Calf Circumference as an Indicator for Cystatin C Testing in Hospitalized Elderly Male Patients for Detecting Hidden Renal Impairment.

Serum creatinine is used to measure the estimated glomerular filtration rate (eGFR); however, it is influenced by muscle mass and may therefore overestimate renal function in patients with sarcopenia. We examined calf circumference (CC) as a convenient muscle mass evaluation tool that can potentially indicate the need to test for cystatin C instead of creatinine in elderly inpatients. We retrospectively reviewed the electronic health record of 271 inpatients aged 65 or over. CC was determined by measuring the thickest part of the nondominant calf. eGFRcys and eGFRcr were calculated using cystatin C and creatinine levels, respectively. We evaluated optimum CC cutoff values using the eGFRcys/eGFRcr ratio for detecting hidden renal impairment (HRI, defined as eGFRcr ≥ 60 mL/min/1.73 m2 but eGFRcys < 60 mL/min/1.73 m2). CC showed a significant positive correlation with the eGFRcys/eGFRcr ratio in both sexes. The areas under the receiver operating characteristic curve were 0.725 and 0.681 for males and females, respectively. CC cutoffs with a sensitivity or specificity of 90% or 95% might be used to detect HRI in males. In conclusion, utilizing the optimum cutoff, CC could be a cost-effective screening tool for detecting HRI in elderly male patients using cystatin C as an add-on test.

Decoding trajectories of imagined hand movement using electrocorticograms for brain-machine interface.

Objective. Reaching hand movement is an important motor skill actively examined in the brain-computer interface (BCI). Among the various components of movement analyzed is the hand's trajectory, which describes the hand's continuous positions in three-dimensional space. While a large body of studies have investigated the decoding of real movements and the reconstruction of real hand movement trajectories from neural signals, fewer studies have attempted to decode the trajectory of the imagined hand movement. To develop BCI systems for patients with hand motor dysfunctions, the systems essentially have to achieve movement-free control of external devices, which is only possible through successful decoding of purely imagined hand movement.Approach. To achieve this goal, this study used a machine learning technique (i.e. the variational Bayesian least square) to analyze the electrocorticogram (ECoG) of 18 epilepsy patients obtained from when they performed movement execution (ME) and kinesthetic movement imagination (KMI) of the reach-and-grasp hand action.Main results. The variational Bayesian decoding model was able to successfully predict the imagined trajectories of the hand movement significantly above the chance level. The Pearson's correlation coefficient between the imagined and predicted trajectories was 0.3393 and 0.4936 for the KMI (KMI trials only) and MEKMI paradigm (alternating trials of ME and KMI), respectively.Significance. This study demonstrated a high accuracy of prediction for the trajectories of imagined hand movement, and more importantly, a higher decoding accuracy of the imagined trajectories in the MEKMI paradigm compared to the KMI paradigm solely.

Profiles of Non-aureus Staphylococci in Retail Pork and Slaughterhouse Carcasses: Prevalence, Antimicrobial Resistance, and Genetic Determinant of Fusidic Acid Resistance.

As commensal colonizers in livestock, there has been little attention on staphylococci, especially non-aureus staphylococci (NAS), contaminating meat production chain. To assess prevalence of staphylococci in retail pork and slaughterhouse carcass samples in Korea, we collected 578 samples from Korean slaughterhouses (n=311) and retail markets (n=267) for isolation of staphylococci and determined antimicrobial resistance phenotypes in all the isolates. The presence of and prevalence of fusB-family genes (fusB, fusC, fusD, and fusF) and mutations in fusA genes were examined in fusidic acid resistant isolates. A total of 47 staphylococcal isolates of 4 different species (Staphylococcus aureus, n=4; S. hyicus, n=1; S. epidermidis, n=10; Mammaliicoccus sciuri, n=32) were isolated. Fusidic acid resistance were confirmed in 9/10 S. epidermidis and all of the 32 M. sciuri (previously S. sciuri) isolates. Acquired fusidic acid resistance genes were detected in all the resistant strains; fusB and fusC in S. epidermidis and fusB/C in M. sciuri. Multi-locus sequence type analysis revealed that ST63 (n=10, 31%) and ST30 (n=8, 25%) genotypes were most prevalent among fusidic acid resistant M. sciuri isolates. In conclusion, the high prevalence of fusB-family genes in S. epidermidis and M. sciuri strains isolated from pork indicated that NAS might act as a reservoir for fusidic acid resistance gene transmissions in pork production chains.

