Carnosol induces apoptotic cell death through ROS-dependent inactivation of STAT3 in human melanoma G361 cells

Melanoma is the leading cause of skin cancer deaths, and the poor prognosis of metastatic melanoma has made needs for a novel pharmacological treatment or efficient intervention. Carnosol, a major polyphenolic compound from Rosmarinus officinalis, has a wide range of biological activities including anti-cancer effect. However, the underlying molecular mechanisms of its anti-cancer effect remain poorly understood in malignant human melanoma cells. In the present study, we investigate the apoptotic effect and the underlying anti-cancer mechanisms of carnosol. Our results revealed that carnosol strongly induced apoptosis against human melanoma G361 cells in a dose- and time-dependent manner, and caused dramatical elevation in cellular reactive oxygen species (ROS) level during apoptosis. In mechanistic studies, carnosol treatment decreased protein level of anti-apoptotic B‑cell lymphoma 2 (Bcl-2) and B cell lymphoma-extra large (Bcl-xL), however, increased level of pro-apoptotic Bcl-2-associated X protein (Bax) protein. Moreover, carnosol escalated cellular level of p53, which was accompanied by a decline of mouse double minute 2 homolog (MDM2) level. Also, carnosol inhibited activation of Src and signal transducer and activator of transcription 3 (STAT3), therefore down-regulated STAT3-dependent gene expression, such as D-series cyclin and survivin. These changes by carnosol were attenuated by pre-treatment of N-acetyl cysteine, and abolished progression of carnosol-induced apoptosis. In conclusion, carnosol induced apoptosis in human melanoma G361 cells through ROS generation and inhibition of STAT3-mediated pathway. Our results provide molecular bases of carnosol-induced apoptosis, and suggest a novel candidate for human melanoma treatment.This research was supported by Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Education (2018R1D1A1A02050495, J.-S. Choi) and by the Ministry of Science, ICT and Future Planning 2017R1A2B4009831, K.- S. Chun)

Synergistic Effect between Cryptotanshinone and Antibiotics against Clinic Methicillin and Vancomycin-Resistant Staphylococcus aureus

Cryptotanshinone (CT), a major tanshinone of medicinal plant Salvia miltiorrhiza Bunge, demonstrated strong antibacterial activity against clinic isolated methicillin and vancomycin-resistant Staphylococcus aureus (MRSA and VRSA) in this experiment. The CT was determined against clinic isolated MRSA 1–16 with MIC and MBC values ranging from 4 to 32 and 8 to 128 μg/mL; for MSSA 1-2 from 16 to 32 μg/mL and 64 to 128 μg/mL; for VRSA 1-2 from 2 to 4 μg/mL and 4 to 16 μg/mL, respectively. The range of MIC50 and MIC90 of CT was 0.5–8 μg/mL and 4–64 μg/mL, respectively. The combination effects of CT with antibiotics were synergistic (FIC index <0.5) against most of tested clinic isolated MRSA, MSSA, and VRSA except additive, MRSA 4 and 16 in oxacillin, MRSA 6, 12, and 15 in ampicillin, and MRSA 6, 11, and 15 in vancomycin (FIC index < 0.75–1.0). Furthermore, a time-kill study showed that the growth of the tested bacteria was completely attenuated after 2–6 h of treatment with the 1/2 MIC of CT, regardless of whether it was administered alone or with ampicillin, oxacillin, or vancomycin. The results suggest that CT could be employed as a natural antibacterial agent against multidrug-resistant pathogens infection

Effects of dodecacalcium heptaaluminate content on the setting time, compressive strength, alkalinity, and cytocompatibility of tricalcium silicate cement

Objective: This study aimed to investigate the effects of dodecacalcium hepta-aluminate (C12A7) content on some physicochemical properties and cytocompatibility of tricalcium silicate (C3S) cement using human dental pulp cells (hDPCs). Material and Methods: High purity C3S cement was manufactured by a solid phase method. C12A7 was mixed with the cement in proportions of 0, 5, 8, and 10 wt% (C12A7-0, -5, -8, and -10, respectively). Physicochemical properties including initial setting time, compressive strength, and alkalinity were evaluated. Cytocompatibility was assessed with cell viability tests and cell number counts. Statistical analysis was performed by using one-way analysis of variance (ANOVA) and Tukey’s test (p&lt;0.05). Results: The initial setting time of C3S-based cement was shorter in the presence of C12A7 (p&lt;0.05). After 1 day, C12A7-5 showed significantly higher compressive strength than the other groups (p&lt;0.05). After 7 days, the compressive strength of C12A7-5 was similar to that of C12A7-0, whereas other groups showed strength lower than C12A7-0. The pH values of all tested groups showed no significant differences after 1 day (p&gt;0.05). The C12A7-5 group showed similar cell viability to the C12A7-0 group (p&gt;0.05), while the other experimental groups showed lower values compared to C12A7-0 group (p&lt;0.05). The number of cells grown on the C12A7-5 specimen was higher than that on C12A7-8 and -10 (p&lt;0.05). Conclusions: The addition of C12A7 to C3S cement at a proportion of 5% resulted in rapid initial setting time and higher compressive strength with no adverse effects on cytocompatibility

