Polymorphisms in the promoter regions of the CXCL1 and CXCL2 genes contribute to increased risk of alopecia areata in the Korean population.

Alopecia areata (AA) is a common disease, which causes hair loss in humans. AA has a genetically complex inheritance. This study investigated the possible correlations between single nucleotide polymorphisms (SNPs) in the promoter regions of the chemokine (C-X-C motif) ligand 1 (melanoma growth stimulating activity, alpha) (CXCL1) and chemokine (C-X-C motif) ligand 2 (CXCL2) genes and the development of AA in the Korean population. Two hundred and thirty-five AA patients and 240 control subjects were recruited. The specific SNPs occurring in the promoter regions of the CXCL1 and CXCL2 genes (rs3117604, -429C/T and rs3806792, -264T/C, respectively) were genotyped. All data obtained was evaluated using the SNPStats, SPSS 18.0, and the Haploview v.4.2 software platforms. The Odd's ratios (OR), 95% confidence intervals (CI), and P values were calculated using multiple logistic regression models. Analyses of the genetic sequences obtained revealed a significant correlation between the two SNPs and the development of AA (rs3117604, P = 0.0009 in co-dominant model 1, P = 0.01 in co-dominant model 2, P = 0.004 in the dominant model, P = 0.005 in the log-additive model, P = 0.012 in allele distribution; rs3806792, P = 0.036 in co-dominant model 2, P = 0.0046 in the log-additive model). The TT and CC haplotypes were also observed to show a significant association with increased risk of AA (TT haplotype, P = 0.0018; CC haplotype, P = 0.0349). Our data suggests that the CXCL1 and CXCL2 genes may be associated with AA susceptibility.

Pancytopenia and secondary myelofibrosis could be induced by primary hyperparathyroidism.

Hyperparathyroidism may be a precipitating factor important to the development of myelofibrosis: however, there has been only a few reports regarding myelofibrosis secondary to primary hyperparathyroidism. Recently, a rare case of pancytopenia caused by myelofibrosis in a 41-year-old woman who complained of general weakness and arthralgia presented to our clinical service. The patient was diagnosed with primary hyperparathyroidism with pancytopenia. Bone marrow biopsy revealed myelofibrosis. Right parathyroidectomy was performed and a parathyroid adenoma was totally excised. After surgery, the CBC counts and other clinical abnormalities gradually improved without further intervention. We concluded that the pancytopenia was because of bone marrow fibrosis resulting from primary hyperparathyroidism. Therefore, physicians should consider myelofibrosis secondary to primary hyperparathyroidism as a cause of pancytopenia in hypercalcemic patients, even though it is rare.

Long term results from percutaneous radiofrequency neurotomy on posterior primary ramus in patients with chronic low back pain.

PURPOSE: We report on our experience of percutaneous radiofrequency neurotomy (PRN) on the posterior primary ramus (PPR) with at least two years follow-up. METHODS: 228 patients underwent PRN on the PPR for refractory low back pain. 128 patients met the inclusion criteria of facetal originated pain (group II), while 100 patients did not (group I). Radiofrequency (RF) procedures were applied in the usual manner. Pain relief was assessed at 1 week, 1 month, 6 months and 2 years using visual analog scale (VAS). FINDINGS: Positive responders were 56% at 1 week, 46% at 1 month, 18% at 6 months, and 13% at 2 years after PRN in group I, and 78.9% at 1 week, 75.4% at 1 month, 62.5% at 6 months, and 54.7% at 2 years in group II. Prominent local tenderness, percussion tenderness, pain on getting up, extension and transitional movement, radiating pain on buttock and/or posterior thigh, and good immediate response were found to be significantly related to good outcome. CONCLUSION: PRN on the PPR has long-term beneficial effects. Long-term good results can be achieved after proper selection of patients with facet joint related low back pain.

A new and simple transection knife for study of neurodegeneration and neuroregeneration in animal model.

BACKGROUND: The purpose of this study was to design and make a simple, inexpensive brain knife that could produce consistent results following transection in animal model. MATERIALS AND METHODS: After testing various materials including commercially available products, microelectrode recording needles as used in deep brain stimulation (DBS) surgery were selected as ideal candidates. They were modified to serve as type of wire-knife for the purposes of study. For this study, the major pathway for dopaminergic neuron from substantia nigra to striatum was selected for transection. A total of 40 Sprague-Dawley rats were assigned to 8 groups; normal, 1-4, 6, 8, and 10 weeks post-transection. Degree of cell death was determined and surviving neurons were counted by means of fluorescent microscopic examination, immunohistochemistry involving tyrosine hydroxylase (TH)-immunoreactive staining, and mapping to verify complete transection. RESULTS: Compared to control, percentage of remaining neurons in each group was 61.3, 36.8, 29.9, 5.1, 5.9, 7.0%, respectively. Completeness of lesion was correlated with the absence of TH-immunoreactivity in the striatum. CONCLUSION: Our model seems to provide complete cell death in early period after transection with consistent results. Thus, this type of brain knife can be very handy, without any extra cost, in any research model involving transection of fiber bundle for studies on neurodegeneration and neuroregeneration.