Characterization of kinesthetic motor imagery compared with visual motor imageries.

Motor imagery (MI) is the only way for disabled subjects to robustly use a robot arm with a brain-machine interface. There are two main types of MI. Kinesthetic motor imagery (KMI) is proprioceptive (OR somato-) sensory imagination and Visual motor imagery (VMI) represents a visualization of the corresponding movement incorporating the visual network. Because these imagery tactics may use different networks, we hypothesized that the connectivity measures could characterize the two imageries better than the local activity. Electroencephalography data were recorded. Subjects performed different conditions, including motor execution (ME), KMI, VMI, and visual observation (VO). We tried to classify the KMI and VMI by conventional power analysis and by the connectivity measures. The mean accuracies of the classification of the KMI and VMI were 98.5% and 99.29% by connectivity measures (alpha and beta, respectively), which were higher than those by the normalized power (p < 0.01, Wilcoxon paired rank test). Additionally, the connectivity patterns were correlated between the ME-KMI and between the VO-VMI. The degree centrality (DC) was significantly higher in the left-S1 at the alpha-band in the KMI than in the VMI. The MI could be well classified because the KMI recruits a similar network to the ME. These findings could contribute to MI training methods.

The Effect of Degree of Milling on the Nutraceutical Content in Ecofriendly and Conventional Rice (Oryza sativa L.).

We investigated the effects of the type of rice and degree of milling (DOM) on the nutraceutical content and antioxidant activity of rice (Oryza sativa L.). The fatty acid (FA), vitamin E homolog, and phenolic contents in organic (OR), pesticide-free (PFR), and conventional rice (CR) decreased significantly with an increase in the DOM of rice grains, particularly for a DOM of 7 and 9 (p < 0.05). The 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity also decreased with the DOM; particularly, this activity decreased significantly, by approximately 60%, in rice grains with a DOM between 7 and 11, as compared to that of brown rice (p < 0.05). α-Tocopherol (r = 0.854) and p-coumaric acid (r = 0.501) showed the strongest correlation with DPPH activity in each chemical group. Stepwise discriminant analysis enabled the correct original and cross-validated classification of 87.0% and 81.5% of rice types, respectively. Additionally, the original and cross-validated classification of rice DOM levels showed that, overall, 93.8% and 92.6% of rice samples were correctly classified. Our findings reveal variations in the nutraceutical levels and antioxidant activities in rice grains based on the rice type and DOM, which can help improve the nutritional evaluation of human-health-promoting rice grains.

Triglyceride glucose index is a useful marker for predicting subclinical coronary artery disease in the absence of traditional risk factors.

BACKGROUND: Atherosclerotic cardiovascular (CV) events commonly occur in individuals with a low CV risk burden. This study evaluated the ability of the triglyceride glucose (TyG) index to predict subclinical coronary artery disease (CAD) in asymptomatic subjects without traditional CV risk factors (CVRFs). METHODS: This retrospective, cross-sectional, and observational study evaluated the association of TyG index with CAD in 1250 (52.8 ± 6.5 years, 46.9% male) asymptomatic individuals without traditional CVRFs (defined as systolic/diastolic blood pressure ≥ 140/90 mmHg; fasting glucose ≥126 mg/dL; total cholesterol ≥240 mg/dL; low-density lipoprotein cholesterol ≥160 mg/dL; high-density lipoprotein cholesterol < 40 mg/dL; body mass index ≥25.0 kg/m2; current smoking; and previous medical history of hypertension, diabetes, or dyslipidemia). CAD was defined as the presence of any coronary plaque on coronary computed tomographic angiography. The participants were divided into three groups based on TyG index tertiles. RESULTS: The prevalence of CAD increased with elevating TyG index tertiles (group I: 14.8% vs. group II: 19.3% vs. group III: 27.6%; P < 0.001). Multivariate logistic regression models showed that TyG index was associated with an increased risk of CAD (odds ratio [OR] 1.473, 95% confidence interval [CI] 1.026-2.166); especially non-calcified (OR 1.581, 95% CI 1.002-2.493) and mixed plaques (OR 2.419, 95% CI 1.051-5.569) (all P < 0.05). The optimal TyG index cut-off for predicting CAD was 8.44 (sensitivity 47.9%; specificity 68.5%; area under the curve 0.600; P < 0.001). The predictive value of this cut-off improved after considering the non-modifiable factors of old age and male sex. CONCLUSIONS: TyG index is an independent marker for predicting subclinical CAD in individuals conventionally considered healthy.

