Facilitation of Corticospinal Excitability According to Motor Imagery and Mirror Therapy in Healthy Subjects and Stroke Patients

OBJECTIVE: To delineate the changes in corticospinal excitability when individuals are asked to exercise their hand using observation, motor imagery, voluntary exercise, and exercise with a mirror. METHOD: The participants consisted of 30 healthy subjects and 30 stroke patients. In healthy subjects, the amplitudes and latencies of motor evoked potential (MEP) were obtained using seven conditions: (A) rest; (B) imagery; (C) observation and imagery of the hand activity of other individuals; (D) observation and imagery of own ipsilateral hand activity; (E) observation and imagery of the hand activity of another individual with a mirror; (F) observation and imagery of own symmetric ipsilateral hand activity (thumb abduction) with a mirror; and (G) observation and imagery of own asymmetric ipsilateral hand activity (little finger abduction) with a mirror. In stroke patients, MEPs were obtained in the A, C, D, E, F conditions. RESULTS: In both groups, increment of the percentage MEP amplitude (at rest) and latency decrement of MEPs were significantly higher during the observation of the activity of the hand of another individual with a mirror and during symmetric ipsilateral hand activity on their own hand with a mirror than they were without a mirror. In healthy subjects, the increment of percentage MEP amplitude and latency decrement were significantly higher during the observation of the symmetric ipsilateral hand activity with a mirror compared to the observation of the activity of the asymmetric ipsilateral hand with a mirror of their own hand. CONCLUSION: In both groups, corticospinal excitability was facilitated by viewing the mirror image of the activity of the ipsilateral hand. These findings provide neurophysiological evidence supporting the application of various mirror imagery programs during stroke rehabilitation.

Escherichia coli pap Genes as well as Adenovirus Type 11 and Type 21, and BK Virus were Involved with Severe Urinary Tract Infection in Infants

In infants, urinary tract infections (UTIs) are quite common and primarily caused by bacterial pathogens. However, little research has been conducted regarding the relationship between uropathogenic bacteria, virulent genes, and uropathogenic viruses that might induce UTIs in infants. In this study, we evaluated infants with UTIs to determine the influence of bacterial virulent genes and type of viral infections on clinical aspects. First, we detected 44 cases of bacterial UTI from 600 suspected cases in infants and children. We detected E. coli urovirulence genes (kps, usp, pap, ireA, and cnf), two enteropathogenic E. coli genes (bfpA, and eae) and four S. aureus and S. epidermidis genes (mecA, pvl, bbp, and icaA) in urine samples from infant UTI cases. We also simultaneously detected hematuria-related adenovirus type 11, 21, and BK virus (BKV) in urine samples by PCR. As a result, E. coli was the most prevalent bacteria and in Dimercaptosuccinic acid (DMSA)-positive UTI cases, the uropathogenic E. coli virulence factor pap was significantly high. We found that BKV detection was significantly higher in DMSA-positive UTI infants (89%) compared with 50% of non-UTI (no bacteria detected) cases. These results are indicative of combined multiple bacterial and viral infections and show severe infant pyelonephritis.

Detection of Virulence Genes of Staphyloccus aureus and Staphylococcus epidermidis Isolated from Suprapubic Urine from Infants with Fever

While methicillin-resistant Staphylococcus aureus (MRSA) isolated from urinary tract infection (UTI) has recently increased in elderly adult urology patients, it has been only rarely reported in infants. Therefore, in this study to understand whether MRSA may be involved in UTI of infants who run fever without other apparent causes, we identified and counted S. aureus and S. epidermidis in suprapubic urine from 750 febrile infants via microbiological methods, and confirmed the counts via multiplex PCR. And we also detected four virulence genes, mecA, PVL, bbp and icaA genes for S. aureus and S. epidermidis via multiplex PCR in the same specimens. S. aureus (28 cases) counts were as follows: >10(4) CFU/ml (3/28), 10(2)~10(3) CFU/ml (1/28) and 10(4) CFU/ml (2/26), 10(2)~10(3) CFU/ml (4/26) and 10(2)~10(3) CFU/ml (20/26). S. aureus virulence genes were detected in 26 cases as mecA (16/26, 59.3%), PVL (17/26, 63.0%), bbp (7/26, 26.9%) and icaA (20/26, 76.9%). S. epidermidis virulence genes were detected in 22 cases as mecA (17/22, 81.0%), PVL (15/22, 71.4%), bbp (3/22, 13.6%) and icaA (13/22, 50.1%). Therefore, mecA, PVL and icaA genes of MRSA and MRSE were detected with high positivity in urines from infants with fever. The results demonstrate that community-acquired MRSA or MRSE may be responsible for UTI incidence in febrile infants.

