This study describes the characterization of 80 kDa protease showing gelationlytic property among three proteases in the excretory/secretory proteins (ESP) from Toxoplasma gondii. The protease activity was detected in the ESP but not in the somatic extract of RH tachyzoites. This protease was active only in the presence of calcium ion but not other divalent cationic ions such as Cu (2+), Zn (2+), Mg (2+), and Mn (2+), implying that Ca (2+) is critical factor for the activation of the protease. The 80 kDa protease was optimally active at pH 7.5. Its gelatinolytic activity was maximal at 37 degrees C, and significant level of enzyme activity of the protease remained after heat treatment at 56 degrees C for 30 min or 100 degrees C for 10 min. This thermostable enzyme was strongly inhibited by metal chelators, i.e., EDTA, EGTA, and 1, 10-phenanthroline. Thus, the 80 kDa protease in the ESP secreted by T. gondii was classified as a calcium dependent neutral metalloprotease.
Animals ; Calcium/metabolism ; Endopeptidases/*metabolism ; Hydrogen-Ion Concentration ; Molecular Weight ; Temperature ; Toxoplasma/*enzymology

Authors review the clinical and histopathologic findings of 41 EIenoch-Schonlein purpur.a patients and attempt, to clarify the association between abnorma] urinary findings and anti-streptolysin 0(ASLO) titer and performed the direct immunofluorescence(',E)II) study on skin lesion to investigate the pathogenetic mechanim of renal involvement. The results were summarized as follows : 1. Ag(, distribution was form 3 month to 63 years and mean age was 18.9 years. male to female ratio was 1.2:1(22 males '. 19 females). 2. In he past history, there were preceding upper respora.tory infections in 25(61:o)patients and drug history in 10(24.3%)patients. 3. (linical manifestations in the order of frequency were as follows : arthralgia (60.9%), hematuria or albuminuria(43.8%), abdominal pain(21.9%), and positive occult: blood test(7.3%)in stool examination. 4. In the laboratory fiindings, leukocytosis was cleveloped in 14.6%, anemia in 2.4%, increased eryttrocyte sec1imentation zate in 12.3% increased ASLG titer in 43.9% of patients, and serum IgG was increased in 4 of 20(20.6)patierits, IgA in 4 of 21(19%)patients, IgM in 1fo 19(5.3o)patients. 5. Because correlation between increased A.LO titer ancl ahnoormal urinary findings wa.s high(x-test, p< 0.0,5), increased ASI,O titer may be inclicator of renal involvement. 6. DIF stuc].y was per formed in 34 patients. All showed positive reactiviy. IgG(17%), IgA(29.4% ), Clq(14.6%), C3(12.2% ), fibrinog.n(82.4%) were deposited around the vessel wall in a granular pattern. 7. There was statistically significant increase in mean serum IgG value in patients with abnormal urinary findings( t test, p < 0.05 ).
Anemia ; Arthralgia ; Female ; Hematuria ; Humans ; Immunoglobulin A ; Immunoglobulin G ; Immunoglobulin M ; Leukocytosis ; Male ; Purpura* ; Purpura, Schoenlein-Henoch ; Skin

BACKGROUND/AIMS: The purpose of this study was to evaluate the effectiveness of lamivudine treatment in patients with chronic liver disease caused by chronic infection of hepatitis B virus (HBV). METHODS: Thirty-ive patients with chronic infection of HBV were included in this study who were diagnosed at Hanyang University Hospital from January 1998 to January 1999. They received 150mg of lamivudine per oral once daily for 6 months with follow-p of liver function test, serum HBV DNA and serologic markers for hepatitis B virus every two months. Lamivudine was well tolerated. Eight patients underwent liver biopsies before entering the study and follow-p biopsies were done at 5 patients. RESULTS: Out of all 35 patients, chronic hepatitis patients histologically confirmed were 8, chronic hepatitis patients clinically diagnosed were 25 and liver cirrhosis patients clinically diagnosed were 2. The mean age was 35.7 years. Male-female ratio was 2.2:1. There was no hepatitis B surface antigen (HBsAg) negative seroconversion. The HBeAg loss rate was 26.9%(7/26) and HBeAg seroconversion rate was 10.7%(3/28) at the end of follow-p. Ten patients were anti-Be positive prior to treatment, 3 of them became anti-Be negative at the end of follow-p. Five patients underwent follow-p liver biopsies, in which histologic improvements were shown in 4 cases. Serum replicative HBV DNA by bDNA assay was decreased in all patients and HBV DNA was undetectable in 52.9%(9/17) at the end of treatment. Out of the 15 patients with abnormal alanine aminotransferase (ALT) levels at baseline, ALT level in 7 patients(46.7%) was normalized at treatment completion. Pretherapy ALT level was the only predictive factor for loss of HBeAg by stepwise logistic regression analysis(odds ratio : 1.0208) (95% Confidence Interval : 1.0023 ~ 1.0396) (p value=0.0271). CONCLUSIONS: Lamivudine induced sustained suppression of HBV replication during treatment in all patients. In treating patients with lamivudine, who had chronic liver disease due to chronic infection of HBV, the improvement of liver function test and suppression of viral replication appeared early and was sustained during the 6months treatment. This, in turn, may induce histological improvement as well. Pretherapy ALT level was the only predictive determinant for HBeAg loss during lamivudine therapy, and that should be kept in mind in selecting patients for treatment.
Alanine Transaminase ; Biopsy ; Branched DNA Signal Amplification Assay ; DNA ; Hepatitis B e Antigens ; Hepatitis B Surface Antigens ; Hepatitis B virus ; Hepatitis B* ; Hepatitis* ; Hepatitis, Chronic ; Humans ; Lamivudine* ; Liver ; Liver Cirrhosis ; Liver Diseases ; Liver Function Tests ; Logistic Models ; Prospective Studies*