New Chemical Constituents from the Bark of Dendropanax morbifera Leveille and their Evaluation of Antioxidant Activities.

Four new constituents, as cis-6-oxogeran-4-enyl-10-oxy-O-ß-arabinopyranosyl-4'-O-ß-arabinopyranosyl-2''-octadec-9''',12''',15'''-trienoate (1), geran-3(10)-enyl-1-oxy-O-ß-arabinopyranosyl-4'-O-ß-arabinopyranosyl-2''-octadec-9''',12''',15'''-trienoate (2), geranilan-8-oxy-O-α-d-xylopyranosyl-2'-n-octadec-9'',12'',15''-trienoate (3), 1-cyclohex-2', 5'-dienyl 1-cyclohexylethanol-O-ß-d-xylopyranoside (4), along with six known constituents, guaiacol-O-ß-d-arabinopyaranoside (5), n-tetradecanyl oleate (6), oleyl-O-ß-d-xyloside (7), n-octadec-9,12-dienoyl-O-ß-d-arabinopyranoside (8), linolenyl-O-ß-d-arabinofuranoside (9) andglyceryl-1,3-dipalmito-2-olein (10), were isolated and identified from the Dendropanax morbifera bark. The new structures were established by one-and two-dimensional NMR (and in combination with IR, FAB-MSand HR-ESI-FTMS. The comparative evaluation of antioxidant potential by phosphomolybdenum, DPPH, FRAP and the NO assay of four different compounds (1-4), we have found that the compounds 1 and 2 have power as a natural antioxidant, whereas the compound 3 and 4 exhibited mild activity in comparison to compounds 1 and 2.

Relationship between the optimal cut-off values of anthropometric indices for predicting metabolic syndrome and carotid intima-medial thickness in a Korean population.

Anthropometric measurements are simple and useful methods for predicting metabolic syndrome (MetS) because obesity is a predominant feature of MetS. Although carotid intima-medial thickness (IMT) is generally used to evaluate subclinical atherosclerosis, the relationship between the optimal cut-off anthropometric values for predicting MetS and carotid IMT has not been analyzed in a Korean population.Anthropometric measurements including waist circumference (WC), waist hip ratio (WHR), waist height ratio (WHtR), body mass index (BMI), and carotid IMT were assessed in 2560 Korean subjects without previous history of cardiovascular disease, cerebrovascular disease, neurological abnormalities, or malignancy who participated in baseline health examinations in a self-referral setting in the Seoul area between April 2010 and November 2012. MetS was defined using the National Cholesterol Education Program-Adult Treatment Panel III criteria.In both men and women, the levels of all anthropometric indices were significantly higher in subjects with MetS than in those without MetS. According to the receiver operating characteristic curve, the values of 80.8 cm for WC, 0.87 for WHR, 0.52 for WHtR, and 24.6 kg/m for BMI were the optimal cut-offs for predicting MetS in women. The values of 89.3 cm for WC, 0.90 for WHR, 0.52 for WHtR, and 25.1 kg/m for BMI were the optimal cut-offs for predicting MetS in men. After adjusting for confounding factors, the WC optimal cut-off values for predicting MetS were independently associated with carotid IMT in both women and men (women: ß = 0.016, P = .008; men: ß = 0.033, P = .009). The optimal BMI cut-off value was independently associated with carotid IMT in men only (ß = 0.027, P = .032).Among anthropometric indices including WC, WHR, WHtR, and BMI, the WC optimal cut-off values for MetS were independently associated with an increased carotid IMT in both women and men in a Korean population.

