Bacille Calmette Guérin (BCG) can induce Kawasaki disease-like features in programmed death-1 (PD-1) gene knockout mice.

OBJECTIVES: Various genetic variants of inhibitory immune signals have been suspected as feasible causes of Kawasaki disease (KD). We investigated the associative role of programmed death-1 (PD-1) gene in the pathogenesis of KD by injecting bacilli Calmette Guérin (BCG) to PD-1 gene knockout (PD-1KO) mice. METHODS: In order to induce KD-like clinical manifestations in young PD-1KO mice, intradermal injection of the bacilli Calmette Guérin (BCG) was performed twice on the abdominal skin with a 4-week interval. For defining the role of BCG, heat shock protein (HSP) 65 was challenged. In addition, Staphylococcus aureus was adopted as a microorganism that does not contain HSP65 structure. One month after the second injection, heart, liver, and kidneys were removed and examined. RESULTS: PD-1KO mice showed KD-like features including prolonged fever for more than 5 days, erythematous swelling on soles, tail skin desquamation, and gallbladder (GB) hydrops. Inflammatory cell aggregation and intimal proliferation in at least more than one coronary artery was found in all PD-1KO mice whereas scanty coronary lesion was found in wild type (WT) mice. When the PD-1KO mice were injected twice with HSP65, coronary arterial lesions similar to those seen after BCG injection were observed. Inflammatory reactions in other organs including hepatic arteries, renal arteries, and biliary arteries were also observed in PD-1KO mice. CONCLUSIONS: Our data suggest that PD-1 gene may be one of the genetic predispositions of KD and antigens containing HSP65 structure could be a triggering factor of KD by our animal model of KD.

Establishing a surveillance network for severe lower respiratory tract infections in Korean infants and young children.

To reduce morbidity and mortality through integrated case management, a pilot study to detect respiratory viruses in patients with acute lower respiratory infections (ALRIs) was designed as part of a nationwide surveillance for this disease in Korea. The study population consisted of hospitalized patients under the age of 5 years with bronchiolitis, pneumonia, croup, or acute respiratory distress syndrome. A prospective 6-month study was performed. Two hundred and ninety-seven nasopharyngeal secretions were collected and multiplex reverse transcriptase polymerase chain reactions (RT-PCR)/polymerase chain reactions (PCR) were performed to detect respiratory viruses. If there were any positive RT-PCR/PCR results, viral cultures were proceeded for confirmation. Respiratory viruses were identified in 49.6% of 296 patients. The detection rates were as follows: respiratory syncytial virus (RSV) was the most commonly detected in 52.7% (87/165), human metapneumovirus (hMPV) in 15.8%, human corona virus (hCoV) in 5.5%, adenovirus in 9.7%, human bocavirus (hBoV) in 5.5%, parainfluenza virus (PIV) in 3.6%, rhinovirus (RV) in 4.2%, and the influenza virus in 3% of the patients with ALRIs. The consistent rate of positive results between RT-PCR and viral culture was 92% (105/114). Using our methods to detect viral causes seemed to be acceptable for the national surveillance of severe acute respiratory infections in infants and children.

Prevalence and patterns of anti-nuclear antibodies in Korean children with juvenile idiopathic arthritis according to ILAR criteria.

OBJECTIVES: To investigate the prevalence and patterns of anti-nuclear antibodies (ANA) in different subtypes of juvenile idiopathic arthritis (JIA) according to the International League of Associations for Rheumatology (ILAR) criteria. METHODS: One hundred and fifty-three Korean patients (M:F 83:70) with JIA were followed between 1990 and 2006 and were tested for ANA by an indirect immunofluorescence method using HEp-2 cells as the substrate. ANA tests were repeated in 37 patients during the course of the disease. The median age at onset was 7.5 years (range 0.8-15.9 years). RESULTS: ANA were positive in 50 (33%) of the 153 patients at a dilution of 1:40 or higher (>1:40 in 70%, >1:80 in 2%, >1:160 in 16%, >1:320 in 2%, and >1:640 in 10%). The patterns of immunofluorescence staining were homogeneous in 50%, speckled in 38%, nucleolar in 8%, and centromere in 4%. ANA titres were decreased in 25 (68%) of the 37 patients, and the nuclear fluorescence patterns changed in 14 (38%) during follow-up. ANA seropositivity was associated with female sex (p<0.0001), negative HLA-B27 (p = 0.01), and a persistently elevated erythrocyte sedimentation rate (ESR) at follow-up (p = 0.014). Furthermore, a high ANA titre (>1:160) was associated with a poor clinical outcome (active patients at follow-up) (p = 0.005). CONCLUSIONS: ANA may be an important marker of disease activity in patients with JIA. ANA titres tend to decrease during disease remission but the fluorescence patterns do not appear to be related to disease activity or clinical outcome.