Skin photoaging occurs due to chronic exposure to solar ultraviolet radiation (UV), the main factor contributing to extrinsic skin aging. Clinical signs of photoaging include the formation of deep, coarse skin wrinkles and hyperpigmentation.Although melanogenesis and skin wrinkling occur in different skin cells and have different underlying mechanisms, their initiation involves intracellular calcium signaling via calcium ion channels. The ORAI1 channel initiates melanogenesis in melanocytes, and the TRPV1 channel initiates MMP-1 production in keratinocytes in response to UV stimulation. We aimed to develop a drug that may simultaneously inhibit ORAI1 and TRPV1 activity to help prevent photoaging. We synthesized nootkatol, a chemical derivative of valencene. TRPV1 and ORAI1 activities were measured using the whole-cell patch-clamp technique. Intracellular calcium concentration [Ca2+ ] i was measured using calcium-sensitive fluorescent dye (Fura-2 AM). UV-induced melanin formation and MMP-1 production were quantified in B16F10 melanoma cells and HaCaT cells, respectively. Our results indicate that nootkatol (90 μM) reduced TRPV1 current by 94% ± 2% at –60 mV and ORAI1 current by 97% ± 1% at –120 mV. Intracellular calcium signaling was significantly inhibited by nootkatol in response to ORAI1 activation in human primary melanocytes (51.6% ± 0.98% at 100 μM). Additionally, UV-induced melanin synthesis was reduced by 76.38% ± 5.90% in B16F10 melanoma cells, and UV-induced MMP-1 production was reduced by 59.33% ± 1.49% in HaCaT cells. In conclusion, nootkatol inhibits both TRPV1 and ORAI1 to prevent photoaging, and targeting ion channels may be a promising strategy for preventing photoaging.

Skin photoaging occurs due to chronic exposure to solar ultraviolet radiation (UV), the main factor contributing to extrinsic skin aging. Clinical signs of photoaging include the formation of deep, coarse skin wrinkles and hyperpigmentation.Although melanogenesis and skin wrinkling occur in different skin cells and have different underlying mechanisms, their initiation involves intracellular calcium signaling via calcium ion channels. The ORAI1 channel initiates melanogenesis in melanocytes, and the TRPV1 channel initiates MMP-1 production in keratinocytes in response to UV stimulation. We aimed to develop a drug that may simultaneously inhibit ORAI1 and TRPV1 activity to help prevent photoaging. We synthesized nootkatol, a chemical derivative of valencene. TRPV1 and ORAI1 activities were measured using the whole-cell patch-clamp technique. Intracellular calcium concentration [Ca2+ ] i was measured using calcium-sensitive fluorescent dye (Fura-2 AM). UV-induced melanin formation and MMP-1 production were quantified in B16F10 melanoma cells and HaCaT cells, respectively. Our results indicate that nootkatol (90 μM) reduced TRPV1 current by 94% ± 2% at –60 mV and ORAI1 current by 97% ± 1% at –120 mV. Intracellular calcium signaling was significantly inhibited by nootkatol in response to ORAI1 activation in human primary melanocytes (51.6% ± 0.98% at 100 μM). Additionally, UV-induced melanin synthesis was reduced by 76.38% ± 5.90% in B16F10 melanoma cells, and UV-induced MMP-1 production was reduced by 59.33% ± 1.49% in HaCaT cells. In conclusion, nootkatol inhibits both TRPV1 and ORAI1 to prevent photoaging, and targeting ion channels may be a promising strategy for preventing photoaging.

Juvenile xanthogranuloma(JXG) is a a benign histiocytic proliferative disorder most frequently seen in infants. The usual course is self-limited, but it may be a manifestation of juvenile chronic myeloid leukemia(JCML). In addition to JXG, some patient with JCML may have multiple cafe-au-lait spots and a family history of neurofibromatosis(NF). The association of JXG and cafe-au-lait spots in patients with JCML is of interest in light of description of otherwise healthy children with JXG who also had cafe-au-lait spots. We report herein a case of JXG in 18-month-old boy who also had multiple cafe-au-alit spots, which predated the JCML.
Cafe-au-Lait Spots ; Child ; Humans ; Infant ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive* ; Male ; Neurofibromatoses* ; Xanthogranuloma, Juvenile*