High HDL-C levels reduce the risk of obstructive coronary artery disease in asymptomatic diabetics who achieved optimal glycemic control.

The benefit of a high level of high-density lipoprotein cholesterol (HDL-C) against coronary atherosclerosis risk after achieving optimal glycemic control (OGC) in diabetics remains uncertain. We aimed to evaluate the association between HDL-C and obstructive coronary artery disease (CAD) according to OGC status in diabetics. We analyzed 1,114 asymptomatic diabetics who underwent coronary computed tomographic angiography in a health examination. OGC was defined as hemoglobin A1C <7.0%. Obstructive CAD was defined as the presence of plaques with ≥50% stenosis. Patients with a high HDL-C level (≥40 mg/dL and ≥50 mg/dL in males and females, respectively) showed a lower prevalence of obstructive CAD than those with a low HDL-C level in the OGC group (8.9% vs. 14.4%; p = 0.046), but not in the non-OGC group (22.3% vs. 23.2%, p = 0.850). Multiple logistic regression models showed that the risk for obstructive CAD was lower in patients with a high HDL-C level than in those with a low HDL-C level in the OGC group (odds ratio: 0.584, 95% confidence interval: 0.343-0.995; p = 0.048), but not in the non-OGC group. In conclusion, it may be necessary to maintain a high HDL-C level to reduce the risk of obstructive CAD in asymptomatic diabetics after OGC is achieved.

Potential geo-discriminative tools to trace the origins of the dried slices of shiitake (Lentinula edodes) using stable isotope ratios and OPLS-DA.

The Lentinus edodes species, known as shiitake, has a history of extensive use in many cuisines in several East Asian countries owing to its unique and pleasant flavor and texture. As international food trade increases, reliable discrimination of geographical origin is becoming increasingly crucial in Korea to identify cheaper imported Chinese shiitake. Herein, stable isotope ratios (i.e., δ13C, δ15N, δ18O, and δ34S) were measured with a stable isotope ratio mass spectrometer, and a geographical discrimination method using orthogonal projection to latent structure-discriminant analysis was developed. The externally validated discrimination method showed excellent predictability (Q2cum = 0.881) and illustrated that δ18O and δ15N were important isotope markers for the geographical discrimination of dried shiitake slices. This study extends the knowledge of geographical differences between China and Korea evidenced by the shiitake isotope signatures, thereby contributing to potential geographical authentication with broader applications for international shiitake markets.

Independent role of low-density lipoprotein cholesterol in subclinical coronary atherosclerosis in the absence of traditional cardiovascular risk factors.

AIMS: Individuals without traditional cardiovascular risk factors (CVRFs) still experience adverse events in clinical practice. This study evaluated the predictors of subclinical coronary atherosclerosis in individuals without traditional CVRFs. METHODS AND RESULTS: A total of 1250 (52.8 ± 6.5 years, 46.9% male) asymptomatic individuals without CVRFs who underwent coronary computed tomographic angiography for a general health examination were analysed. The following were considered as traditional CVRFs: systolic/diastolic blood pressure ≥140/90 mmHg; fasting glucose ≥126 mg/dL; total cholesterol ≥240 mg/dL; low-density lipoprotein cholesterol (LDL-C) ≥160 mg/dL; high-density lipoprotein cholesterol <40 mg/dL; body mass index ≥25.0 kg/m2; current smoking status; and previous medical history of hypertension, diabetes, and dyslipidaemia. Subclinical atherosclerosis, defined as the presence of any coronary plaque, was present in 20.6% cases; the incidences of non-calcified, calcified, and mixed plaque were 9.6%, 12.6%, and 2.6%, respectively. Multivariate regression analysis showed that LDL-C level [odds ratio (OR): 1.008; 95% confidence interval (CI): 1.001-1.015], together with age (OR: 1.101; 95% CI: 1.075-1.128) and male sex (OR: 5.574; 95% CI: 3.310-9.388), was associated with the presence of subclinical atherosclerosis (All P < 0.05). LDL-C level was significantly associated with an increased risk of calcified plaques rather than non-calcified or mixed plaques. CONCLUSION: LDL-C, even at levels currently considered within normal range, is independently associated with the presence of subclinical coronary atherosclerosis in individuals without traditional CVRFs. Our results suggest that a stricter control of LDL-C levels may be necessary for primary prevention in individuals who are conventionally considered healthy.