Influence of Associated Medical Diseases and Complications on Functional Improvement after Rehabilitation in Patients with Stroke

OBJECTIVE: To investigate the influence of associated medical diseases and complications on functional improvement after in-patient through stroke rehabilitation. METHOD: We performed a retrospective analysis on medical records of 183 stroke patients who had admitted to the department of rehabilitation medicine. Functional Independence Measure (FIM), Modified Barthel Index (MBI) at admission and discharge were used to assess the functional status. We investigated medical diseases, such as hypertension, diabetes, myocardial infarct, atrial fibrillation osteoarthritis, rheumatoid arthritis, previous history of stroke and complications such as dementia, post-stroke depression, central post-stroke pain, complex regional pain syndrome, neglect and aphasia. RESULTS: Post-stroke patients with myocardial infarct, atrial fibrillation, osteoarthritis, dementia, aphasia and neglect significantly showed lower gain of FIM and MBI, lower FIM and MBI efficacy during inpatient rehabilitation compared to without those (p0.05). Total numbers of associated medical diseases and complications negatively affect on FIM and MBI efficacy (p<0.05). CONCLUSION: Therefore, it may be important to early detect and manage associated medical diseases and complications in post-stroke patients during rehabilitation, which improve the overall functional recovery of the patients.

Expressed Sequence Tags of Trichinella spiralis Muscle Stage Larvae

In order to obtain greater insight into the relevant genomic expression patterns of Trichinella spiralis, 992 expressed sequence tags (ESTs) were collected from a cDNA library of T. spiralis muscle stage larvae and assembled into 60 clusters and 385 singletons. Of them, 445 (44.7%) ESTs were annotated to their homologous genes, and small fractions were matched to known genes of nematodes. The annotated ESTs were classified into 25 eukaryotic orthologous groups (KOG). Cytochrome C oxidase (34 clones) was found to be most frequent species.

Differential Signaling via Tumor Necrosis Factor-Associated Factors (TRAFs) by CD27 and CD40 in Mouse B Cells

BACKGROUND: CD27 is recently known as a memory B cell marker and is mainly expressed in activated T cells, some B cell population and NK cells. CD27 is a member of tumor necrosis factor receptor family. Like CD40 molecule, CD27 has (P/S/T/A) X(Q/E)E motif for interacting with TNF receptor-associated factors (TRAFs), and TRAF2 and TRAF5 bindings to CD27 in 293T cells were reported. METHODS: To investigate the CD27 signaling effect in B cells, human CD40 extracellular domain containing mouse CD27 cytoplamic domain construct (hCD40-mCD27) was transfected into mouse B cell line CH12.LX and M12.4.1. RESULTS: Through the stimulation of hCD40-mCD27 molecule via anti-human CD40 antibody or CD154 ligation, expression of CD11a, CD23, CD54, CD70 and CD80 were increased and secretion of IgM was induced, which were comparable to the effect of CD40 stimulation. TRAF2 and TRAF3 were recruited into lipid-enriched membrane raft and were bound to CD27 in M12.4.1 cells. CD27 stimulation, however, did not increase TRAF2 or TRAF3 degradation. CONCLUSION: In contrast to CD40 signaling pathway, TRAF2 and TRAF3 degradation was not observed after CD27 stimulation and it might contribute to prolonged B cell activation through CD27 signaling.

