compared with the results for each patient of routine bacterial cultures and Gram stains. Positive Limulus tests were obtained on initial CSF specimens from 84% of patients with culture-proven bacterial meningitis, including all patients with meningitis due to gram-negative organisms. Initial Gram-stained smears revealed the presence of organisms in 68% of the patients. One patient with pneumococcal meningitis had a weakly positive Limulus assay, whereas patients with meningitis due to other gram-positive organisms, those with aseptic meningitis, or patients without meningitis had negative CSF Limulus tests. The Limulus assay also demonstrated the persistence of endotoxin in the CSF of certain patients during antibiotic therapy, especially patients with Haemophilus influenzae meningitis. The Limulus test proved to be a rapid, reliable indicator of the presence of gram-negative organisms in the CSF of patients suspected of acute bacterial meningitis.

Adult↗

Tumor necrosis factor-alpha (TNF-alpha) in loosening of total hip replacement (THR).

OBJECTIVE: The initially well-fixed implants of total hip replacement (THR) are in the long-term subject to aseptic loosening. Many cytokines can contribute to osteolysis due to osteoclast recruitment and/or activation. However, in this respect tumor necrosis factor-alpha (TNF-alpha) plays a pivotal role, because it upregulates interleukin-1 and 6 and granulocyte-macrophage colony stimulating factor. The aim of this study was to assess the eventual presence, cellular localization and extent of expression of TNF-alpha in the synovial-like membrane at the implant or at the cement to bone interface compared to control synovial membrane. METHODS: Twenty samples from the synovial-like membrane of the periprosthetic tissues were compared to control samples. TNF-alpha containing cells were visualized using an avidin-biotin-peroxidase complex (ABC) method and analyzed by light microscopy, double labelling and image analysis. RESULTS: TNF-alpha was found in the periprosthetic tissues in fibroblasts and vascular endothelial cells, but mainly in the macrophages was it found to coincide with areas containing implant-derived debris. TNF-alpha containing cells were more numerous in the synovial-like membrane in the interface tissue from the proximal stem area (2816 +/- 318 cells) than in the control synovial membrane (565 +/- 93 cells, p < 0.01). Interestingly, similarly high TNF-alpha expression (3452 +/- 582 cells) was also seen in the synovial-like membrane of the pseudocapsule. CONCLUSION: These findings suggest that the foreign body-type host reaction caused by THR is characterized by the high expression of TNF-alpha. Because such expression occurred in the interface tissue between the implant and surrounding bone, TNF-alpha, due to its pivotal direct and indirect role in the activation and recruitment of osteoclasts, may contribute to periprosthetic osteolysis and to the loosening of THR.

Adult↗

Carriage of oral flora on irreversible hydrocolloid and elastomeric impression materials.

This study assessed the carriage and persistence of oral flora on irreversible hydrocolloid and elastomeric impression materials. In the first part of the study, a single isolate each of Streptococcus mutans, Escherichia coli, Staphylococcus aureus, and Candida albicans were inoculated onto each of two elastomeric and two irreversible hydrocolloid impression surfaces artificially created in vitro. Then the inoculum was discarded, samples of impression material (13 mm in diameter) were removed over a 5-hour period, and the remainder of viable organisms were estimated. In the second part of the study, impressions were made from 21 dentate and eight edentulous patients, and bacterial counts were estimated as described. The results showed twofold to fivefold retention of bacteria on the irreversible hydrocolloids compared with the elastomeric impression surfaces. With all impression materials, the number of viable bacteria retained within 5 hours decreased 65% to 98% except for the "disinfectant" irreversible hydrocolloid Blueprint Asept, which totally destroyed the organisms in less than 3 minutes. It is concluded that the total bacterial "load" on impression surfaces is relatively low and decreases rapidly after impression making. However, disinfection of impression surfaces should be mandatory to prevent cross-infection.

Adult↗

Persistence of Mycoplasma bovis infection in the mammary glands of lactating cows inoculated experimentally.

To study the course of clinical mycoplasma mastitis and investigate its potential for persistence, 10(8) colony-forming units (cfu) of an Irish isolate of Mycoplasma bovis was inoculated aseptically into the right fore teat canal of three lactating cows. M bovis rapidly colonised the infected quarters and grew exponentially to more than 10(10) cfu/ml within the first three days, and spread to other quarters of each of the three cows within five to 10 days. After periods of between 24 and 72 hours the infected quarters became distended and sensitive to touch, and their secretions changed from containing visible particles, to a seropurulent exudate, to an aqueous suspension of fine particles which formed a sediment after a sample was collected. M bovis-specific antibody levels increased to varying degrees in all three cows. Subsequently, the concentrations of mycoplasma decreased to less than 10(7) cfu/ml in two of the cows, but remained at more than 10(8) cfu/ml to the end of the lactation of the other cow. Apparently normal milk was secreted by one of the cows within a month of the challenge, and by the other two cows at the start of their next lactation. However, in two of the cows subclinical M bovis infection persisted through the dry periods and into their next lactations.

Animals↗

Osteotomy with 80-micros CO2 laser pulses--histological results.

Haemostatic and aseptic effects and intricate cut geometry are beneficial aspects of non-contact laser osteotomy. Collateral thermal damage, however, has severely limited the use of conventional lasers. The purpose of this study was to test the side effects on bone after cutting it with short CO2 laser pulses and simultaneous application of a fine air-water spray. The 10.6 microm CO2 laser emitted 80 micros pulses of 46 mJ energy, f=100 Hz, focused to a spot diameter of 130 ìm. Scan rate amounted to 40 mm/s. To approximate live conditions 10 samples of cortical bone and 10 rib segments were prepared immediately after sacrificing of pigs. A reference cut with a bandsaw and three laser cuts with an increasing number of beam passes (4, 16, 64) were performed on each sample. Half of the samples were decalcified in EDTA. The others were embedded in plastic to cut non-decalcified sections. The laser incisions were not accompanied by carbonisation. The incisions with slightly convergent walls were 150 ìm wide. The depths of the cavities increased with the number of the beam passes from approximately 0.5 mm (4 passes) to 3 mm (64 passes). At the border of the incisions two narrow zones of damage were noted: an amorphous intensively stained zone of 1-3 microm width and a wider, also sharply demarcated but faintly stained zone of 7-10 microm. A broader zone of about 50 microm was characterised by empty lacunae and osteocyte damage. These effects were not predictable; intact osteocytes were also observed near to the cut surface. Polarised light microscopy showed no alterations in the inorganic structure of the bone at the cut borders. The histological results indicated only minimal damage to bone ablated at the specified parameters. The described laser procedure might have advantages over mechanical instruments.

Animals↗