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PubMed2001

Prevalence of campylobacter spp. from skin, crop, and intestine of commercial broiler chicken carcasses at processing.

This study describes the prevalence of positive Campylobacter cultures from the skin, crop, and intestine of postscald broiler chicken carcasses at processing. Six to 12 carcasses from 22 flocks were sampled. Skin was cultured by direct plating of a cotton swab, whereas crop and intestine were cultured from tissue that was aseptically harvested and stomached in PBS before plating. Cultures were not enriched prior to plating. The methods used in this report are compared to those used by others. In this study, skin samples were 78% positive; crops were 48% positive, and intestines were 94% positive (n = 202). Based on our results, if the intestine was positive for Campylobacter, the odds of finding a positive crop culture was 8.6 times greater, and the odds of finding a positive skin culture was 35 times greater than if the intestinal culture was negative for Campylobacter. These data suggest that the intestine was the most likely organ of those tested to be positive in postscald broiler carcasses from positive flocks. Further, if only one organ can be sampled, intestinal samples are most likely to reflect the prevalence of Campylobacter in a flock.

Animals↗
PubMed1985

A method for the collection of defined areas from the embryonic rat brain for cell and tissue culture.

Tissue for the culture of cerebral neurons has frequently been taken from brains of the embryonic rats. In many cases it was impossible to obtain regularly and reproducibly small, defined pieces of tissue, e.g. diencephalic nuclei, of the extremely soft embryonic cerebral tissue. On the other hand, as a basis for tissue cultures, well defined samples are more and more considered essential. Therefore a method for collecting samples of defined small regions from the di- and mesencephalic rat brain, 17 days of gestation, was developed. It is applicable to cell and tissue culture. Embryonic brains are prepared aseptically and embedded in congealing Agarose. Stabilized in this gel they are cut into 225 micron slices using a tissue chopper. Tissue samples desired for culture are then punched out from the respective brain slice, which previously had been compared with corresponding reference micrographs. The correct localization of the tissue punch within the fresh brain slice is controlled histologically. The embedding procedure for performing the histology of the brain slices of 225 micron width is described. For application of the introduced method, coordinates for slicing and histological reference micrographs are given for di- and mesencephalic areas.

Animals↗
PubMed1998

Rapid detection of enteroviral RNA in cerebrospinal fluid (CSF) from patients with aseptic meningitis by reverse transcription-nested polymerase chain reaction.

The aim of this study was to compare conventional enterovirus isolation with rapid detection of enteroviral RNA by a reverse transcription-nested polymerase chain reaction (RT-nPCR) method amplifying the 5' nontranslated region of the enteroviral genome in specimens from patients with aseptic meningitis. Reference enterovirus strains and clinical enterovirus isolates were analyzed to evaluate assay sensitivity and specificity. All known enteroviral serotypes tested, but one (echovirus type 22), were detected by RT-nPCR. A series of unrelated viral isolates as well as CSF samples from patients with meningitis/encephalitis or neurological syndromes unrelated to enterovirus infection were included as controls. A total of 47 specimens (31 CSF, 12 rectal swabs, 4 throat swabs) from 30 patients with aseptic meningitis were available for the study. Of the 31 CSF samples tested from 30 patients, 17 from 17 patients (54.8%) were positive by RT-nPCR, while only 10 from 10 patients (32.2%) were positive by culture. Thus, RT-nPCR allowed diagnosis of enterovirus meningitis in 7 additional patients compared to cell culture. The cytopathic effect was observed 5-15 days after inoculation of CSF specimens onto cell cultures, while direct detection of viral RNA in CSF samples by RT-nPCR permitted diagnosis of enteroviral meningitis within 1-2 days. On the whole, viral isolation was positive in 12/47 (25.5%) specimens, whereas viral RNA was detected by RT-nPCR in 11 additional samples (23/47, 48.9%). Specimens of the control group were consistently negative by both viral isolation and RT-nPCR. Restriction endonuclease analysis of PCR products (RFLP) was applied to differentiate poliovirus (PV) from non-polio enteroviruses (NPEV). All enterovirus strains detected in clinical samples (n = 23) were identified as NPEV by RFLP. Clinical isolates were typed by neutralization as echovirus type 30 (n = 6), while 6 were not typed. In conclusion, detection of enteroviral RNA in CSF by RT-nPCR allows: i) rapid diagnosis of enteroviral meningitis; ii) increased sensitivity with respect to virus isolation; iii) differentiation between PV and NPEV infections of the central nervous system.