Simultaneous Detection of Adenovirus types 3, 7 and 11 by a Multiplex PCR from Cell Cultured Clinical Specimens of Hospitalized Patients

Adenovirus (AD) is classified into A, B, C, D, E and F subgenera on the basis of neutralization, hemagglutination and DNA homology tests. Six AD subgenera have 51 serotypes. We selected AD types 3, 7, and 11 since they are known to frequently cause respiratory disease, gastrointestinal disease and meningitis, which often require hospitalization of the patients. The purpose of this study was to detect and to evaluate the correlation of AD types 3, 7 and 11 and the diseases. The detection method, used for AD types 3, 7 and 11, was a multiplex polymerase chain reaction (PCR) with HEp-2 cell cultured with clinical samples. The result was as follows: AD 3 (22/62, 35.5%), AD 7 (27/62, 43.5%), and AD 11 (50/62, 80.6%) were detected by the multiplex PCR in 62 clinical samples. AD 3 detection rate in cerebrospinal fluid was relatively higher than in other specimens but its statistical significance was low (p=0.07). AD 11 was detected (50/62, 80.6%) with the highest frequency and appeared to significantly be associated with gastroenteritis, respiratory disease, and meningitis in hospitalized children, and arthritis in adult patients. AD 3, AD 7 and AD 11 types were detected singly in 21/62 (33.9%), doubly in 15/62 (24.2%), and triply in 16/62 (25.8%). The multiplex PCR method using Hep2 cell culture with clinical specimens could detect AD types 3, 7, and 11 in children with meningitis, respiratory infection, or gastroenteritis, and in adults with cancer and arthritis.

Detection of Adenovirus and Parvovirus in CSF Specimens from Viral Meningitis Adult Patients by Enzyme Immunoassay with Monoclonal Antibody and by Nested - PCR

Viral meningitis and encephalitis are important and serious diseases in young children and adults. There are many causative viruses but it is known that a low percentage of adenovirus (ADV) and parvovirus (PA V) infected individuals develop meningitis or encephalitis. Few reports have been published about central nervous system complications that were rare but fatal. First we used enzyme immunoassay (EIA) with monoclonal antibody to detect ADV antigen (Ag) and PAV Ag in cerebrospinal fluids (CSF) from acute phase of hospitalized adult patients with viral meningitis or viral encephalitis. Second we detected ADV DNA and PAV DNA in the same CSF after cell culture by nested-polymerase chain reaction (PCR). Third we evaluated ADV and PAV dual infection in CSF by EIA and nested-PCR. ADV Ag in CSF by EIA positivity was 42.9% (12l28) and PAV Ag in CSF by EIA positivity was 21.4% (6/28). ADV DNA in CSF by nested-PCR positivity was 89.3% (25/28) and PAV DNA in CSF by nested-PCR positivity was 38.5% (10/26). ADV and PAV dual infection in CSF by 11CSted-PCR was 35.7% (10/28). Detection rate of ADV DNA and PAV DNA in CSF by nested-PCR with viral meningitis or encephalitis adult patients were higher than we expected. Positive detection of nested-PCR was higher than that of EIA with monoclonal antibody for detection of antigens ADV and PAV in CSF with viral meningitis or encephalitis adult patients. Both methods were analnized by the McNemar test.

Haemophilus influenzae and Human Herpes Virus-6 Detection by EIA and PCR in CSF from Young Hospitalized Children with Aseptic Meningitis

There are various microbial agents which causing meningitis in children. It is well known that Haemophilus influenzae (HI) are the most frequent bacterial agents. In Korea, it is hard to find studies detecting HI in cerebrospinal fluid (CSF) from pediatric patients with aseptic meningitis by PCR. It has been also reported that human herpes virus-6 (HHV-6) causes meningitis, meningoencephalitis, neuroinvasion and persistent infection of the central nervous system in children. In this study we also detected HHV-6 in the same CSF by EIA and PCR. We used 85 CSF specimens from 85 aseptic meningitis patients (mean age 6.6 years) taken from the Department of pediatrics at Ewha Womans University MokDong Hospital from June 2001 to August 2002. Detection rate of HI by EIA method was 12.9% (11/85) and by PCR was 16.5% (14/85). Detection rate of HHV-6 by EIA and by PCR was 18.8% (16/85) and 21.2% (18/85), respectively. Co-detection rate of HI and HHV-6 was 7.1% (6/85) by EIA and 12.9% (11/85) by PCR. In conclusion, by PCR in combination with EIA, HI infection could be proved in the aseptic meningitis CSF from which no bacterium was cultivated.

