Supplementary Material for: Natural Killer Cell Deficits Aggravate Allergic Rhinosinusitis in a Murine Model

<b><i>Objective:</i></b> Defective innate immune functions can contribute to chronic rhinosinusitis (RS). Recently, it has been reported that chronic RS patients show impaired function of natural killer (NK) cells. We investigated the role of NK cells in eosinophilic inflammation in an allergic RS mouse model. <b><i>Methods:</i></b> Mice sensitized to ovalbumin (OVA) by intraperitoneal injection received nasal challenges with OVA for 5 weeks. NK cell depletion was achieved by intraperitoneal injections of anti-asialo ganglio-N-tetraosylceramide (ASGM1) antibodies 10 days before OVA sensitization and every 5 days thereafter until sacrifice. Sinonasal complex samples were evaluated histologically, and IL-4, IL-5, IL-13, IFN-γ, MIP-2, and eotaxin levels were measured in the nasal lavage fluid. Differential white blood cell counts were also obtained. <b><i>Results:</i></b> Allergic RS mice showed significantly more eosinophilic inflammation in the sinonasal mucosa, elevated levels of IL-4, IL-5, IL-13, and eotaxin in the nasal lavage fluid, and peripheral blood eosinophilia compared to control mice. The depletion of NK cells by anti-ASGM1 treatment induced more prominent eosinophilic inflammation and increased secretion of IL-5 and peripheral blood eosinophilia in allergic RS mice. <b><i>Conclusion:</i></b> The depletion of NK cells aggravates allergen-induced sinonasal eosinophilic inflammation, suggesting that impaired NK cell activity may be an exacerbating factor in eosinophilic chronic RS

Supplementary Material for: Survival in Subcortical Vascular Dementia: Predictors and Comparison to Probable Alzheimer's Disease in a Tertiary Memory Clinic Population

<b><i>Background:</i></b> Subcortical vascular dementia (SVaD) is one of the most common dementias, after Alzheimer's disease (AD) dementia. Few survival analyses in SVaD patients have been reported. <b><i>Methods:</i></b> The dates and causes of death of 146 SVaD and 725 AD patients were included. We used the Cox proportional hazards model to compare survival between SVaD and AD patients and to explore possible factors related to survival of SVaD patients. <b><i>Results:</i></b> The median survival time after the onset of SVaD (109 months) was shorter than that recorded for AD (152 months). The most common cause of death in SVaD was stroke (47.1%). Factors associated with shorter survival in SVaD were late onset, male sex, worse baseline cognition, absence of hypertension and a family history of stroke. <b><i>Conclusions:</i></b> Stroke prevention may be important in SVaD treatment because 47.1% of SVaD patients died of stroke. A family history of stroke and absence of hypertension were associated with a shorter survival in SVaD, suggesting the existence of genetic or unknown risk factors

Supplementary Material for: Gemcitabine plus Cisplatin versus Capecitabine plus Cisplatin as First-Line Chemotherapy for Advanced Biliary Tract Cancer: A Retrospective Cohort Study

<b><i>Background and Aim:</i></b> Gemcitabine plus cisplatin (GP) is a standard chemotherapy option for patients with advanced biliary tract cancer (BTC). We compared the efficacy and safety of capecitabine plus cisplatin (XP) versus GP in advanced BTC. <b><i>Methods:</i></b> The records of all patients treated with GP or XP chemotherapy for unresectable, metastatic, or recurrent BTC at the National Cancer Center between December 2001 and August 2012 were reviewed retrospectively. Patients with histologically confirmed intrahepatic cholangiocarcinoma, gallbladder cancer, or extrahepatic cholangiocarcinoma were enrolled. <b><i>Results:</i></b> Of the 344 patients enrolled, 127 received GP and 217 received XP. At a median follow-up time of 8.9 months, the median time to progression was longer in the GP group than in the XP group (5.6 vs. 4.7 months), but the difference was not statistically significant (p = 0.081). The median overall survival (OS) was 8.4 months (95% CI 6.2-10.7) in the GP group and 7.6 months (95% CI 6.8-8.7) in the XP group (p = 0.024), with statistical significance retained following multivariate analysis (HR 0.72; 95% CI 0.527-0.987; p = 0.004). Grade 3/4 toxicities were significantly more frequent in the GP group than in the XP group (40.9 vs. 24.9%, p = 0.002). <b><i>Conclusions:</i></b> GP was superior to XP in prolonging OS, despite increasing the rate of grade 3/4 adverse events

Erratum: Clinical and Endocrine Features of Two Allan-Herndon-Dudley Syndrome Patients with Monocarboxylate Transporter 8 Mutations