Adolescent↗
PubMed2003

Mollaret's meningitis: cytopathologic analysis of fourteen cases.

Mollaret's meningitis (MM) is a rare disease of benign nature characterized by recurrent episodes of aseptic meningitis. Cerebrospinal fluid (CSF) examination remains the sole diagnostic modality. Eighteen CSF samples from 14 patients were studied along with the clinical data. Specimens were prepared by cytocentrifugation and Millipore filtration and were stained with Diff-Quik and Papanicolaou stains. Eight patients were men and six were women, with an age range of 17-74 yr (mean age 37 yr). Most common clinical presentation was recurrent episodes of headaches and photophobia followed by a sustained mild fever lasting 5-7 days. The CSF showed markedly increased cellularity with pleocytosis. The differential count showed predominant monocytosis ranging from 84% to 100% (mean 96). In our series, two patients had herpes simplex virus type 2 (HSV-2) DNA detected by polymerase chain reaction (PCR) in the CSF. The monocytes were seen predominantly singly, but three cases showed a strong tendency to aggregate in small groups. Phenotypically, these cells had bean-shaped bilobed nuclei as well as multiple deep nuclear clefts depicting the so-called "footprint" appearance. In four cases, multiple blunt-tipped cytoplasmic pseudopods were noted. Degenerated monocytes with the appearance of the so-called "ghost cells" were noted in one-half of the cases. Background cells were mostly small mature lymphocytes; however, one-half of cases showed a significant amount of plasma cells and/or polymorphonuclear leukocytes (PMNs). Lysed blood with hemosiderin-laden macrophages and numerous leptomeningeal cells were seen in two cases. CSF examination of MM presents a spectrum of cytomorphologic features. When interpreted in light of the appropriate clinical setting. the latter, although nonspecific, provides an accurate diagnosis. The differential diagnosis includes various degenerative, inflammatory/infectious, and lymphoproliferative disorders of the central nervous system.

Adolescent↗
PubMed2001

Correlation of synovial fluid cytokine levels with histological and clinical parameters of primary and revision total hip and total knee replacements.

We retrieved synovial tissue and fluid samples from patients undergoing primary total hip replacement (THR) (n 15), revision of aseptically loose THR (n 12), primary total knee replacement (TKR) (n 13) and revision of aseptically loose TKR (n 6). Several histological parameters were assessed on a relative scale of 14. Primary TJRs were clinically evaluated for degree of osteoarthrosis. Revision TJRs were assessed for migration of the implant, gross loosening and the degree of radiolucency. Cytokine levels in synovial fluid were determined with ELISA. All cytokines were significantly higher in revision TJRs than in primary replacements, as were the degree of macrophage and giant cell infiltration. We found no relationship between any clinical variable and the levels of any cytokine, but migration of the implant was related to the presence of PE debris. A significant correlation was seen between the presence of macrophages and the levels of IL-1beta, IL-8 and IL-10, but not IL-6. No differences were noted between hips and knees for any of the variables, except in the levels of IL-6, where higher levels were found in THRs. These results suggest a unique role for IL-6 that requires further investigation.

Adult↗
PubMed2005

Use of autologous serum in corneal epithelial defects post-lamellar surgery.