Cationic Amphipathic Triazines with Potent Anti-bacterial, Anti-inflammatory and Anti-atopic Dermatitis Properties.

The emergence of multi-drug resistant bacteria forces the therapeutic world into a position, where the development of new and alternative kind of antibiotics is highly important. Herein, we report the development of triazine-based amphiphilic small molecular antibacterial agents as mimics of lysine- and arginine-based cationic peptide antibiotics (CPAs). These compounds were screened against a panel of both Gram-positive and Gram-negative bacterial strains. Further, anti-inflammatory evaluation of these compounds led to the identification of four efficient compounds, DG-5, DG-6, DL-5, and DL-6. These compounds displayed significant potency against drug-resistant bacteria, including methicillin-resistant S. aureus (MRSA), multidrug-resistant P. aeruginosa (MDRPA), and vancomycin-resistant E. faecium (VREF). Mechanistic studies, including cytoplasmic membrane depolarization, confocal imaging and flow cytometry suggest that DG-5, DG-6, and DL-5 kill bacteria by targeting bacterial membrane, while DL-6 follows intracellular targeting mechanism. We also demonstrate that these molecules have therapeutic potential by showing the efficiency of DG-5 in preventing the lung inflammation of lipopolysaccharide (LPS)-induced acute lung injury (ALI) mouse model. More interestingly, DL-6 exhibited impressive potency on atopic dermatitis (AD)-like skin lesions in BALB/c mice model by suppressing pro-inflammatory cytokines. Collectively, these results suggest that they can serve a new class of antimicrobial, anti-inflammatory and anti-atopic agents with promising therapeutic potential.

Compound-specific δ13C and δ15N analyses of fatty acids and amino acids for discrimination of organic, pesticide-free, and conventional rice (Oryza sativa L.).

Herein, we improve the procedure for organic rice authentication using compound-specific δ13C and δ15N analyses of fatty acids and amino acids, addressing the increasing demand for accurate methods to confirm organic authenticity. Organic rice (OR) and pesticide-free rice (PFR) featured higher values of δ15Nbulk than conventional rice (CR), whereas the corresponding differences between OR and PFR were insignificant. Additionally, OR, PFR, and CR could be discriminated based on some δ13Camino-acid and δ15Namino-acid values. δ13Cbulk was correlated with most δ13Cfatty-acid (r ≥ 0.596) values, and δ15Nbulk was strongly correlated with most δ15Namino-acid (r ≥ 0.834) values. The first component in the orthogonal projection to latent structure-discriminant analysis model allowed for a clear separation between OR and PFR, and good predictability (Q2Y = 0.506). Thus, the present study improves the reliability of organic authentication when bulk stable isotope ratio analysis alone is insufficient for the accurate discrimination of OR, PFR, and CR.

Fatty Acid- and Amino Acid-Specific Isotope Analysis for Accurate Authentication and Traceability in Organic Milk.

