Moonkyu Kim

Glypican‐3 is overexpressed in human hepatocellular carcinoma

To identify candidate genes that could be used as diagnostic and therapeutic targets for hepatocellular carcinoma (HCC), we searched for the genes that are overexpressed in HCC by combining representational difference analysis and microarray. Genes such as glypican‐3 (GPC3), insulin‐like growth factor 2, long‐chain fatty‐acid‐coenzyme A ligase 4, farnesyl diphosphate synthase were frequently identified in our screening. Northern blot analysis with these four genes confirmed their overexpression in HCC. Among them we found that GPC3 transcript is upregulated in six out of seven cases of HCC. Immunoblot and immunohistochemical staining using polyclonal anti‐GPC3 antibodies further confirmed that GPC3 protein is indeed increased in HCC tumor samples. We also found that GPC3 is secreted into culture media from cell lines derived from HCC. We conclude that GPC3 is a good molecular marker for HCC. (Cancer Sci 2003; 94: 259–262)

A Guide to Studying Human Hair Follicle Cycling In Vivo

Hair follicles (HFs) undergo life-long cyclical transformations, progressing through stages of rapid growth (anagen), regression (catagen), and relative “quiescence” (telogen). Since HF cycling abnormalities underlie many human hair growth disorders, the accurate classification of individual cycle stages within skin biopsies is clinically important and essential for hair research. For preclinical human hair research purposes, human scalp skin can be xenografted onto immunocompromised mice to study human HF cycling and manipulate long-lasting anagen in vivo. While available for mice, a comprehensive guide on how to recognize different human hair cycle stages in vivo is lacking. Here, we present such a guide, which uses objective, well-defined, and reproducible criteria and integrates simple morphological indicators with advanced, (immuno)-histochemical markers. This guide also characterizes human HF cycling in xenografts and highlights the utility of this model for in vivo hair research. Detailed schematic drawings and representative micrographs provide examples of how best to identify human HF stages, even in sub-optimally sectioned tissue, and practical recommendations are given for designing human-on-mouse hair cycle experiments. Thus, this guide seeks to offer a benchmark for human hair cycle stage classification, for both hair research experts and newcomers to the field.

Enhanced transfollicular delivery of adriamycin with a liposome and iontophoresis.

Abstract: To find a better way to deliver drugs into hair follicles, we tried two approaches: single topical application using various liposomes; and iontophoresis combined with topical application of ionic liposome. After delivery of adriamycin (ADR) to wax‐depilated rat skin, the transport of the drug was examined under fluorescence microscopy.

Fatty acid‐CoA ligase 4 is overexpressed in human hepatocellular carcinoma

Fatty acid‐CoA ligase 4 (FACL4) is a central enzyme controlling the unesterified arachidonic acid (AA) level in cells. It has been shown that FACL4 blocks apoptosis and promotes colon carcinogenesis by lowering the cellular level of unesterified AA. Consistent with this, FACL4 is upregulated in colon adenocarcinoma. The status of FACL4 in other tumors including hepatocellular carcinoma (HCC) is not known. Here, we report that FACL4 is overexpressed in human HCC compared with adjacent normal liver tissues. FACL4 mRNA and protein were overexpressed in 5 out of 12 (41.7%) and 3 out of 8 (37.5%) cases of HCC, respectively. Immunohistochemical staining showed strong fine granular intracytoplasmic staining in tumor cells, whereas we observed occasional weak staining in normal liver tissues surrounding the tumors. We found that 14 out of 37 (37.8%) HCC expressed moderate to strong FACL4 immunostaining. Both normal adult and fetal liver tissues showed very weak to no detectable staining, whereas 3 out of 10 (30%) cirrhotic livers expressed weak staining. In addition, we found that 4 out of 8 (50%) human hepatoma cell lines expressed high levels of FACL4 by northern blot analysis. Our results show that FACL4 is a new molecular marker for HCC and suggest that the FACL4 pathway may be involved in liver carcinogenesis. (Cancer Sci 2003; 94: 421–424)

Cloning of BNIP3h, a member of proapoptotic BNIP3 family genes.

Apoptosis is regulated by interaction of antiapoptotic Bcl-2 family proteins with various proapoptotic proteins, several of which are also members of the Bcl-2 family. BNIP3 (formerly NIP3) is a proapoptotic mitochondrial protein classified in the Bcl-2 family based on limited sequence homology-3 (BH3) domain and COOH-terminal transmembrane domain. Sequence comparison of BNIP3 has indicated that there are several BNIP3 human homologs of this protein, like BNIP3L, Nix and BNIP3. We have cloned a new member of BNIP3 family from the cDNA library prepared from human dermal papilla cells and designated as BNIP3h. BNIP3h shows substantial homology with other BNIP3 family proteins. BNIP3h induced apoptosis from 24 hours after transfection in MCF7 cell lines and its apoptosis inducing activity is extended until 72 hours after transfection.

Risk factors associated with metabolic syndrome in Korean elderly.