PURPOSE: To evaluate the efficacy of an autologous serum treatment of post-LASIK (laser in situ keratomileusis) corneal epithelial defects in a rabbit model. METHODS: Five milliliters blood samples from 10 New Zealand rabbits were obtained by venepuncture. The serum was aseptically separated and diluted with saline solution to 20%. The final preparation was placed into 3-mL bottles with ultraviolet protection and maintained at 4 degrees C. Corneas were de-epithelialized using a 7-mm optical zone marker. A 160-microm thick flap was created in both eyes of all rabbits using an automatic corneal shaper microkeratome. Right eyes were treated with serum drops 6 times per day. Left eyes were treated with preservative-free artificial tears. Vital staining of the ocular surface and the area of corneal epithelial defect was measured daily for 1 week. Rabbits were humanely euthanized at postoperative day 7, and corneas were fixed and sectioned. Hematoxylin and eosin staining and immunohistochemical analysis were performed. RESULTS: Corneas treated with autologous serum had a statistically significant increase in the epithelial healing rate compared with those treated with artificial tears. Serum-treated corneas showed significantly less terminal transferase-mediated dUTP nick-end labeling (TUNEL) staining in the interface, minimal inflammatory cell infiltration, and less induced synthesis of stromal chondroitin sulfate than did corneas treated with preservative-free artificial tears. CONCLUSIONS: Treatment with autologous serum could be an efficient way to provide essential components to the ocular surface in the treatment of post-LASIK epithelial defects. Autologous serum induces faster epithelial healing than do artificial tears, leading to (1) a decrease in keratocyte apoptosis and migration of fibroblasts and myofibroblasts in the wound site, (2) a decrease in the migration of inflammatory cells, and (3) a consequent inhibition of cytokine release. This treatment could improve long-term refractive results post-LASIK.

Animals↗
PubMed2001

West Nile encephalitis in Russia 1999-2001: were we ready? Are we ready?

In 1963-1993, several strains of West Nile virus (WNV) were isolated from ticks, birds, and mosquitoes in the southern area of European Russia and western Siberia. In the same regions, anti-WNV antibody was found in 0.4-8% of healthy adult donors. Sporadic human clinical cases were observed in the delta of the Volga River. In spite of this, WNV infection was not considered by the health authorities as a potentially emerging infection, and the large WNV outbreak in southern Russia, started in late July 1999, was not recognized in a timely fashion. First evidence suggesting a WNV etiology of the outbreak was obtained by IgM ELISA on September 9. Two weeks later, the specific WNV RT-PCR was developed and WNV disease was confirmed in all 14 nonsurvivors from whom brain tissue samples were available. Retrospective studies of serum samples by IgM ELISA indicated WNV etiology in 326 of 463 survivors with aseptic meningitis or encephalitis. Moreover, 35 of 56 patients who contracted aseptic meningitis in 1998 had a high titer of WNV IgG antibody, so the WNV infection seems to have been introduced into the Volgograd region before 1999. A complete sequence (AF317203) of WN viral RNA, isolated from the brain of one Volgograd fatality, and partial sequences of an envelope E gene from other nonsurvivors showed that the Volgograd isolate had the greatest homology (99.6%) with WN-Romania-1996 mosquito strain RO97-50.

Age Distribution↗
PubMed1995

Interleukin-6 in cerebrospinal fluid of patients with central nervous system infections.

Interleukin(IL)-6 levels were measured in cerebrospinal fluid (CSF) and serum samples from pediatric patients with central nervous system (CNS) infections by means of an enzyme-linked immunosorbent assay. Mean IL-6 concentrations in CSF samples from patients with bacterial meningitis (49,017 +/- 44,730 pg/ml) were significantly higher than those in patients with aseptic meningitis (1076 +/- 1572 pg/ml) or encephalitis (409 +/- 835 pg/ml). In aseptic meningitis and encephalitis, IL-6 levels in serum were within the lower ranges (< 100 pg/ml), in contrast with the highly elevated levels found in bacterial meningitis (14,332 +/- 18,385 pg/ml). In 5 of the 15 patients with encephalitis, elevated levels of IL-6 were observed in the initial CSF samples despite normal findings of routine CSF examinations. Also, sequential CSF samples revealed that there was an increase in the CSF cell count in two of the five patients. These results validated the potential of measuring IL-6 in CSF samples for the purpose of providing additional information on routine laboratory test results.

Blood-Brain Barrier↗
PubMed2005

Rapid and sensitive detection of mumps virus RNA directly from clinical samples by real-time PCR.

A rapid, sensitive, and specific assay to detect mumps virus RNA directly from clinical specimens using a real-time PCR assay was developed. The assay was capable of detecting five copies of standard plasmid containing cDNA from the mumps virus F gene. No cross-reactions were observed with other members of Paramyxoviridae, or with viruses or bacteria known to be meningitis pathogens. Seventy-three clinical samples consisting of throat swabs collected from patients with parotitis, and cerebrospinal fluid (CSF) collected from patients with aseptic meningitis, were examined with a real-time PCR assay developed by the authors, reverse-transcription nested-PCR (RT-n-PCR), and virus isolation using cell culture. Like the RT-n-PCR assay, the real-time PCR assay could detect mumps virus RNA in approximately 70% of both throat swabs and CSF samples, while, by tissue culture, mumps virus was isolated from only approximately 20% of CSF and 50% of throat swab samples. In addition, the real-time PCR assay could be developed easily into a quantitative assay for clinical specimens containing more than 1,800 copies of mumps virus RNA/ml by using serial dilutions of the standard plasmid. The results suggest that the real-time PCR assay is useful for identification of mumps virus infections, not only in typical cases, but also in suspected cases, which show only symptoms of meningitis or encephalitis.