Development of Single-Walled Carbon Nanotube-Based Biosensor for the Detection of Staphylococcus aureus

The goal of this research is to develop a single-walled carbon nanotube- (SWCNT-) based biosensor to detect Staphylococcus aureus. The specificity of 11 bacteria and polyclonal anti-Staphylococcus aureus antibodies (pAbs) was determined using an indirect ELISA. The pAbs were immobilized onto sensor platform after the hybridization of 1-pyrenebutanoic acid succinimidyl ester (PBASE). The resistance difference (ΔR) was calculated using a potentiostat. The bacteria detected by the biosensor were observed using a scanning electron microscope (SEM). The optimum concentration of SWCNTs on the platform was determined to be 0.1 mg/mL. The binding of pAbs with S. aureus resulted in a significant increase in resistance value of the biosensor (P<0.05). The SEM images confirmed the specific binding of S. aureus on the biosensor. The SWCNT-based biosensor was able to detect S. aureus with a limit of detection (LOD) of 4 log⁡CFU/mL

A Study on Apparel Design for Chiropractic Training

In severe cases of spinal misalignment due to wrong back posture during adolescence or focusing on work, modern people can have back pain. Such a symptom is commonly referred to as scoliosis and it already became one of the lifestyle diseases. Hence, the purpose of the present study was to develop a functional musculoskeletal correction wear that can help with maintaining a correct posture by relaxing tonic muscles and maximizing the tension of the relaxed muscle using the principle of sports taping. Market research for design development was conducted at department stores and specialty road shops separately. Department stores were visited on weekdays when the store is not crowded, and specialty road shops were visited at around three o’clock on weekdays after checking their opening, lunch, and closing times. Since the apparel should not interfere with exercise movements while maintaining the body shape, the survey was carried out mostly on yoga and lifestyle training apparel focusing on elasticity and percentage of blending fiber. A camera and audio recorder were prepared and took pictures of clothing tags that show the ratio of polyester and spandex while trying on and checking products at the stores. Data were collected by asking the staff at road specialty shops who has more professional knowledge about the advantages and functions of the existing products and recording their answers. Point color schemes in black color family in which body shapes can look beautiful were mostly checked. Due to the design characteristics that have to maintain the spine, the designs of all-in-one products that can maintain the upper and lower torsos were checked at underwear stores. The existing products were evaluated for their designs to be adopted including the material, integrated into the new design, and then modified and supplemented, and the following chiropractic apparel design was drawn (Table 1). An all-in-one design that wraps around the hips and upper thighs for easy counterbalancing between the upper and lower body was chosen. A design with an accent line that makes body shape looks beautiful and Used compression wear material that is comfortable for activities and presses tight against the skin. Activates the muscle activities of latissimus dorsi muscle using taping method, and provides tension to maintain correct posture of round back and shoulders. In conclusion, prevention, exercise, and stretching are important for scoliosis and sound posture for the loin should be constantly maintained. To that end, not only the proper use of chairs but also apparel that is worn all the time can be helpful

Uterine and placental expression of TRPV6 gene is regulated via progesterone receptor- or estrogen receptor-mediated pathways during pregnancy in rodents

Transient receptor potential cation channel, subfamily V, member 6 (TRPV6) is an epithelial Ca2+ channel protein expressed in calcium absorbing organs. In the present study, we investigated the expression and regulation of uterine and placental TRPV6 during gestation in rodents. Uterine TRPV6 peaked at pregnancy day (P) 0.5, P5.5 and, P13.5 and was detected in uterine epithelium and glands of rats, while placental TRPV6 mRNA levels increased in mid-gestation. Uterine and placental TRPV6 mRNA levels in rats appear to cyclically change during pregnancy, suggesting that TRPV6 may participate in the implantation process. In addition, uterine TRPV6 mRNA is only expressed in placenta-unattached areas of the uterus, and uterine TRPV6 immunoreactivity was observed in luminal and glandular epithelial cells. In the placenta, TRPV6 was detected in the labyrinth and spongy zone. These results may indicate that TRPV6 has at least two functions: implantation of the embryo and maintenance of pregnancy. To investigate the pathway(s) mediating TRPV6 expression in rodents, anti-steroid hormone antagonists were injected prior to maximal TRPV6 expression. In rats, TRPV6 expression was reduced by RU486 (an anti-progesterone) through progesterone receptors, and ICI 182,780 (an anti-estrogen) blocked TRPV6 expression via estrogen receptors in mice. The juxtaposition of uterine and placental TRPV6 expressed in these tissues supports the notion that TRPV6 participates in transferring calcium ions between the maternal and fetal compartments. Taken together, TRPV6 gene may function as a key element in controlling calcium transport in the uterus between the embryo and the placenta during pregnancy