Objective: The aim of our study was to identify risk factors related to the prevalence of metabolic syndrome (MetS) in the Korean elderly population. Methods: This cross-sectional study was conducted with 404 Korean elderly subjects (118 male, 286 female), aged 60 years or older between 2000 and 2003. The subjects were divided into MetS and control groups based on Adult Treatment Panel III guidelines and by applying the Asia-Pacific waist circumference. Dietary intake, blood profiles, and anthropometric variables were measured. Results: The MetS group had a significantly higher serum triglyceride level (p < 0.001) than the control group. The risk of the MetS was inversely associated with the intake of protein, fat, vitamin A, and vitamin E in women. Plasma levels of interleukin-1RA were higher in the MetS group (p < 0.05); however, interleukin-2 levels were not significantly different. Plasma antioxidant vitamin levels tended to be lower in the MetS group and were negatively associated with the risk factors of MetS. Conclusion: Korean elderly subjects with MetS tended to have an increased inflammatory response and a decreased antioxidant capacity. Furthermore, it was identified that undernutrition (especially a deficiency of antioxidant vitamins), not excess intake of energy, fat, or cholesterol, was associated with risk for MetS.

Expression pattern and intensity of protoporphyrin IX induced by liposomal 5-aminolevulinic acid in rat pilosebaceous unit throughout hair cycle.

We have developed liposomal formulation of 5-aminolevulinic acid (ALA) to enhance topical delivery and examined ALA-induced protoporpyrin (PpIX) expression in rat pilosebaceous unit throughout hair cycle. Two types of liposomes—glycerol dilaulate (GDL) and phosphatidylcholine (PC)—were formulated and both liposomal ALA increased PpIX expression in rat dorsal skin and pilosebaceous units when compared with free ALA. However, iontophoresis combined with liposomal ALA reduced the expression intensity of PpIX in hair bulbs although it achieved deeper and wider expression of PpIX through transfollicular pathway. After topical application in intact or depilated rat skin, liposomal ALA produced excellent PpIX expression in pilosebaceous units. The expression pattern and intensity of PpIX changed in hair cycle-dependent manner: specific expression only in sebaceous glands was observed at telogen; strong expression in whole pilosebaceous units was shown at anagen with intense expressions in hair bulbs and sebaceous glands; and a pattern similar to anagen but reduced intensity in the hair bulbs was seen at catagen. Throughout hair cycle, the expression pattern and intensity were dramatically changed in hair follicular epithelial cells depending on the cell density and proliferation activity of those cells, whereas those were consistent in sebaceous glands regardless of hair cycle. Little expression was shown in dermis. Photoactivation effect of 20% liposomal ALA-PDT using a red filtered-halogen lamp damaged sebaceous glands, hair follicles and epidermal layers. Formation of a thicker epidermal layer was observed, and hair induction after depilation was inhibited along with damage in sebaceous glands.

Detection of human papillomavirus 60 in epidermal cysts of nonpalmoplantar location

Human papillomavirus (HPV) types 57 and 60 are associated with epidermal cysts of palmoplantar location (PPECs). Recently, there was a report of HPV 60 in a wart located in a nonpalmoplantar area as a possible precursor of a nonpalmoplantar epidermal cyst (NPPEC). There has been no study that has examined the presence and frequency of HPV in ordinary NPPECs. We reviewed 63 cases of epidermal cysts, including 59 NPPECs and 4 PPECs. After routine histopathologic review, polymerase chain reaction (PCR) of extracted DNA, sequencing of PCR products, and, finally, a homology search were undertaken. As a result, 9 of 63 (14.3%) cysts, including 6 of 59 NPPECs (10.2%), showed a positive reaction on PCR. By homology search after PCR, all were demonstrated as HPV 60. For NPPECs, no variables, including some histopathologic features, are significantly related to the presence of HPV 60 in univariate or multivariate analysis (P > 0.05).

Effect of heat shock on the vascular contractility in isolated rat aorta.

Stresses such as heat shock, ischemia, and irradiation have been known to induce heat shock proteins in various tissues. We investigated the effects of heat shock on the vascular contractility by using isolated rat aorta. Rat thoracic aortic rings were mounted in an organ bath maintained at 37 degrees C. For heat shock, aortic ring preparations were exposed to 42 degrees C for either 15 or 45 min (heat shock group), whereas the control group was left at 37 degrees C. Eight hours after heat shock, aortic ring preparations were subjected to contractions with high K(+) membrane-depolarizing solution. After functional study, tissues were frozen for measurement of heat shock protein 70 (HSP70). Heat shock not only increased the expression of HSP70 in the rat aorta, but also augmented contractions to KCl whether endothelium was present or denuded. Short exposure of tissues to 42 degrees C for 15 min did not work either. These results suggest that heat shock increases vascular contractility in isolated aortic strips.

Comparative analysis of bacterial diversity and communities inhabiting the fairy ring of Tricholoma matsutake by barcoded pyrosequencing

Comparative analysis of the soil bacterial communities inhabiting the fairy ring of Tricholoma matsutake.