Humans↗
PubMedDate not supplied

Cerebrospinal fluid antiganglioside antibodies in patients with AIDS.

In this study the presence of brain antiganglioside antibodies in the cerebrospinal fluid (CSF) of patients with HIV infection was analysed. CSF samples were collected from 45 patients with AIDS and from 45 anti-HIV negative subjects, 15 of whom presented aseptic meningitis. Nineteen AIDS patients had clinically well-documented encephalopathy. Thirteen of these patients had white matter lesions shown by magnetic resonance imaging (MRI). Both IgG and IgM antiganglioside antibodies were detected by immunostaining on thin layer chromatography plates in three CSF samples from AIDS patients with progressive encephalopathy with signs of a diffuse demyelination, as revealed by MRI. Two of these CSF samples reacted specifically with GM3, GM1 and GD1a and one with GD1a. In none of the HIV infected patients without demyelinating encephalopathy, but with opportunistic infections or cerebral lymphoma, nor in the anti-HIV negative control subjects were antiganglioside antibodies detected. No association with JCV DNA, CMV DNA, EBV DNA, detected by nested PCR, nor HIV antigen p24 was found. These findings show the presence of brain antiganglioside antibodies in the CSF of AIDS patients for the first time. However, the findings do not suggest relating the presence of these antibodies to HIV encephalopathy or particular viral agents, but indicate that the antibodies are detectable in subjects with progressive encephalopathy with a diffuse demyelination.

Acquired Immunodeficiency Syndrome↗
PubMed1983

A nursing practice problem: failure to observe aseptic technique.

A study was conducted to assess nurses' adherence to a recently revised policy and procedure manual. Medical-surgical nurses (n = 217) were surveyed to assess their perceived ability to perform selected clinical procedures. A sample (n = 34) of respondents was then observed performing the procedures. Analysis of the 37 errors in performance revealed an unanticipated high number of errors (n = 33) in aseptic technique. Type of errors and projected reasons for their occurrence are given.

Antisepsis↗
PubMed2003

[FDG PET as an early nuclear medical diagnostic tool for aseptic wear-induced loosening of hip joint endoprostheses--a report of two cases].

To date, 2-[F-18]fluoro-2-deoxy-D-glucose PET (FDG PET) is used as a tool in oncology as well as myocardiological and cerebral functional diagnostics in the clinical routine. False positive results of tumor search imply new possibilities for use in diagnostics of inflammation. The two case reports presented here on aseptic loosening of endoprostheses caused by the rub of polyethylene with histological and immunohistological refurbishing call attention to the possible diagnostic valency of FDG PET as an early warning system regarding aseptic artificial limb loosening induced by the rub of polyethylene. By quantification of glucose metabolism with the "standard uptake value" (SUV) as well as specific storage samples from around the artificial limb, it is the aim of our study group in further examinations to develop an algorithm which permits to distinguish between septic and aseptic loosening.

Acetabulum↗
PubMed2006

A preliminary survey of the arbuscular mycorrhizal status of grassland plants in southern Tibet.

We report for the first time the arbuscular mycorrhizal (AM) status of native plant species and AM fungal diversity in the grasslands of southern Tibet. A total of 51 soil samples were collected from the rhizospheres of the dominant plant species, and AM fungal structures were observed in 18 (82%) of 22 plant species examined. Vesicles and aseptate hyphae were the structures most frequently observed in the plant roots. After trap culture for 5 months, 25 AM fungal taxa were identified in the soil samples collected, of which nine belonged to Glomus, ten to Acaulospora, one to Entrophospora and five to Scutellospora. The frequency of occurrence of different genera and species varied greatly. Glomus was the dominant genus, and the most frequent and abundant species was Glomus mosseae. Over the whole sampling area, spore density in the rhizosphere soil of different host plant species ranged from 2 to 66 per 20 g air-dried soil. Overall AM fungal species richness was 2.10 and species diversity was 2.35. AM fungal diversity was also compared among the four different land use types (farmland and normal, disturbed and highly disturbed montane scrub grassland). Spore densities in the farmland and normal grassland were much higher than in the grasslands that had been degraded to varying degrees. The species richness in normal grassland was the highest of the four land use types examined. Species diversity varied from 1.99 to 0.94 and was highest in normal grassland, intermediate in degraded grassland and farmland, and lowest in the highly disturbed grassland.