Elevated anti-alpha-enolase antibody levels in Kawasaki disease.

OBJECTIVE: By functioning as a heat-shock protein (HSP), alpha-enolase has an important role in the pathophysiology of multivariant vasculitis. Kawasaki disease (KD) is a type of vasculitis occurring primarily in children. The role of alpha-enolase in KD was assessed by measuring anti-alpha-enolase antibody (Ab) titres in patients with KD and the usefulness of anti-alpha-enolase Ab as a diagnostic tool in atypical KD patients was evaluated. METHODS: Anti-alpha-enolase Ab titres were measured by using an enzyme-linked immunosorbent assay (ELISA) in seven normal control patients, nine febrile control patients and 14 KD patients (10 typical KD, four atypical KD). A standard deviation (SD) of 3 above the mean of the normal control group was considered to be positive reactivity. Western blotting using recombinant human alpha-enolase was performed in four KD patients and three normal controls. RESULTS: With the positive reactivity limited to +3 SD over the mean (>0.6), 10 out of 14 patients (71%) were positive at the acute onset and 12 out of 14 patients (85.7%) were positive before discharge. In total, 12 out of 14 patients (85.7%) were positive either at acute onset or before discharge. All four atypical KD patients showed positive reactivity. Specific positive bands against recombinant human alpha-enolase were detected by western blotting in all four KD patients, but no reactivity was seen in three patients with normal controls. CONCLUSION: This is the first study to demonstrate that autoantibodies against the alpha-enolase are present in the sera of KD patients. We suggest that anti-alpha-enolase Ab should be a good candidate for a diagnostic tool in atypical KD.

Effect of pregnancy on food consumption and consciousness factors associated with food satisfaction.

To examine the effect of pregnancy on food consumption, we surveyed the recall of past experience of two groups of women, those who had been pregnant (PY, n=188) and those who never had (PN, n=111), regarding their food consumption and related dietary behaviors. The questionnaire, answered with regard to potato chips, contained 135 objective components expressing sensorial stimuli and 103 subjective consciousness components, including knowledge, education, faith, memory, experience, lifestyle, family values, imagination, and mental state. In the PY group, 86% of the women change in food consumption and preference during pregnancy, and 60% experienced decreased food consumption during emesis gravidarum (EG). The change in food consumption during and after periods of EG was influenced by the number of previous births for the women in the PY group; in women pregnant for a second or third time, the change in food intake was less than during the previous pregnancy. The difference in food satisfaction with regard to potato chips between the PY and PN groups showed that overall food satisfaction could not be explained by a combination of objective sensorial components; the only objective components that were directly related to overall satisfaction were taste and texture. Multidimensional analysis with strength of sensorial stimulus, preference, and overall satisfaction revealed differences in patterns between the PN and PY groups. The effect of the consciousness components on food preference and satisfaction was comparable to that of the sensorial components.

Increased serum levels of macrophage migration inhibitory factor in patients with Kawasaki disease.

OBJECTIVE: To determine whether serum levels of macrophage migration inhibitory factor (MIF) increase in patients with Kawasaki disease (KD) and also correlate with other inflammatory indices. METHODS: Serum samples from 10 patients with KD, 15 normal healthy subjects, and seven febrile control subjects were assayed for MIF by enzyme-linked immunosorbent assay (ELISA). RESULTS: There was a significant increase in the serum levels of MIF in the acute stage of KD [113.06 (range 20.6-157.36) ng/mL] compared with those in the subacute stage [28.11 (8.57-143.48) ng/mL, p<0.01], normal controls [12.95 (8.40-18.67) ng/mL, p<0.001], and febrile controls [36.58 (21.31-59.67) ng/mL, p = 0.01]. The increase in MIF correlated with an increase in interleukin-6 (IL-6) (r = 0.52, p = 0.047). CONCLUSION: MIF may be a useful marker in the acute stage of KD and may provide important clues to the pathogenesis of this disease.

[DNA methylation on urinalysis and as a prognostic marker in urothelial cancer of the bladder]. / DNA-Methylierung in der Urindiagnostik und als Prognosemarker beim Urothelkarzinom der Harnblase.