Detection by PCR of Adenovirus and Human Herpes Virus 6 in Peripheral Blood Monocyte from Young Children who were Hospitalized with Lower Respiratory Tract Infection

There are reports that the second most causative viral agent which causes lower respiratory tract infection (LRTI) in young children is adenovirus (ADV). Human herpes virus 6 (HHV-6) is also reported as a rare agent of LRTI in young children. But there is no report of simultaneous detection of ADV and HHV-6 in LRTI using the same peripheral blood monocyte (PBM) by nested-polymerase chain reaction (PCR) or PCR. Firstly, we detected ADV antigen (Ag) and HHV-6 Ag in serum by each monoclonal antibody with enzyme immunoassay (EIA). Secondly we tested two viruses in peripheral blood monocyte by nested-PCR or PCR. Twenty nine cases of young hospitalized children with LRTI (mean age 11.3 months, mean hospitalization period 5.7 days) had bronchiolitis or viral pneumonia and were confirmed by X-ray findings. Positivity of ADV Ag in serum by EIA was 75% (21/28) and positivity of HHV-6 Ag in serum by EIA was 10.7% (3/28). ADV in PBM by nested-PCR positivity was 89.7% (26/29) and HHV-6 in PBM by PCR positivity was 42.9% (12/28). ADV and HHV-6 dual infection in PBM by PCR was 11/29 (37.9%). Young children with dual infection were hospitalized (mean 6.3 days) with severe bronchiolitis.

Detection of Streptococcus pneumoniae Pneumolysin Gene by PCR in Sera and Cerebrospinal Fluids from Hospitalized Patients

No abstract available.

Molecular Mechanism of TNF-alpha and MMP-9 Production in Response to HIV-1 Core Antigen p24 in Human Monocytie THP-1 Cells

No abstract available.

Detection of Herpes Virus-6 by PCR I Cerebrospinal Fluid from Hospitalized Adult Patients with Aseptic Meningitis or Encephalitis

No Abstract Available.

Detection of Coinfection and Persistent Infection of Adenovirus and Varicella-Zoster Virus in Synovial Fluids From Synovitis Patients by Nested-PCR

No Abstract Available.

Interaction of HIV-1 core p24 antigen with human monocytic cell line THP1 results in TNF-alpha dependent secretion of matrix metalloproteinase-9

Immunological mechanisms involving the release of inflammatory factors by HIV-1 infected microglia in the brain have been implicated in the pathogenesis of HIV dementia (HIVD). Since the regulation of matrix metalloproteinases (MMPs) activity can be influenced by variety of inflammatory mediators, this study was undertaken to look for a correlation between the MMP-9 release and the production of TNF-alpha in response to HIV-1 p24 in the human monocyte cell line THP-1 as a model for microglia. First, it was shown that HIV-1 core p24 antigen induced THP-1 to secrete MMP-9 in a dose response manner while it elicited a little effect on MMP-2 release in human astroglial cell line T98G. Next, it was found that p24 induced THP-1 to secrete TNF-alpha without prior differentiation into macrophages by phorbol myristate acetate (PMA) treatment. Furthermore, anti-TNF-alpha neutralizing antibodies significantly blocked p24-induced MMP-9 release in a dose dependent manner. Our data indicate that p24 antigen induces monocytic MMP-9 release by triggering up-regulation of TNF-alpha secretion.

Detection and differentiation of Herpes Simplex Virus 1 and 2, and Varicella-Zoster virus in vesicle fluid, joint fluid and serum using PCR method