The dysplasitc melanocytic nevus(DMN) of the skin is an atypical-appearing melanocytic tumor characterized by intraepidermal atypical melanocytic preliferation. Clinically, DMN has a diameter greater than 5mm, ill-defined irregular borders, irregularly distributed pigmentation surrounded by erythems,. Histopat- hologically, there are numerous discrete nests of atypical melanocytes situated at the dermoepidermal junction of elongated rete ridges. In the dermis, concentric lamellar fibroplasia, lymphocytic infiltrates and neovascularization are noted. We report herein a case of DMN developed on the trunk of 31-year-old Korean male.
Adult ; Dermis ; Humans ; Male ; Melanocytes ; Nevus, Pigmented* ; Pigmentation ; Skin

BACKGROUND: This study was designed to examine the interactions between mivacurium and vecuronium when administered during a standardized technique. METHODS: Eighty patients (ASA physical status I or II) were randomly assigned to one of four groups (n=20). Their neuromuscular function was measured in response to ulnar nerve supramaximal square-wave TOF stimulation at 10-sec intervals. After the attainment of a stable baseline neuromuscular response, the patients were randomly assigned to receive a rapid iv bolus of either: (1) 3M group (n=20): mivacurium 0.21 mg/kg. Alone, or (2) 2M1V group (n=20): mivacurium 0.14 mg/kg plus vecuronium 0.05 mg/kg, or (3) 1M2V group (n=20): mivacurium 0.07 mg/kg plus vecuronium 0.10 mg/kg, or (4) 3V group (n=20): vecuronium 0.15 mg/kg alone. The onset time of the neuromuscular block, time of recovery of T1 to 25% and reblock time (the time from the reinjection of vecuronium at the time of recovery of T1 to 25% to the time of recovery of T1 to 25%: T25-25) were measured. The intubating condition was evaluated clinically with a scoring system. RESULTS: The onset of block in the 3M group was 33% slower than in the 3V group. The time durations until 25% recovery in the 2M1V, 1M2V and 3V groups were longer than in the 3M group, and the durations in the 1M2V and 3V groups were longer than in the 2M1V group. The T25-25 reblock times of the 2M1V, 1M2V and 3V groups were prolonged in comparison to that of the 3M group. There was no difference in intubating conditions between any of the groups. CONCLUSIONS: A combination of mivacurium with vecuronium provides rates of onset and duration of block which are more effective than an equivalent dose of mivacuriun alone as an additive reaction.
Anesthesia, General* ; Humans ; Neuromuscular Blockade* ; Ulnar Nerve ; Vecuronium Bromide*

C-erbB-2, one of epidermal growth factor receptor gene family, may have an important role in progression of transitional cell carcinoma (TCC of the urinary bladder. We herein immunohistochemically examined 52 bladder TCC specimens for expression of c-erbB-2 gene product to investigate its prognostic value. Interrelationship between expression of c-erbB-2 and stage, grade, expression of proliferating cell nuclear antigen (PCNA), and clinical outcomes were analyzed. Overexpression of c-erbB-2 was significantly higher in invasive tumors as compared with superficial tumors and in high grade tumors as compared with low grade tumors (p<0.005). However, there were no significant correlations between the degree of expression of c-erbB-2 and progression-free survival in patients with superficial bladder TCC as well as actual-survival in patients with invasive bladder TCC. There also was no significant correlation between the expression of c-erbB-2 and PCNA. These results suggest that c-trbB-2 may be an important marker of malignant potentials and invasiveness of bladder TCC, but immunohistochemical study for its expression in bladder TCC may not provide additional prognostic information to stage and grade of the tumors.
Carcinoma, Transitional Cell ; Disease-Free Survival ; Genes, erbB-2 ; Humans ; Proliferating Cell Nuclear Antigen ; Receptor, Epidermal Growth Factor ; Urinary Bladder Neoplasms* ; Urinary Bladder*

Objective: To examine the effects of Tribulus terrestris L. (T. terrestris) extract on the modulation of calcium channels to evaluate its use in topical agents for treatment of atopic dermatitis. Methods: The 70% methanol extract of T. terrestris was prepared. Human HEK293T cells with over-expressed calcium release-activated calcium channel protein 1 (Orai1), transient receptor potential vanilloid 1, or transient receptor potential vanilloid 3 (TRPV3) were treated with T. terrestris extract. Modulation of ion channels was measured using a conventional whole-cell patch-clamp technique. Results: T. terrestris extract (100 mg/mL) significantly inhibited Orai1 activity in Orai1-stromal interaction molecule 1 co-overexpressed HEK293T cells. In addition, T. terrestris extract significantly increased the TRPV3 activity compared with 2-Aminoethyl diphe-nylborinate (100 mmol/L), which induces the full activation of TRPV3. Conclusions: Our results suggest that T. terrestris extract may have a therapeutic po-tential for recovery of abnormal skin barrier pathologies in atopic dermatitis through modulating the activities of calcium ion channels, Orai1 and TRPV3. This is the first study to report the modulatory effect of a medicinal plant on the function of ion channels in skin barrier.