INTRODUCTION AND OBJECTIVES: Detection of promoter hypermethylation has been proposed as a promising tool for cancer diagnosis and as a prognostic marker in various cancers. We studied the versatility of DNA methylation for noninvasive diagnosis and as a prognostic marker for non-muscle-invasive bladder carcinoma. METHODS: Tumor specimens were microdissected and DNA was extracted from 105 paraffin-embedded paraffin specimens from patients undergoing transurethral resection for non-muscle-invasive bladder carcinoma. Urine specimens were collected from patients undergoing cystectomy for bladder cancer and from healthy volunteers. Methylation status was assessed with the real-time quantitative methylation-sensitive PCR (MethyLight). We checked a panel of 20 cancer-associated genes (p14ARF, p16 CDKN2A, STAT-1, SOCS-1, DR-3, DR-6, PIG-7, BCL-2, H-TERT, BAX, EDNRB, DAPK, RASSF-1A, FADD, TMS-1, E-CADHERIN, ICAM-1, TIMP-3, MLH-1, COX-2) for DNA methylation. RESULTS: Follow-up data were available in 95 of 105 patients (91.4%). A tumor recurrence was observed in 26 patients (27.3%). We could identify six genes (SOCS-1, STAT-1, BCL-2, DAPK, TIMP-3, E-cadherin), where methylation was associated with tumor recurrence. In Kaplan-Meier analysis, TIMP-3 showed a significant association with recurrence-free survival. Methylation of TIMP-3 predicted prolonged disease-free interval. Regarding urinalysis we could identify a pattern of methylation markers including DAPK, BCL-2, and H-TERT that yielded a sensitivity of 81.1% with a specificity of 100% in a cancer-free control population CONCLUSIONS: We present data on the clinical usefulness of methylation analysis in bladder carcinoma. Our data confirm that methylation analysis is a promising tool for bladder cancer diagnosis and prognosis.

Intraosseous hemangioma of the zygoma: CT and MR findings.

Intraosseous hemangiomas are uncommon, constituting less than 1% of all osseous tumors. The most frequent sites are the calvaria and the vertebral column. Involvement of the facial bones is rare, and occurs most commonly in the maxilla, mandible, and nasal bones. Only 20 cases of zygomatic involvement have been reported in the English-language literature. We report a case of an intraosseous hemangioma of the zygoma documented by CT and MR studies.

Latissimus dorsi musculocutaneous flap based on the serratus branch with microvascular venous augmentation.

The authors report a case of a latissimus dorsi musculocutaneous flap with microvascular venous supercharging in the setting of an acute ligation of the thoracodorsal vascular pedicle. Through their experience with this case, they have questioned a common belief that the flap will survive without the thoracodorsal vessels intact. They accept that risk is minimized when rotation of the flap takes place several weeks after ligation of the vessels. However, they do not believe it is prudent to transfer a latissimus flap of significant size immediately after ligation of the thoracodorsal vessels, unless the vascular supply is enhanced.

Radial forearm free flap for salvage of Achilles-tendon repair wounds.

Achilles-tendon repair wounds are prone to breakdowns and secondary infections, compromising the integrity of the repair and the ultimate function of the repaired tendon. Unlike defects resulting from tumor excision, burns, or trauma, Achilles-tendon repair wounds have their own special considerations. The dissection needed for the repair of the tendon often denudes the tendon of the peritenon blood supply and creates a sub-tendinous dead space. Anatomic topography unique to the Achilles region makes the choice of flap coverage crucial in terms of function and cosmesis. Considering all the flap options utilized in the past, the radial forearm free flap is the most logical choice, offering the advantage of thin, supple, vascularized tissue. The flap can also be elevated easily for secondary reconstructions, when simultaneous tendon reconstruction and coverage are not possible. The authors present their experience utilizing the radial forearm flap as the flap of choice for salvage of the complicated wounds of Achilles tendon repair.

Salvage of ear replantation using the temporoparietal fascia flap.

The authors report a case involving a 46-year-old man who sustained a traumatic amputation of approximately 60% of his ear from a human bite. The ear was replanted microsurgically without the benefit of venous anastomosis. Blood transfusion was not required despite the use of leech therapy and systemic anticoagulation. The replantation appeared to be a success at the time of his discharge from the hospital, but during the late postoperative period the replanted ear became progressively necrotic. The failing ear replantation was rescued successfully with the use of temporoparietal fascia flap reconstruction.

Staging arteriovenous fistula loops for lengthening of free-flap pedicles.