The viruses of Herpes Simplex Virus 1 (HSV-1), Herpes Simplex Virus 2 (HSV-2) and Varicella-Zoster virus (VZV) which belong to the alpha herpes subfamily are important human pathogens. When eruptions were not fully developed from these viral infections, clinical diagnosis was not always easy and required virological confirmation test. The above viruses were reactivated in individuals who were compromised in immune competence for one reason or another. Polymerase chain reaction (PCR) enables rapid and sensitive detection of HSV and VZV DNAs. Its sensitivity was largely influenced by choice of primers. Authors conducted a study to detect of those three viruses in human specimens including vesicle fluid and joint fluid and serum using PCR methods. Primers used for this study were the general primer pair GPHV-RU which was known to amplify within the genes enjoying the highest degree of homology between UL15 of HSV and UL42 of VZV. PCR with primers hybridized pair GPHV-RU amplifies a 396 bp with HSV-1 and HSV-2 standard stain DNA and 405 bp with VZV standard strain DNA. Restriction enzyme cleavage with HpaII and DdeI were used to detect and distinguish DNAs of HSV-1 and HSV-2 and VZV. The purpose of this study was a rapid and easy detection of VZV and HSV-1 or HSV-2 from various clinical specimens (vesicle fluid, serum and joint fluid) by PCR method. Used methods were: HSV PCR with primer 1, 2 and HpaII RE digestion; VZV nested PCR; HSV PCR with primer A, B and BssHII RE digestion. 1) In 33 cases (33/42, 78.6%) VZV was detected single or mixed infection from 42 clinical specimens which included vesicle fluid (5), serum from respiratory infected children (10), serum from immune suppressed adult cancer patients (7) and joint fluid from arthritis patients (20). 2) In 20 cases (20/42, 42.6%) HSV was detected singly or mixed infection and 19 of the cases were HSV-2 and 1 case was HSV-1. 3) In 19 cases (19/42, 45.2%) VZV was singly detected which included serum from respiratory infected children (6 cases), joint fluid from arthritis patients (9 cases), vesicle fluid (2 cases) and serum form immunosuppressed cancer patients (2 cases). 4) HSV was singly detected in 6 cases (6/42, 14.3%) which included joint fluid from arthritis patients (5 cases) and serum form respiratory infected children (1 cases). 5) 14 cases of VZV and HSV mixed infection (14/42, 33.3%) were detected. They included vesicle fluid (3 cases), serum form immunosuppressed cancer patients (4 cases), serum from respiratory infected children (2 cases) and joint fluid from arthritis patients (5 cases). 6) HSV-1 and HSV-2 detection and typing by HSV PCR with primer A, B and BssHII RE digestion method was more sensitive and the results were easier to detect than on other method.

Detection of Astrovirus Infection from Hospitalized Young Children Feces by Reverse Transcription - Polymerase Chain Reaction

Astrovirus is frequently associated with diarrhea in children. It can not be readily isolated by cell culture, and an electronmicroscope is usually used for detection of this agent. Recently in 1995 a combined method of reverse transcription-polymerase chain reaction (RT-PCR) was designed for easier detection of astrovirus, which is based on the conserved sequence in 3'-end of genomes of the 7 known serotypes of human astrovirus. As of yet there has not been any report of astrovirus data in Korea using the RT-PCR methods. The purpose of this study was to detect astrovirus incidence, severity of symptoms, seasonal variation and coinfection rate with rotavirus in Korean children inpatients with diarrhea. Fecal specimens from 61 young children hospitalized with gasteroenteritis Korea from Jan. 1996 through Mar. 1997. They were examined for astroviurs infection by RT-PCR method. Results are as follows: 1. Astrovirus was detected at 9.8% (6/61) from fecal specimens of children with severe diarrhea by EIA using monoclonal antibody coated plates. 2. Astorvirus was detected at 29.5% (18/61) from fecal specimens of children with severe diarrhea by RT-PCR. 3. The age of the 18 children affected by astrovirus ranged from 2 monthes to 7 years with mean of 3.0 years. 4. Mean hospital stay of the 1S children was 6.1 days. 5. Five (27.8%) astrovirus RT-PCR positive strains were confirmed in November and in December, respectively out of 18 specimens in total. 6. Astrovirus coinfection with rotavirus type G1 was confirmed in 15/16 specimens (93.8%), and with type G2 was in 1/16 specimens (6.3%).

Measurement of Apoptosis by Staining with 7-Amino-Actinomycin D with Concurrent Dual Color Immunofluorescence by Single Laser Flow Cytometry