The monocarboxylate transporter 8 <i>(MCT8)</i> gene, located on chromosome Xq13.2, encodes a thyroid hormone transporter that is involved in triiodothyronine (T3) uptake into central neurons. <i>MCT8</i> mutations cause an X-linked syndromic disorder known as Allan-Herndon-Dudley syndrome (AHDS) that is characterized by severe psychomotor delays, abnormal thyroid function, and hypomyelinated leukodystrophies. We identified 2 AHDS patients with developmental delays, truncal hypotonia, and spastic paraplegia. These patients presented with psychomotor retardation and characteristic thyroid function abnormalities, such as elevated T3 and low T4 levels. Direct <i>MCT8</i> sequencing identified heterozygous mutations in each patient: p.I114N and p.A224V, respectively. Because it is difficult to suspect AHDS solely according to neurological features, thyroid function, including the T3 level, should be screened in male patients with X-linked mental retardation. Although the clinical features of hypothyroidism cannot be improved by only administering levothyroxine treatment, early diagnosis, management, and appropriate genetic counseling should be provided to at-risk families

Supplementary Material for: Free Fatty Acid Is Associated with Thrombogenicity in Cardioembolic Stroke

<i>Background:</i> Recently, the role of free fatty acids (FFAs) in thromboembolism has re-emerged in the context of cardioembolic stroke. Therefore, we attempted to determine the role of FFAs in embolic risk in various potential sources of cardioembolism (PSCE). We hypothesized that if elevated FFA levels in stroke patients are associated with thrombogenesis, then patients with a well-known high risk of embolic sources would have high FFA levels. <i>Methods:</i> Data collected from 2 hospital-based stroke registries were analyzed to investigate the association between FFA and PSCE. <i>Results:</i> A total of 2,770 acute stroke patients, including 539 with cardioembolic stroke, were selected for analysis. FFA was an independent predictor for cardioembolism (OR 2.755, 95% CI 2.221-3.417, <i>p</i> < 0.001). Among the PSCE, FFA levels were significantly associated with high risk of atrial fibrillation (AF), valvular heart disease, congestive heart failure with low ejection fraction, left atrial thrombus, left ventricular thrombus, left atrial smoke, and ventricular wall motion abnormality. FFA levels increased with the number of PSCE per patient without interaction with the presence of AF. <i>Conclusions:</i> Among acute stroke patients, FFA levels increased in groups with higher risk of cardioembolic stroke irrespective of the presence of AF. These results suggest that enhanced thrombogenicity could be the main mechanism to explain the elevated FFA levels in patients with cardioembolic stroke

Supplementary Material for: Novel Heterozygous Mutations of <b><i>NR5A1</i></b> and Their Functional Characteristics in Patients with 46,XY Disorders of Sex Development without Adrenal Insufficiency

<b><i>Background/Aims:</i></b> Heterozygous mutations of <i>NR5A1</i>, which encodes steroidogenic factor 1 (SF1), were identified in patients with 46,XY disorders of sex development (DSD) with normal adrenal function. This study was aimed to identify and functionally characterize mutations of <i>NR5A1</i> in patients with 46,XY DSD. <b><i>Methods:</i></b> This study included 51 patients from 49 unrelated families with 46,XY DSD. Genomic DNA was extracted from peripheral blood leukocytes, and direct sequencing of all coding exons and their flanking introns of <i>NR5A1</i> was performed. Transient transfections and dual-luciferase® reporter assays were performed to evaluate the effect of <i>NR5A1</i> variants on transcriptional activity. <b><i>Results:</i></b> Four of 49 patients (8.2%) harbored a novel heterozygous sequence variant of <i>NR5A1</i>: c.80G>C (p.G26A), c.847T>C (p.C283R), c.1151del (p.L384Rfs*7), and c.1333G>T (p.E445*). They presented with female external genitalia with clitoromegaly in infancy or childhood, or primary amenorrhea in adolescence. In vitro functional studies of SF1 activity determined that each variant, except p.E445*, led to a reduced expression of downstream target genes and disturbed the regulation of gonadal development. <b><i>Conclusions:</i></b> Loss-of-function mutations of <i>NR5A1</i> are a relatively common cause of 46,XY DSD. Therefore, genetic defects of <i>NR5A1</i> should be considered as an etiology in subjects with 46,XY DSD without adrenal insufficiency

Supplementary Material for: Experimental Inhibition of Periostin Attenuates Kidney Fibrosis

<b><i>Background:</i></b> Periostin is responsible for tissue regeneration, fibrosis, and wound healing via its interaction with integrin. Recently, the role of periostin has been shown to contribute to fibrosis in chronic kidney disease. We investigated the role of periostin and the effect of periostin blockade in renal fibrogenesis. <b><i>Methods:</i></b> We investigated the function of periostin in vivo in wild-type and periostin-null mice (Postn-KO) in a unilateral ureteral obstruction (UUO) model. For the in vitro experiments, primary cultured inner medullary collecting duct cells from the wild-type and Postn-KO mice were used. <b><i>Results:</i></b> Periostin expression was strongly induced by UUO in the wild-type mice. UUO induced renal fibrosis and morphological changes in the obstructed kidney of wild-type mice, whereas global knockout of periostin reduced fibrosis induced by UUO and improved kidney structure. Fibrosis- and inflammation-related mRNA were significantly induced in the wild-type mice and were decreased in the Postn-KO mice. Additionally, α-smooth muscle actin expression was increased following the administration of recombinant periostin in vitro. The effect of periostin blockade was examined using 2 methods. The integrin blockade peptide decreased fibrosis-related gene expression in in vitro experiments. Anti-periostin polyclonal antibody attenuated renal fibrosis induced by UUO through changes in transforming growth factor-β signaling and the inflammatory and apoptotic pathways. <b><i>Conclusion:</i></b> Periostin is a marker of renal fibrosis and may augment the progression of fibrogenesis as an extracellular matrix protein. Periostin blockade effectively attenuated renal fibrogenesis. Thus, periostin inhibition may be a therapeutic strategy for the amelioration of renal disease progression