Biodiversity↗
PubMedDate not supplied

Short-term aseptic loosening of the femoral component in canine total hip replacement: effects of cementing technique on cement mantle grade.

OBJECTIVES: To evaluate the effects of different cementing techniques on radiographic cement mantle grade and short-term aseptic loosening of the femoral component in canine total hip replacement (THR). STUDY DESIGN: Retrospective study. SAMPLE POPULATION: Radiographs of 284 dogs that had THR. METHODS: Immediate postoperative radiographs of 284 dogs that had cemented THR were reviewed by 4 surgeons and 1 radiologist and assigned a cement mantle grade using a 4-grade classification system. Dog age and weight at surgery, cementing technique (1st, 2nd, 3rd generation), complications (type and timing), and follow-up time were retrieved and analyzed. RESULTS: Mean cement mantle score for 2nd generation technique was significantly higher than 3rd generation technique and both were significantly higher than 1st generation technique (P<.001). Aseptic loosening was the 2nd most common complication observed with an incidence of 2.1%. Mean time from surgery to last follow-up examination was 122 days. No statistically significant difference in incidence of aseptic loosening was identified among different cement mantle grades or cementing techniques. CONCLUSIONS: Advanced cementing techniques resulted in better cement mantles based on grading of immediate postoperative radiographs, however grading did not predict short-term aseptic loosening. Cementing technique seemingly did not affect the incidence of short-term aseptic loosening of the femoral component for dogs in this study. CLINICAL RELEVANCE: Our study suggests that advances in cementing technique may result in improvements in the radiographic grade of cement mantles. With respect to aseptic loosening of the femoral component, our data only suggest that short-term (3 months-3 years) loosening cannot be predicted by immediate postoperative radiographic evaluation of cement mantle.

Animals↗
PubMed2000

Managing central venous catheters: a prospective randomised trial of two methods.

A randomised, prospective study was conducted to evaluate the impact on central venous catheter (CVC) infection when fluids and lines connected to a CVC were changed using a 'sterile' compared to an 'aseptic, non-touch' technique. The study sought to determine whether there were any differences in CVC tip colonisation (CTC) or CVC-related bacteraemia (CRB) as a result of the technique used for fluid and line changes. In the sterile technique (control) group, fluids and tubing were changed using full sterile technique. In the aseptic, non-touch (experimental) group, fluids and tubing attached to the CVC were changed using only a small sterile drape and a 2-minute clinical hand wash. When the CVC was removed, the tip was sampled and cultured using the semi-quantitative method. Blood cultures were also collected. In all, 111 samples from 79 patients were included in the trial: 61 in the sterile technique group and 50 in the non-touch, aseptic technique group. Results showed a CTC rate of 31 per cent in the control group and 14 per cent in the experimental group, while the CRB rate was 8.2 per cent and 6 per cent respectively. The most common organisms cultured were Staphylococcus aureus and S. epidermis respectively. This study indicates that it is safe to change fluids and lines attached to CVCs using the aseptic, non-touch technique, which has resulted in significant financial savings through less use of equipment and less nursing time required to perform fluid and line changes.

Bacteremia↗
PubMed2000

Intestinal bacteria affect growth of Bacillus thuringiensis in larvae of the oriental tea tortrix, Homona magnanima diakonoff (Lepidoptera: tortricidae).