The present study describes compound-specific δ13C and δ15N analyses of fatty acids and amino acids for improving the accurate authentication of organic milk (OM) against conventional milk (CM) collected in Korea. Most δ13Cfatty-acid and δ13Camino-acid values were lower in OM than in CM ( P < 0.05); however, most δ15Namino-acid values displayed weak discriminative power for OM authentication. Higher isotopic fractionation was observed in δ13Cfatty-acid than in δ13Camino-acid and δ15Namino-acid, with fractionation trends differing with individual amino acids. In particular, δ13Clinoleic-acid of -33.5‰ and δ13Cmyristic-acid of -28‰ were determined to be promising year-round threshold values for Korean OM authentication. The δ13Cbulk was highly correlated with δ13CAla ( r = 0.92) and δ13Coleic-acid, trans ( r = 0.77), and strong positive correlations were observed between δ13CVal and δ13CIle ( r = 0.98) and between δ15NThr and δ15NSer ( r = 0.90). Chemometric modeling for OM authentication produced a high quality model ( R2 X = 0.547, R2 Y = 0.865, and Q2 = 0.689) with reliable chemical markers, notably δ13Cmyristic-acid, δ13Clinoleic-acid, and δ13Cstearic-acid. Furthermore, the models developed for seasonal separation in OM ( Q2 = 0.954) and CM ( Q2 = 0.791) were of good quality. Our findings, based on compound-specific isotope data, improve the reliability of OM authentication in cases where bulk stable isotope ratio analysis alone is insufficient. They also provide valuable insight into the control of fraudulent OM labeling in Korea, with potential application in other countries.

Analysis of Selected Phenolic Compounds in Organic, Pesticide-Free, Conventional Rice (Oryza sativa L.) Using LC-ESI-MS/MS.

Rice (Oryza sativa L.) contains generous amounts of carbohydrates, proteins, vitamins, and dietary fibers, in addition to secondary metabolites such as phenols and flavonoids that act as antioxidants. The phenolic compounds detected in rice (organic rice (OR), conventional rice (CR), and pesticide-free rice (PFR)), namely, protocatechuic, gentisic, p-hydroxybenzoic, p-coumaric, ferulic, salicylic, and caffeic acids, are notable free radical scavengers. The sum of these phenolic compounds was found to be higher in PFR, followed by CR and OR (p < 0.0001), when the rice types were classified based on the farming system employed. In addition, significant differences were observed in the p-hydroxybenzoic acid levels for the OR and CR groups compared with the PFR groups (p < 0.01). Furthermore, greater quantities of p-coumaric acid were found in CR-08 and OR-02, although these groups contained overall higher and lower sums of phenolic compounds, respectively. Moreover, significance was observed in the sum of the phenolic compounds, although only small quantities were found in polished rice. Further research is thus required to provide a clearer picture regarding the phenolic profiles of different rice brands.

Nutrition Intervention through Interdisciplinary Medical Treatment in Hospice Patients: From Admission to Death.

The demand for hospice services as well as for 'well-dying' of terminal patients is increasing as patient financial burden is decreasing due to National Health Insurance coverage for hospice care. Hospice institutions utilize interdisciplinary teams comprising doctors, nurses, dietitians, and other health staffs to provide comprehensive patient management. This report examined the nutritional status of a hospice patient from admission to death as well as the nutrition management of this patient in the hospice ward through nutrition interventions performed by a dietitian in the interdisciplinary team. The patient in the present case was a 74-year-old man diagnosed with pancreatic head cancer who died after 26 days of hospice care following transfer from the general ward. During hospice care, the dietitian monitored the patient's nutritional status and performed 8 nutrition interventions, but his oral intake decreased as the patient's symptoms worsened. The average energy intake rates were 30% and 17% of required rates for oral and artificial nutrition, respectively. In line with a report suggesting that the main focus of nutrition in palliative care should be on improving the quality of life and reducing worry in patients, rather than aggressive nutritional management, there is a need for nutrition interventions that are personalized to individual patients by monitoring progress and offering continuous counseling from the time of admission. In addition, further studies such as comparative analysis of nutritional management in Korean hospice ward will be needed for better nutrition management for terminally ill patients.

Metastatic prostate cancer initially presenting as chylothorax: A case report.

Chylothorax is caused by disruption or obstruction of the thoracic duct, which results in leakage of chyle in the pleural space. The most common etiologies are malignancy and trauma. Among the causative malignancies, lymphoma is the most common, followed by primary lung cancer, mediastinal tumors, and other metastatic malignancies. Conversely, prostate cancer has rarely been reported as the cause of chylothorax. We herein report a case of metastatic prostate cancer initially presenting as chylothorax, with disappearance of the pleural effusion after the initiation of androgen deprivation therapy. Moreover, we also discuss the various rare manifestations of metastatic prostate cancer, including chylothorax.