Knowledge of and attitude towards circumcision of adult Korean males by age.

AIM: Circumcision is widely practised in Korea, but little is known regarding the public's attitude towards circumcision. This study was designed to evaluate the knowledge and the general opinion of Korean adult males towards circumcision. METHODS: Fifteen hundred self-completion questionnaires were distributed to adult males in five decadal age groups ranging from 10 to 59 y old. Questions concerning opinions regarding the necessity, reasons, potential benefits and disadvantages of circumcision, as well as the role of peer pressure upon the decision to circumcise were included. Completed questionnaires were collected and analysed statistically. RESULTS: The achieved response rate was 62.7%. 73.1% believed that circumcision is necessary, while 7.1% believed it is not necessary. The principal reason for circumcision was to improve penile hygiene (77.9%). 68.7% did not prefer neonatal circumcision regardless of the respondent's age. The major reason was fear of pain (36.9%). Peer pressure was one of the most influential factors when deciding upon circumcision: 60.8% believed that they might be ridiculed by their peer group unless circumcised, and the younger the age of the respondent, the more frequently this opinion was held (p < 0.05). 62.7% thought that circumcision would prevent genital tract infection of the sexual partner. Respondents with older age tended to emphasize improved sexual potency (p < 0.05). CONCLUSIONS: This study indicates that common beliefs of adult males about circumcision in Korea are relatively homogeneous. Tailored education about circumcision is needed.

Nocturia in patients with lower urinary tract symptoms: association with diurnal voiding patterns.

OBJECTIVE: To determine whether diurnal voiding patterns predict nocturia in patients with lower urinary tract symptoms (LUTS), as few studies have evaluated the association between diurnal and nocturnal voiding patterns. PATIENTS AND METHODS: We prospectively analysed the frequency-volume charts (FVCs) of consecutive patients with LUTS. At the initial visit patients had a detailed clinical evaluation and subsequently were requested to complete a 72-h FVC. In all, 104 (41 men and 63 women, mean age 63 years, range 50-83) were included in the primary analyses. Associations between daytime variables and nocturia were described using maximum likelihood estimates of the relative risk and by 95% confidence intervals (CIs) based on logistic regression models. RESULTS: When at least one night-time void was used to define nocturia the multivariate logistic model showed a negative association of mean daytime voided volume with nocturia (P = 0.001). The odds ratio for nocturia decreased with this variable to 0.98 (95% CI 0.96-0.99). When 'voiding at least twice per night' was used to define nocturia only the number of daytime voids was positively related to nocturia (odds ratio 1.22; 95% CI 1.01-1.48; P= 0.040). CONCLUSION: Nocturia may be associated with diurnal voiding patterns; these results also suggest that the causes of nocturia of one or of two or more voids may differ. This highlights the role of bladder function in more severe forms of nocturia.

Ultralow anterior resection and coloanal anastomosis for distal rectal cancer: functional and oncological results.

We evaluated the functional and oncological outcome of ultralow anterior resection and coloanal anastomosis (CAA), which is a popular technique for preserving anal sphincter in patients with distal rectal cancer. Forty-eight patients were followed up for 6-100 months regarding fecal or gas incontinence, frequency of bowel movement, and local or systemic recurrence. The main operative techniques were total mesorectal excision with autonomic nerve preservation; the type of anastomosis was straight CAA, performed by the perianal hand sewn method in 38 cases and by the double-stapled method in 10. Postoperative complications included transient urinary retention (n=7), anastomotic stenosis (n=3), anastomotic leakage (n=3), rectovaginal fistula (n=2), and cancer positive margin (n=1; patient refused reoperation). Overall there were recurrences in seven patients (14.5%): one local and one systemic recurrence in stage B2; and one local, two systemic, and two combined local and systemic in C2. The mean frequency of bowel movements was 6.1 per day after 3 months, 4.4 after 1 year, and 3.1 after 2 years. The Kirwan grade for fecal incontinence was 2.7 after 3 months, 1.8 after 1 year, and 1.5 after 2 years. With careful selection of patients and good operative technique, CAA can be performed safely in distal rectal cancer. Normal continence and acceptable frequency of bowel movements can be obtained within 1 year after operation without compromising the rate of local recurrence.