Spores and parasporal crystals of a Bacillus thuringiensis serovar aizawai were fed to fifth instar larvae of the oriental tea tortrix, Homona magnanima, that had been reared aseptically or that had been reared normally. Viable cell numbers of B. thuringiensis and other bacteria in H. magnanima larvae were estimated by homogenization of samples and dilution plating on peptone-polymyxin agar medium for B. thuringiensis cells and on nutrient agar medium for the other bacterial cells. B. thuringiensis did not grow in the larval cadavers of normally reared H. magnanima while bacteria other than B. thuringiensis grew rapidly. In contrast, B. thuringiensis within the larval cadavers of aseptically reared H. magnanima grew and increased 20 times. The bacteria other than B. thuringiensis from the sample homogenates of normally reared larvae that were fed on B. thuringiensis-treated diets had the same characteristics as the bacteria isolated from the guts of healthy H. magnanima larvae, which were putatively identified as Streptococcus spp. and Staphylococcus spp., typical intestinal bacteria of insects. The results strongly suggest that intestinal bacteria influence the growth of B. thuringiensis in the larvae.

Animals↗
PubMedDate not supplied

Pharmacokinetics of caffeine in breast milk and plasma after single oral administration of caffeine to lactating mothers.

Six healthy lactating women ranging in age from 28 to 35 years were selected to participate in the study. The women had been nursing their infants from 3 1/2 weeks up to 17 weeks. On the morning of the study each participant received 100 mg dose of caffeine. The oral dose of caffeine was taken with a glass of water on an empty stomach. Blood samples were collected from zero time to 24 h after caffeine administration. Each subject aseptically collected breast milk from the right and left breasts by manual expression or using a mechanical breast pump, from zero time to 24 h after caffeine administration. Caffeine in plasma and breast milk was determined by gas liquid chromatography using nitrogen phosphorous detection. Caffeine was rapidly absorbed producing plasma time to peaks ranging from 0.50 to 1.00 h, with values for peak concentrations between 3.60 and 6.15 micrograms ml-1 in plasma. In breast milk, time to peak ranged from 0.75 to 2.00 h and concentration values between 1.98 and 4.30 micrograms ml-1. No significant differences were found between attainment of the time of the peak in plasma and breast milk, p greater than 0.05. In addition, peak concentration for the right and left breast were not statistically different, p greater than 0.05. However, significant differences, p less than 0.05, between plasma and both breasts in regards to the peak concentrations were found. The overall breast milk/plasma ratio obtained from the respective area under the curves were 0.815 +/- 0.0521 and 0.809 +/- 0.202 for the right and left breast, respectively, and no statistical differences were observed between the right and left breasts, p greater than 0.05. In summary, caffeine is rapidly transferred to breast milk. Multiple dose studies are needed to provide more information about caffeine concentration in breast milk of women who receive multiple doses of caffeine or who chronically consume caffeine.

Administration, Oral↗
PubMedDate not supplied

Diffusion-weighted images in children with meningoencephalitis.

PURPOSE: The objective of the study was to evaluate the maps of apparent diffusion coefficients (ADCs) and diffusion-weighted (DW) images in demonstrating meningoencephalitic lesions in children. MATERIALS AND METHODS: Between May 1998 and May 2000, 18 infants and children (4.5-190 months old) suffering from meningoencephalitis were included in the study. The diagnoses were bacterial meningoencephalitis in 8 and aseptic or viral in 10 patients. All 18 patients had brain MRI examinations. In the axial plane, three pulse sequences were performed on all patients: (1) FSE T2W images; (2) fast FLAIR images; (3) single-shot echoplanar DW images were acquired. Another 18 patients from the control group also received DW image examination. ADCs were computed for all regions on each DW image. RESULTS: The absolute values of CNRs of lesions on T2W (7.27+/-5.51), FLAIR (5.56+/-5.03) and DW (13.36+/-16.64) images were significantly greater than those on ADC maps (0.42+/-0.30) in the study group of patients (P<.01). In addition, absolute CNRs on DW images were significantly greater than on T2W and FLAIR images (P<.01). However, lesions on ADC maps in the study group have significantly greater CNRs than in the control group (0.13+/-0.12) (P<.01). CNRs on initial DW images from patients with atrophy or swelling of meningoencephalitic lesions were significantly different from the CNRs of those patients without significant changes in meningoencephalitic lesions (P=.02<.05). CONCLUSION: The DW image is a sensitive tool for detecting meningoencephalitic lesions and is better than FSE T2W and fast FLAIR images in CNRs. Diffusion MR techniques provides new ways to possibly predict the outcome of intracranial infectious diseases in children.

Brain Mapping↗