The authors investigated the optimal period of maturation following the creation of arteriovenous (AV) loops using polyterafluoroethylene (PTFE) in a white rat model, which were subsequently used to support free-tissue transfer The AV loops in Group 1 (n = 17) were allowed to mature for 3 days prior to creation of the flap, while those from Group 2 (n = 14) were allowed to mature for 5 days. Results were compared to those from a previous study in which the authors reported an 80 percent initial patency rate (n = 30) and a 67 percent viability rate, based on 12 patent loops after 7 days. In the present study, patency rates were 59 percent for the 3-day group and 79 percent for the 5-day group; viability rates were 50 and 64 percent, respectively. Considering both patent and nonpatent loops, the overall viability rates were 29 and 50 percent respectively. Maturation periods longer than 3 days for AV loops constructed from PTFE micrografts were determined to be preferable for subsequent free-tissue transfer.

Power of politics and reasonableness in policy study: on some methodological problems with the Harvard Team Report.

The so-called "Harvard Team Report," commissioned by the Hong Kong government (Hong Kong SAR Government, 1999), suggests significant institutional changes to the local health care system, including a partial shift of the financial burden directly to the citizens. I argue that 1) the Report's adoption of the contextuality principle as its research framework encounters practical problems in collecting data for a reliable analysis; 2) the existing health care system already satisfies the Report's first guiding principle; 3) the Report's employment of the "working assumption" of the government (i.e., not increasing its financial support of health care) as its second guiding principle is questionable, for the share of the percentage of GDP as represented by the existing system (4.6% in 1996) is small enough; and 4) because of 3), the Report is unnecessarily constrained in its choices of considered options and seems to overlook some feasible ones. In conclusion, the methodological reasonableness of the Report is questioned.

Use of the umbilicus in reconstruction of the vulva and vagina with a rectus abdominis musculocutaneous flap.

Flap reconstruction of the vulva and vagina following gynecological ablative procedures has become an integral part of the management of gynecological oncology patients. The benefits of flap reconstruction, including early primary healing, improved cosmesis over skin grafting, and prolonged secondary wound healing, have been well accepted. Additionally, the creation of a neovagina or neovulva often restores the sexual function and positive body image of the patient lost to radical procedures. The gracilis musculocutaneous flap has been used extensively in flap reconstruction but reports of partial flap necrosis and the need for extensive dissection of both thighs have led to alternative flap choices. The rectus musculocutaneous flap, with its hardier cutaneous blood supply, is often too bulky and difficult to inset delicately around the preserved urethral and vaginal cuff. The umbilicus has the required soft tissue, and its conically contoured depression allows for delicate insetting of the rectus abdominis musculocutaneous flap around the urethral cuff. We present 2 patients who underwent vertical rectus musculocutaneous flaps with umbilical soft tissue to restore urinary function and to create a cosmetic nonfunctional vagina.

Characterization of guanidine transport in human renal brush border membranes.

PURPOSE: Organic cation transporters in the renal proximal tubule are important in the secretion of many clinically used drugs and their metabolites. The goal of this study was to determine the mechanisms of guanidine transport in human kidney. METHODS: Brush-border membrane vesicles were prepared from donor human kidneys deemed unsuitable for renal transplantation. RESULTS: Uptake of [14C]-guanidine (50 microM) in the vesicles, as determined by rapid filtration, was significantly greater in the presence of an outwardly-directed proton gradient, at all early time points, than in the absence of the gradient. Proton-stimulated uptake of [14C]-guanidine at 30 sec (32.0 +/- 1.24 pmol/mg protein) was significantly inhibited by a number of organic cations including 5 mM unlabeled guanidine (14.8 +/- 1.84 pmol/mg protein) and 5 mM MIBA (9.14 +/- 3.80 pmol/ mg protein), but not by 5 mM TEA (28.4 +/- 5.67 pmol/mg protein). Guanidine, but not TEA, trans-stimulated [14C]-guanidine uptake. Conversely, TEA, but not guanidine, trans-stimulated [14C]-TEA uptake in the vesicles. The proton-dependent transport of guanidine was characterized by a Km of 3.52 +/- 0.42 mM (SE) and a Vmax of 34.6 +/- 8.64 pmol/mg protein/sec (SE). CONCLUSIONS: These results demonstrate that guanidine transport in human renal brush border membrane vesicles is stimulated by a proton gradient. Evidence was obtained suggesting that the transporter for guanidine is distinct from the previously described organic cation proton antiporter for TEA.

Cloning and functional characterization of a rat renal organic cation transporter isoform (rOCT1A).