Activation of NF-kappa B by nontypeable Hemophilus influenzae is mediated by toll-like receptor 2-TAK1-dependent NIK-IKK alpha /beta-I kappa B alpha and MKK3/6-p38 MAP kinase signaling pathways in epithelial cells.

Nontypeable Hemophilus influenzae (NTHi) is an important human pathogen in both children and adults. In children, it causes otitis media, the most common childhood infection and the leading cause of conductive hearing loss in the United States. In adults, it causes lower respiratory tract infections in the setting of chronic obstructive pulmonary disease, the fourth leading cause of death in the United States. The molecular mechanisms underlying the pathogenesis of NTHi-induced infections remain undefined, but they may involve activation of NF-kappa B, a transcriptional activator of multiple host defense genes involved in immune and inflammatory responses. Here, we show that NTHi strongly activates NF-kappa B in human epithelial cells via two distinct signaling pathways, NF-kappa B translocation-dependent and -independent pathways. The NF-kappa B translocation-dependent pathway involves activation of NF-kappa B inducing kinase (NIK)--IKK alpha/beta complex leading to I kappa B alpha phosphorylation and degradation, whereas the NF-kappa B translocation-independent pathway involves activation of MKK3/6--p38 mitogen-activated protein (MAP) kinase pathway. Bifurcation of NTHi-induced NIK-IKK alpha/beta-I kappa B alpha and MKK3/6--p38 MAP kinase pathways may occur at transforming growth factor-beta activated kinase 1 (TAK1). Furthermore, we show that toll-like receptor 2 (TLR2) is required for NTHi-induced NF-kappa B activation. In addition, several key inflammatory mediators including IL-1 beta, IL-8, and tumor necrosis factor-alpha are up-regulated by NTHi. Finally, P6, a 16-kDa lipoprotein highly conserved in the outer membrane of all NTHi and H. influenzae type b strains, appears to also activate NF-kappa B via similar signaling pathways. Taken together, our results demonstrate that NTHi activates NF-kappa B via TLR2-TAK1-dependent NIK--IKK alpha/beta-I kappa B alpha and MKK3/6--p38 MAP kinase signaling pathways. These studies may bring new insights into molecular pathogenesis of NTHi-induced infections and open up new therapeutic targets for these diseases.

Distal anterior cerebral artery aneurysms: clinical features and surgical outcome.

Aneurysms of the distal anterior cerebral artery (DACA) are rare and their surgical treatments present some unique difficulties from a technical standpoint. In this report, we presented our experiences of cases with DACA aneurysms, and analyzed the clinical features and prognostic factors affecting the final outcomes. Among 770 cases of intracranial aneurysms operated from 1990 to 1998, 19 cases of DACA aneurysms (2.5%) were studied retrospectively. The characteristic findings were female preponderance (M:F = 1:2.8), common multiple aneurysms (57.9%), and frequent intracerebral hemorrhage (ICH) on initial brain CT scan (42.1%). All patients were operated via interhemispheric approach. Intraoperative aneurysmal rupture was developed only in 3 cases (15.8%), and had no relationship with the final outcome Fifteen out of 19 patients (78.9%) showed favorable outcome with a mortality rate of 5.3%. The follow-up data suggest that the initial ICH on brain CT scan portend a poor prognosis.

Immortalization of rat middle ear epithelial cells by adeno 12-SV40 hybrid virus.

Rat middle ear epithelial cells were infected with the adeno 12-SV40 hybrid virus. The cell line thus obtained displays features of primary cultured epithelial cells in both light microscopic and ultrastructural examinations. The immortalized cells have been in continuous proliferation for 40 passages and more than 17 months. Immunohistochemical analysis of the immortalized cells was positive for the SV40 T antigen and the tumor suppressor protein p53. The cells also stained positive for cytokeratin, an epithelial cell marker, and negative for vimentin, a fibroblast marker. These results, together with karyotype analysis, indicate that this cell line originated from rat middle ear epithelial cells and retains the characteristics of epithelial cells. This cell line will be useful for studying the normal cellular biology of middle ear epithelial cells, as well as the cellular and molecular mechanisms involved in the bacteria-middle ear epithelial cell interaction.

Chronic subdural hematoma: evaluation of the clinical significance of postoperative drainage volume.