Increasing late diagnosis in HIV infection in South Korea: 2000-2007

<p>Abstract</p> <p>Background</p> <p>The number of Koreans diagnosed with human immunodeficiency virus (HIV) infections is increasing annually; however, CD4+ T-cell counts at diagnosis have decreased. The purpose of the present study was to identify clinical and epidemiologic associations with low CD4+ T-cell counts at the time of HIV diagnosis in a Korean population.</p> <p>Methods</p> <p>Data from 2,299 HIV-infected individuals with initial CD4+ T-cell counts measured within 6 months of HIV diagnosis and reason for HIV testing were recorded and measured from 2000 to 2007. Data were selected from the database of the Korea Centers for Disease Control and Prevention. Late diagnosis was defined by CD4+ T-cell counts <200 cells/mm³. Reasons for HIV testing were analyzed using logistic regression including epidemiologic variables.</p> <p>Results</p> <p>A total of 858 individuals (37.3%) were included in the late diagnosis group. Individuals with a late diagnosis were older, exposed through heterosexual contact, and demonstrated clinical manifestations of acquired immunodeficiency syndrome (AIDS). The primary reason for HIV testing was a routine health check-up (41%) followed by clinical manifestations (31%) of AIDS. The proportion of individuals with a late diagnosis was higher in individuals tested due to clinical symptoms in public health centers (adjusted odds ratio [AOR], 17.3; 95% CI, 1.7-175) and hospitals (AOR, 4.9; 95% CI, 3.4-7.2) compared to general health check-up. Late diagnosis annually increased in individuals diagnosed by voluntary testing both in public health centers (PHCs, P = 0.017) and in hospitals (P = 0.063). Routine testing due to risky behaviors resulted in earlier detection than testing secondary to health check-ups, although this difference was not statistically significant (AOR, 0.7; P = 0.187). Individuals identified as part of hospital health check-ups more frequently had a late diagnosis (P = 0.001)</p> <p>Conclusions</p> <p>HIV infection was primarily detected by voluntary testing with identification in PHCs and by testing due to clinical symptoms in hospitals. However, early detection was not influenced by either voluntary testing or general health check-up. It is important to encourage voluntary testing for early detection to decrease the prevalence of HIV infection and AIDS progression.</p

Use of signal sequences as an in situ removable sequence element to stimulate protein synthesis in cell-free extracts

This study developed a method to boost the expression of recombinant proteins in a cell-free protein synthesis system without leaving additional amino acid residues. It was found that the nucleotide sequences of the signal peptides serve as an efficient downstream box to stimulate protein synthesis when they were fused upstream of the target genes. The extent of stimulation was critically affected by the identity of the second codons of the signal sequences. Moreover, the yield of the synthesized protein was enhanced by as much as 10 times in the presence of an optimal second codon. The signal peptides were in situ cleaved and the target proteins were produced in their native sizes by carrying out the cell-free synthesis reactions in the presence of Triton X-100, most likely through the activation of signal peptidase in the S30 extract. The amplification of the template DNA and the addition of the signal sequences were accomplished by PCR. Hence, elevated levels of recombinant proteins were generated within several hours

COMPARISON OF THE RISK FACTORS OF KOREAN ADOLESCENT SUICIDE RESIDING IN HIGH SUICIDAL REGIONS VERSUS THOSE IN LOW SUICIDAL REGIONS

Background: The suicide rate of the youth in South Korea has been increasing, and suicide of the youth still has been the most common cause of death since 2007. We aimed to determine the trends and the regional risk factors of youth suicide in South Korea from 2001 to 2010. Subjects and Methods: We used the data from the National Statistical Office to calculate the standardized suicide rates and various regional data including population census, employment, and labor. To calculate the effect of individual risk factors, we used the data from the fourth Korean Youth Risk Behavior Web-based Survey (KYRBWS-VI). Conditional autoregressive model for regional standardized mortality ratio (SMR) using inter-regional spatial information was fitted. Results: Suicide rates of adolescents aged 12 to 18 was from 3.5 per 100,000 people in 2001 and 5.3 per 100,000 in 2010. There were no significant gender difference in suicide rates, however, the number of suicides among adolescents aged 15-18 accounted for four times than those of adolescents ages 12-14. High proportion of late adolescents, higher number of recipients of national basic livelihood, and higher number of adolescents who treated with depression were related to elevated suicide rate of adolescent. Total sleep time of adolescents and regional unemployment rate were negatively associated with the suicide risk of respective regions. Conclusions: Age distribution, economic status, total sleep time, and the number of adolescent patients with depression were different between those in low and in high adolescent suicidal regions in Korea. Our findings suggest that preferential appliance of adolescent suicide prevention program for regions by considering those factors may be important steps to reduce adolescent suicide in Korea