Supplementary Material for: Detection of Human Papillomavirus DNA by DNA Chip in Breast Carcinomas of Korean Women

<p>It remains unclear whether there is an association between human papillomavirus (HPV) infection and human breast cancer. The aim of this study was to investigate the presence of HPV DNA in breast carcinomas of Korean women and to examine the possible association between HPV and breast cancer development. For this purpose, HPV DNAs from 154 patients, including 123 patients with breast carcinoma and 31 with intraductal papilloma, and nipple tissue from 27 cancer patients were examined using the DNA chip method. HPV DNA was detected in 8 breast carcinomas (6.5%) but in no intraductal papilloma. All detected HPV genotypes were of high-risk groups. There was a slightly increased incidence in papillary carcinomas (11.5%) and invasive ductal carcinomas with adjacent intraductal papillomas (11.8%) compared to the other histological subtypes (3.2–4.3%), although the difference was not statistically significant (p = 0.126). The presence of HPV DNA was not correlated with specific prognostic predictors of disease. High-risk HPV DNA sequences were present in 6.5% of Korean patients with breast tumors. However, this study could not demonstrate whether or not such HPVs directly contribute to the development of breast cancer.</p

Supplementary Material for: Age-Dependent Predictors for Recurrent Stroke: The Paradoxical Role of Triglycerides

<b><i>Background:</i></b> Although long-term predictors of mortality and vascular events after ischemic stroke are well defined, the age-dependent differences in predicting outcomes are unknown, particularly for lipid parameters. <b><i>Methods:</i></b> We assessed recurrent stroke and other vascular events in patients with first-ever ischemic strokes who were registered in a prospectively collected hospital-based stroke registry. Patients were classified into a middle-age group (40–64 years old) and an old-age group (65 years and older). <b><i>Results:</i></b> A total of 551 patients comprising 235 middle-age and 316 old-age subjects were investigated. At the mean follow-up of 26.4 months, 49 (8.9%) patients had experienced recurrent stroke. Outcome events in the middle-age group (3.8%) were less frequent than in the old-age group (12.7%). The effects of vascular risk factors were different across age groups. In particular, the hazard ratio direction for triglycerides was significantly different between age groups for recurrent stroke or composite outcomes. <b><i>Conclusions:</i></b> Incidence and predictors of stroke and other vascular events following ischemic stroke are age-dependent. Differential effects of triglycerides on long-term outcomes following ischemic stroke were found to be hazardous at middle age but turned out to be non-significant at old age

Supplementary Material for: Urinary Periostin Excretion Predicts Renal Outcome in IgA Nephropathy

<i>Background:</i> Periostin is a matricellular protein and plays a vital role in tissue regeneration, fibrosis and wound healing. However, data about its significance in nephrology are limited. We investigated the correlation between urinary periostin excretion and its clinical significance including renal histologic findings and prognosis in IgA nephropathy (IgAN). <i>Methods:</i> Of 399 patients from a glomerulonephritis cohort recruited between January 2009 and December 2014, 314 were enrolled. Serum and urine periostin (uPOSTN) were measured using enzyme-linked immunosorbent assay. We divided the patients into 3 groups by uPOSTN/creatinine (uPOSTN/Cr): group 1 (undetectable), group 2 (lower than the median) and group 3 (higher than the median). <i>Results:</i> The uPOSTN level was correlated with pathologic classifications and both initial and final IDMS-MDRD estimated glomerular filtration rates (eGFRs; p < 0.001). Histologically, group 3 patients were correlated with severe interstitial fibrosis/tubular atrophy (p = 0.004), interstitial inflammation (p = 0.007), hyaline arteriolosclerosis (p = 0.001) and glomerular sclerosis (p < 0.001). A higher initial uPOSTN/Cr level was associated with a greater decline in eGFR during follow-up (p = 0.043 when initial eGFR ≥60; p = 0.025 when eGFR <60 ml/min/1.73 m2), and the renal outcomes with end-stage renal disease (ESRD; p = 0.003), ESRD and/or eGFR decrease of >30% (p = 0.033) and ESRD and/or eGFR decrease of >50% (p = 0.046) occurred significantly more in group 3. In multivariate analysis, uPOSTN group 3 (hazards ratio 2.839, 95% CI 1.013-7.957; p = 0.047) was independently associated with ESRD in IgAN patients. <i>Conclusion:</i> uPOSTN/Cr value at initial diagnosis correlated with renal fibrosis and predicted the renal outcomes in patients with IgAN. It could be a promising urinary biomarker for renal fibrosis