OBJECT: A wide variation in postoperative drainage volumes is observed during treatment of chronic subdural hematoma (CSDH) with twist-drill or burr-hole craniostomy and closed-system drainage. In this study the authors investigate the causes of the variation, the clinical significance thereof, and its influence on treatment outcome. METHODS: A total of 175 cases were investigated between January 1991 and December 1997. Of these, 145 patients had surgery for CSDH, of whom 30 had bilateral lesions. The cases of CSDH were divided into five subtypes (low-density, isodense, high-density, mixed-density, and layering types) on the basis of the brain computerized tomography (CT) findings. Burr-hole craniostomies with closed-system drainage were performed in all patients and the drainage was maintained for 5 days, during which daily amounts of fluid were measured. The mean drainage volume over 5 days was 320 ml, with the largest volume (413 ml) seen in the low-density type and the smallest (151 ml) in the mixed-density type of CSDH. There were recurrences in six patients (seven instances, 4%). The mixed-density type had the highest recurrence rate (8.6%), whereas there was no recurrence for the low-density type. There were no recurrences in 81 patients in whom the total drainage volumes for 5 days were more than 200 ml, but there were recurrences in six (seven instances) of 94 patients in whom the total drainage volume was less than 200 ml. CONCLUSIONS: The postoperative drainage volumes varied greatly because of differences in the outer membrane permeability of CSDH, and such variation seems to be related to the findings on the CT scans obtained preoperatively. Patients with CSDH in whom there is less postoperative drainage than expected should be carefully observed, with special attention paid to the possibility of recurrence.

Cell biology of tubotympanum in relation to pathogenesis of otitis media - a review.

The sterility of the eustachian tube and tympanic cavity of normal individuals is maintained not only by the adaptive immune system, but also by the mucociliary system and the antimicrobial molecules of innate immunity. Mucin production and periciliary fluid homeostasis are essential for normal mucociliary function and dysfunction of this system is an important risk factor for otitis media. The secreted antimicrobial molecules of the tubotympanum include lysozyme, lactoferrin, beta defensins, and the surfactant proteins A and D (SP-A, SP-D). Defects in the expression or regulation of these molecules may also be the major risk factor for otitis media.

Mucin gene expression in cultured human middle ear epithelial cells.

For the advanced study of the cell and molecular biology of middle ear mucosa, an in vitro cell culture system is required. Although middle ear epithelial cells have been cultured from various species of laboratory animal, there have been no reports concerning a serial subculture system of human middle ear epithelial cells. In this paper, we describe the establishment of a primary culture system of human middle ear epithelial cells using a serum-free conditioned medium and the characterization of these cells by the expression of phenotypic characteristics of epithelial cells and mucin genes. Cultured cells were anchorage-dependent in terms of growth and showed a polygonal cobblestone-like appearance: desmosomes in the cell junction were observed by electron microscopy. In the immunocytochemical study, cytokeratin (epithelial cell marker) was expressed in all cultured cells. but von Willebrand factor (endothelial cell marker) was not. Unexpectedly, vimentin (fibroblast marker) was locally expressed, and a double stain showed the co-expression of both cytokeratin and vimentin in the same cell. The products of reverse transcriptase polymerase chain reaction from cultured cells yielded distinct bands compatible with the expected sizes of the MUC1, MUC2, MUC5AC and MUC5B genes. This culture system will allow us to prepare the cell line and to perform advanced studies of human middle ear mucosal biology.

Effects of retinoic acid, triiodothyronine and hydrocortisone on mucin and lysozyme expression in cultured human middle ear epithelial cells.

Mucous hypersecretion is a major complication of otitis media and can prolong the disease course and increase morbidity. Mucin, a major component of mucus, is a macromolecular complex of glycoprotein and makes mucus viscous. Lysozyme is a secretory element of the middle ear mucosa. which has a non-specific and innate antibacterial function. We attempted to identify factors that regulate these secretory products and their morphological phenotype using cultured human middle ear epithelial cells. Cellular differentiation was induced by creating an air liquid interface on culture day 9 in serum-free conditioned media. Omission of retinoic acid (RA) caused decrease in the secretion of mucin and lysozyme, and in the cellular expression of MUC 2, MUC 5AC and MUC 5B mRNA. In contrast, removal of triiodothyronine (T3) caused an increase in the secretion of mucin and the level of MUC5AC mRNA. When hydrocortisone (HC) was removed from the media, the secretion of mucin was decreased with out an apparent change of message level. The expression of MUC 1 mRNA was not changed by the respective deficiency of RA. T3 or HC. The effect of T3 or HC on lysozyme was not significant. This study shows that RA, T3 and HC influence the morphological phenotype and the secretory function of mucin and lysozyme in cultured human middle ear epithelial cells. This culture system can serve as an in vitro model for study of the regulation of various cellular secretions in human middle ear epithelium.