VvpM, an extracellular metalloprotease of Vibrio vulnificus, induces apoptotic death of human cells.

A pathogenic bacterium, Vibrio vulnificus produces various extracellular proteases including the elastolytic metalloprotease, VvpE. In silico analysis of its genome revealed a VvpE-homologous protease, VvpM whose proteolytic activity was abolished by specific inhibitors against metalloproteases. To investigate whether this newly identified protease has pathogenic role in host interaction in addition to proteolytic role, human cell lines were incubated with recombinant VvpM (rVvpM). rVvpM-challenged cells showed typical morphological changes found in cells under apoptosis. Apoptotic cell death was further evidenced by estimating the Annexin V-stained cells, whose proportions were dependent upon the concentrations of rVvpM treated to human cells. To elucidate the signaling pathway for VvpM-induced apoptosis, three MAPKs were tested if their activation were mediated by rVvpM. ERK1/2 was phosphorylated by treatment of rVvpM and rVvpM-induced cell death was blocked by a specific inhibitor against ERK1/2. In rVvpM-treated cells, the cytosolic levels of cytochrome c were increased in a VvpM concentration-dependent manner, while the levels of cytochrome c in mitochondria were decreased. Cell deaths were accompanied by apparent cleavages of procaspases-9 and -3 to the active caspases-9 and -3, respectively. Therefore, this study demonstrates that an extracellular metalloprotease of V. vulnificus, VvpM induces apoptosis of human cells via a pathway consisting of ERK activation, cytochrome c release, and then activation of caspases-9 and -3.

Facilitated intracellular delivery of peptide-guided nanoparticles in tumor tissues.

Macromolecular nanoparticles can extravasate and accumulate within tumor tissues via the passive targeting system, reflecting enhanced permeability and the retention effect. However, the unsatisfactory tumor therapeutic efficacy of the passive-targeting system, attributable to the retention of extravasated nanoparticles in the vicinity of tumor vessels, argues that a new system that facilitates intracellular delivery of nanoparticles within tumors is needed. Here, we developed hydrophobically modified glycol chitosan (HGC) nanoparticles conjugated with interleukin-4 receptor (IL-4R) binding peptides, termed I4R, and tested them in mice bearing IL-4R-positive tumors. These HGC-I4R nanoparticles exhibited enhanced IL-4R-dependent cellular uptake in tumors compared to nonconjugated nanoparticles, leading to better therapeutic and imaging efficacy. We conclude that I4R facilitates and enhances cellular uptake of nanoparticles in tumor tissues. This study suggests that the intracelluar uptake of nanoparticles in tumors is an essential factor to consider in designing nanoparticles for tumor-targeted drug delivery and imaging.

Cell death mediated by Vibrio parahaemolyticus type III secretion system 1 is dependent on ERK1/2 MAPK, but independent of caspases.

Vibrio parahaemolyticus, which causes gastroenteritis, wound infection, and septicemia, has two sets of type III secretion systems (TTSS), TTSS1 and TTSS2. A TTSS1- deficient vcrD1 mutant of V. parahaemolyticus showed an attenuated cytotoxicity against HEp-2 cells, and a significant reduction in mouse lethality, which were both restored by complementation with the intact vcrD1 gene. V. parahaemolyticus also triggered phosphorylation of mitogenactivated protein kinases (MAPKs) including p38 and ERK1/2 in HEp-2 cells. The ability to activate p38 and ERK1/2 was significantly affected in a TTSS1-deficient vcrD1 mutant. Experiments using MAPK inhibitors showed that p38 and ERK1/2 MAPKs are involved in V. parahaemolyticus-induced death of HEp-2 cells. In addition, caspase-3 and caspase-9 were processed into active forms in V. parahaemolyticus-exposed HEp-2 cells, but activation of caspases was not essential for V. parahaemolyticusinduced death of HEp-2 cells, as shown by both annexin V staining and lactate dehydrogenase release assays. We conclude that secreted protein(s) of TTSS1 play an important role in activation of p38 and ERK1/2 in HEp-2 cells that eventually leads to cell death via a caspaseindependent mechanism.