Polyspecific organic cation transporters in the renal proximal tubule mediate the secretion of many clinically used drugs as well as endogenous metabolites. Recently, two organic cation transporters (rOCT1 and rOCT2) were cloned from rat kidney. In this study, we report the cloning and functional expression of an rOCT1 isoform, rOCT1A, from rat kidney. Genomic DNA cloning and sequencing demonstrated that rOCT1A is an alternatively spliced variant of rOCT1 with a deletion of 104 base pairs near the 5'-end. The uptake of [14C]tetraethylammonium (TEA) in oocytes injected with the cRNA-encoding rOCT1A was increased 16-fold over that in water-injected oocytes (29 +/- 2.8 pmol/oocyte/h versus 1.8 +/- 0.13 pmol/oocyte/h, mean +/- S.E., p < 0.05). [14C]TEA uptake in the cRNA-injected oocytes was saturable (Km = 42 +/- 11 microM) and was inhibited significantly by organic cations, including cimetidine and N1-methylnicotinamide. The amino acid sequence was deduced from the cDNA after examination of all three reading frames. Two overlapping open reading frames were found. Studies with synthetic constructs suggest that a functional organic cation transporter is encoded by the larger open reading frame. The larger open reading frame encodes a 430-amino acid protein (termed rOCT1A) that is 92% identical to rOCT1 and 57% identical to rOCT2. From hydropathy analysis, rOCT1A is predicted to have 10 transmembrane domains with both amino and carboxyl termini intracellular. RNase protection assays demonstrate the presence of rOCT1A mRNA transcripts in rat kidney cortex, medulla, and intestine. These studies demonstrate the presence of a functional, alternatively spliced organic cation transporter (rOCT1A) in rat kidney.

Expression of human polyspecific renal organic cation transport activity in Xenopus laevis oocytes.

Polyspecific organic cation transporters in the basolateral and brush border membrane of the kidney play a role in the elimination of many clinically important drugs and endogenous compounds. In this study we report the functional expression of organic cation transport activity in Xenopus laevis oocytes injected with poly(A)+RNA (mRNA) isolated from human kidney. Uptake of [14C]tetraethylammonium (TEA) was measured in mRNA-injected or water-injected oocytes, 4 days after injection. In oocytes injected with 50 ng of mRNA isolated from human renal cortex, the uptake of [14C]TEA was significantly increased in comparison with water-injected oocytes (7.2 +/- 0.6 and 3.5 +/- 0.3 pmol/oocyte/h, respectively). Injection of 20 ng of an enriched size-fraction (fraction C) of mRNA (mean size of 2.3 kb) resulted in further enhancement of [14C]TEA uptake: [14C]TEA uptake was enhanced six-to seven-fold in oocytes injected with fraction C (23.7 +/- 3.7 pmol/oocyte/h) in comparison with water-injected oocytes. The uptake of TEA in mRNA-injected oocytes was significantly inhibited by 5 mM of unlabeled TEA, cimetidine, and N1-methylnicotinamide. These data suggest that polyspecific organic cation transport activity can be successfully expressed in Xenopus laevis oocytes injected with mRNA isolated from human kidney.

Increased patency of artificial microvascular grafts using arteriovenous fistula loops: a two-stage procedure for lengthening the pedicle of free-tissue transfer.

The effect of a high-flow system on the patency of artificial microvascular grafts (AMG) was investigated using arteriovenous (A-V) fistula loops made of polytetrafluoroethylene (PTFE) in a clinically useful longer length. At a second stage, the A-V loops, matured for 7 days, were used as recipient conduits for free-tissue transfer. The patency rates of 5-cm long PTFE A-V fistula loops were compared with 5-cm and 1-cm long interpositional micrografts in a rat model. The patency rate of the loops at 7 days was 80 percent, compared to 0 percent for both the 5-cm and 1-cm interpositional grafts. A-V fistula loops matured for 7 days in vivo were divided in their midsections and used as arterial and venous conduits (each 2.5 cm long) for free epigastric flaps (3 x 6 cm) raised from the contralateral groin. Sixty-seven percent (8/12) of the free flaps were viable at 7 days. The A-V fistula loop in a high-flow system has a beneficial effect in maintaining patency of AMGs. PTFE fistula loops matured for 7 days can be used as arterial and venous conduits for microsurgical tissue transfer in rats.

Microsurgical reconstruction of difficult orocutaneous fistulas.

We present 5 cases from a series of eight orocutaneous fistulas that were successfully reconstructed through the use of free tissue transfer. Reconstruction was performed in 5 patients with a radial forearm flap and in 3 patients with a free jejunal transfer. This report addresses the approach to reconstructing the difficult orocutaneous fistula with emphasis on free flap design and the use of a bilaminar flap.