Immortalization of chinchilla middle ear epithelial cells by adenovirus 12-simian virus 40 hybrid virus.

In order to study the cellular and molecular mechanisms of the pathogenesis of otitis media, a chinchilla middle ear epithelial cell line (CMEE-1) with differentiated cell characteristics was established by infection of a primary culture with the adenovirus 12-simian virus 40 (Ad12-SV40) hybrid. This cell line has been in continuous culture for 42 passages, whereas the parent cells underwent senescence and died at the 8th passage. The cell line also retains epithelial morphology and expresses cytokeratin polypeptides 4, 7, and 18, characteristic markers for epithelia. In Western blots of cell proteins, bands at 94 and 53 kd were labeled after binding antibodies against SV40 large T antigen and p53, respectively. Karyotype analysis showed that the cell line is derived from chinchilla epithelial cells. These findings confirm that the cell line is a chinchilla epithelial cell immortalized by the hybrid virus.

Coexpression of neurotrophic growth factors and their receptors in human facial motor neurons.

Neuronal development and maintenance of facial motor neurons is believed to be regulated by neurotrophic growth factors. Using celloidin-embedded sections, we evaluated immunoreactivity of 11 neurotrophic factors and their receptors in facial nuclei of human brain stems (4 normal cases, and 1 from a patient with facial palsy and synkinesis). In the normal subjects, positive immunoreactivity of the growth factor neurotrophin-4 and acidic fibroblast growth factor (aFGF) was observed in facial motor neurons, as was positive immunoreactivity against ret, the receptor shared by glial cell line-derived neurotrophic factor and neurturin. Immunoreactivity was moderate for the receptor trkB and strong for trkC. In the case of partial facial palsy, surviving cells failed to show immunoreactivity against neurotrophins. However, immunoreactivity of aFGF was up-regulated in both neuronal and non-neuronal cells in this patient. Results suggest that these trophic growth factors and their receptors may protect facial neurons from secondary degeneration and promote regrowth of the facial nerve after axotomy or injury.

Identification of common lipooligosaccharide types in isolates from patients with otitis media by monoclonal antibodies against nontypeable Haemophilus influenzae 9274.

Twenty-one murine monoclonal antibodies (MAbs) were induced by nontypeable Haemophilus influenzae (NTHi) 9274. Nineteen MAbs were specific for the lipooligosaccharide (LOS) as determined by enzyme-linked immunosorbent assay (ELISA) and Western blot analysis. When the MAbs were assayed with five LOS prototype strains by ELISA, all bound to strain 3198 LOS (type III), while six of the MAbs were also reactive with LOSs from strain 1479 (type I), 5657 (type IV), or 7502 (type V). Ten MAbs had complement-mediated bactericidal activity, and three MAbs were opsonophagocytic against the homologous strain. Five LOS MAbs with different specificities were used to analyze 155 NTHi clinical isolates from the United States and from Japan. These isolates were classified into nine groups by ELISA. Only four isolates (2.6%) were not recognized by any of the five MAbs. Most of the isolates (91.6%) were in four groups which bound three of the five MAbs. One of three MAbs, 6347C11, had strong activity against the homologous strain and was also bactericidal to 45 clinical isolates (29%) which belonged to the four common patterns (25 belonged to pattern 1). These data indicate that these MAbs can be used for LOS typing in which almost all NTHi strains can be typed according to the LOS antigenicity. Among NTHi, at least one conserved LOS epitope which is a target of bactericidal antibodies exists. We conclude that strain 9274 LOS, which is the target for bactericidal antibodies, is a candidate for LOS-based NTHi vaccines.

Effects of bisphosphonate on experimental jaw metastasis model in nude mice.

The mechanism of osteolysis associated with metastatic cancer of the jaws is essentially osteoclast-mediated. Therefore, it is likely that potent osteoclastic bone resorption inhibitors such as bisphosphonates would be efficacious for the treatment of jaw metastasis. We examined the effects of a third generation bisphosphonate, YM175, in a nude mice jaw metastasis model with intracardiac injection of a human breast cancer cell line, MDA-MB-231. The metastatic lesions in untreated mice were radiographically observed at the body and angle of the mandible. Histology of the mandible of untreated mice revealed that most of the bone marrow cavities had been occupied by the metastatic tumor with active osteoclasts along the trabecular bone. The experimental group showed that YM175 markedly reduced the size of tumor and the number of osteoclasts. These results suggest that YM175 may suppress metastasis formation and tumor growth in jaw through inhibition of osteoclastic bone resorption.