Apoptosis is a crucial mechanism for the selective elimination of mammalian cells and is involved in many physiological and pathological processes. The heightened awareness of the importance of apoptosis has increased the need for rapid and quantitative methods for measurement of apoptotic changes. Recently, 7-amino-actinomycin D (7-AAD) has been introduced as a valuable fluorescent dye for assessing apoptosis by flow cytometry. When the cells are stained with 7-AAD in the concentration of 10 - 20 ug/ml, live cells are not stained (7-AAD ) and early apoptotic cells are weakly stained (7-AAD ) while late apoptotic or dead cells are stained brightly (7-AAD). On scattergram of forward angle light scatter vs. 7-AAD fluorescence, the three populations can be discriminated not only between each other but also from cell debris or clumps. The apoptotic cells, defined as 7- AAD cells, were demonstrated as apoptotic by morphological observation of the sorted cells. The 7-AAD cell fraction was also demonstrated to be parallel with subdiploid fraction of cells stained with PL However, 7-AAD cells, whose definition is based on the alteration of membrane integrity, have never been demonstrated to be subdiploid fraction by simultaneous DNA staining. Here we directly demonstrate that 7-AAD cells, defined on the scattergram of forward angle light scatter vs. 7-AAD fluorescence, are subdiploid fraction by staining with DNA dye whose fluorescence is collected after 530/30 band pass filter (FL-1). We also demonstrate the effects of 7-AAD concentration, fixation of cells, and proliferation of cells, on the fluorescence pattern, for reference during assessment of apoptosis by simple and rapid method for flow cytometric analysis of cells stained with 7- AAD. We also present a flow cytometric analysis of cells stained with 7-AAD Eor sequential change in apoptotic fraction, with concurrent dual-color immunophenotyping.

VP7 Genotypes of Human Rotavirus from Hospitalized Children with Severe Diarrhea by Reverse Transcription - Polymerase Chain Reaction

Human rotavirus has now been established as the leading cause of gastroenteritis in young children worldwide. At least fourteen serotypes of group A rotavirus have been identified on the basis of antibody responses to major neutralizing glycoprotein, VP7 (G type for glycoprotein), present in the outer capsid of the virus. Serotype 1, 2, 3 and 4 are the most highly prevalent in human. In Korea, rotavirus is also the principal cause of severe nonbacterial diarrhea requiring hospitalization in infants and young children, which is commonly detected by EIA method. The epidemiology of rotavirus infection has been monitored by only serologic methods without electropherotyping in Korea. This study shows seasonal and age related variations .of rotavirus infection in Korea according to the genotype using the reverse transcription polymerase chain reaction (RT-PCR). Fecal specimens were obtained from 39 children hospitalized with acute watery diarrhea and gastroenteritis in Ewha Womans University MokDong Hospital in Seoul from Jan. to Dec. of 1996. All four (1, 2, 3, 4) major G serotypes were identified by amplification of segment of the gene for VP7 using RT-PCR. Rotavirus Gl 749 bp, G2 653 bp, G3 374 bp and G4 583bp were shown on 2.9 or 3.3% NuSieve agar gel. Results were as follows: 1) Rotavirus was detected at 53.8% (21/39) by EIA and 89.7% (35/39) by RT-PCR. 2) Serotype Gl, G2, G3, G4 when detected by RT-PCR accounted for 80.0% (28/35), 14.3% (5/35), 2.9% (1/35) and 2.9% (1/35), respectively. 3) Thirty five strains of rotavirus were detected at the frequency of 17.1% (6/35) in Oct., 20.0% (7/35) in Nov. and 20.0% (7/35) in Dec. 4) As for the age range, children affected by rotavirus were mostly under 1 years.

The Expression of Fcgamma Receptors in Cord and Adult Blood Granulocytes

PURPOSE: The immature neonatal immune system is thought to result in increased risk of infection. Receptors for the Fc moiety of IgG (FcgammaR) are important in antibody-mediated clearance of microbes by granulocytes and monocytes/macrophages. As an approach to understand their role in neonatal life, we compared the constitutive expression of the three Fc receptors-FcgammaRI (CD64), FcgammaRII (CD32) and FcgammaRIII (CD16)-of neonatal granulocytes with those of adult granulocytes. METHODS: Using quantitative immunofluorescence by flow cytometry, we have compared the constitutive expression of the three Fc receptors-FcgammaRI (CD64), FcgammaRII (CD32) and FcgammaRIII (CD16)-by neonatal and adult blood granulocytes. RESULTS: The proportion of FcgammaRI+ granulocytes is significanly high in cord blood compared to that of adult. However, the proportions of FcgammaRII+ or FcgammaRIII+ cells are significanly low in cord blood compared to those of adlut. CONCLUSIONS: There were differences in the expression of FcgammaR on granulocytes between cord blood and adult peripheral blood. This results suggest that the differences in FcgammaR expression may be contributed to the differences in functional activity.