N-Acetylcysteine Alleviates the Progression of Chronic Kidney Disease: A Three-Year Cohort Study.

Background and Objectives: The prevalence of chronic kidney disease (CKD) is approximately 10% of the population in many countries. CKD progresses to end-stage renal disease (ESRD), resulting in adverse outcomes, prolonged hospitalization, and increased healthcare costs. Therefore, reducing CKD progression to ESRD is recognized as an important health issue. Materials and Methods: Data from the study participants with stage 3 to stage 5 CKD (n = 7668) were collected from the National Health Insurance (NHI) program in Taiwan (1 November 2014 to 31 December 2020). CKD patients who had ingested or not ingested N-acetylcysteine (NAC) for three years were divided into the study group (NAC users; n = 165) and the control group (NAC non-users; n = 165) to explore whether NAC use could alleviate CKD progression and reduce the risks associated with hemodialysis in CKD patients. Results: The levels of serum creatinine (SCr) and estimated globular filtration rate (eGFR) were nearly unchanged and/or slightly changed in NAC users, but the SCr levels were slightly increased, and the eGFR levels were significantly decreased in NAC non-users at the six-month interval during the three years. A statistical difference was observed between the two groups for both levels from 12 months to 36 months. The incidence rate of hemodialysis was significantly lower in NAC users than in non-NAC users (4.8% vs. 12.7%, Wald test = 5.947, p = 0.015, OR = 34.9). These results indicated that NAC use may improve renal function of CKD patients by modulating SCr and eGFR and, in turn, reducing the risk of hemodialysis. Conclusions: We investigated whether NAC could be used to reduce CKD progression to ESRD. For the three-year retrospective study, the incidence rate of hemodialysis was significantly lower in NAC users than in non-NAC users via modulating SCr and eGRF levels. NAC use might be a useful clinical approach for reducing CKD progression to ESRD.

Surgical Treatment for Empyema Thoracis: Prognostic Role of Preoperative Transthoracic Echocardiography and Serum Calcium.

BACKGROUND: Empyema is a major cause of mortality and hospitalization. Symptoms include difficulty breathing and chest pain. Calcium plays an essential role in the physiology of the cardiovascular system. However, there is little evidence on the role of echocardiography and the serum calcium levels of patients undergoing video-assisted thoracoscopic surgery (VATS) for empyema. This study aimed to investigate the risk factors for postoperative mortality in patients with empyema who required surgery. METHODS: This single-institution retrospective study compared the outcomes of VATS for thoracic empyema (in terms of survival and mortality) in 122 patients enrolled between July 2015 and June 2019. RESULTS: This study examined patients with thoracic empyema. The majority of the patients were males (100/122, 81.9%). The in-hospital/30-day mortality rate was 10.6% (13 patients). The calcium levels were 7.82 ± 1.17 mg/dL in the survival group and 6.88 ± 1.88 mg/dL in the mortality group (p = 0.032). In the mortality group, the utilization of echocardiography and serum calcium levels independently contributed to the risk prediction more than clinical variables. Patients in our cohort exhibited elevated pulmonary artery systolic pressure (PASP) and hypocalcemia, which were associated with increased postoperative mortality. CONCLUSION: Elevated PASP and calcium levels at the low end of the normal range demonstrated significant prognostic value in predicting mortality in patients with thoracic empyema who required surgical intervention. Recognizing this potential is critical in order to obtain better outcomes.

Omega-3 Fatty Acids Improve Chronic Kidney Disease-Associated Pruritus and Inflammation.

Background and Objectives: Chronic kidney disease-associated pruritus (CKD-aP) is a common symptom in hemodialysis patients. A frequent and intense itching sensation largely torments patients, impacts quality of life outcomes, and it has an independent association with mortality. The objective of this study is to investigate the effects of oral supplementation with omega-3 polyunsaturated fatty acid (omega-3 PUFA) on circulating interleukin-6 (IL-6), cardiometabolic parameters, skin moisturization, and the consequent symptoms of pruritus in hemodialysis patients. Materials and Methods: Volunteers on maintenance hemodialysis with very severe pruritus symptoms were enrolled in this prospective cohort study. Subjects were instructed to consume 1000 mg fish oil once daily for 3 months. Pruritus scoring, skin moisture, plasma IL-6, and cardiometabolic parameters were measured at baseline, and at the first, second, and third month post-supplementation with fish oil for assessment of the clinical significance. Results: A total of 27 patients who had a mean age of 67.33 ± 11.06 years and 3.98 ± 3.23 years on hemodialysis completed the study. Supplementation with omega-3 PUFA significantly decreased IL-6 levels (p < 0.001), but increased the levels of c-reactive protein (CRP) (p < 0.05). Evaluation of the cardiovascular risk showed significant (all p < 0.001) decreases in the total cholesterol (CHO), low-density lipoprotein (LDL), and triglycerides (TG) levels, and an increase in the high-density lipoprotein (HDL) level. A significant decrease in plasma creatinine (CR) was observed (p < 0.001), but the decrease was limited. Supplementation with omega-3 PUFA significantly improved (all p < 0.001) skin hydration on both the face and arms, as well as disease-related symptoms of pruritus. Conclusion: Omega-3 PUFA supplementation improved inflammation, renal function, cardiovascular parameters, dry skin conditions, and the consequent symptoms of pruritus in hemodialysis patients.

A Novel ALDH2 Activator AD-9308 Improves Diastolic and Systolic Myocardial Functions in Streptozotocin-Induced Diabetic Mice.

Diabetes mellitus has reached epidemic proportion worldwide. One of the diabetic complications is cardiomyopathy, characterized by early left ventricular (LV) diastolic dysfunction, followed by development of systolic dysfunction and ventricular dilation at a late stage. The pathogenesis is multifactorial, and there is no effective treatment yet. In recent years, 4-hydroxy-2-nonenal (4-HNE), a toxic aldehyde generated from lipid peroxidation, is implicated in the pathogenesis of cardiovascular diseases. Its high bioreactivity toward proteins results in cellular damage. Mitochondrial aldehyde dehydrogenase 2 (ALDH2) is the major enzyme that detoxifies 4-HNE. The development of small-molecule ALDH2 activator provides an opportunity for treating diabetic cardiomyopathy. This study found that AD-9308, a water-soluble andhighly selective ALDH2 activator, can improve LV diastolic and systolic functions, and wall remodeling in streptozotocin-induced diabetic mice. AD-9308 treatment dose-dependently lowered serum 4-HNE levels and 4-HNE protein adducts in cardiac tissue from diabetic mice, accompanied with ameliorated myocardial fibrosis, inflammation, and apoptosis. Improvements of mitochondrial functions, sarco/endoplasmic reticulumcalcium handling and autophagy regulation were also observed in diabetic mice with AD-9308 treatment. In conclusion, ADLH2 activation effectively ameliorated diabetic cardiomyopathy, which may be mediated through detoxification of 4-HNE. Our findings highlighted the therapeutic potential of ALDH2 activation for treating diabetic cardiomyopathy.

Age Structural Transitions and Copayment Policy Effectiveness: Evidence from Taiwan's National Health Insurance System.

BACKGROUND: Population ageing is a worldwide phenomenon that could influence health policy effectiveness. This research explores the impact of age structural transitions on copayment policy responses under Taiwan's National Health Insurance (NHI) system. METHODS: The time-varying parameter vector autoregressive model was applied to create two measures of the copayment policy effectiveness, and multiple linear regression models were used to verify the nonlinear effect of age structural transitions on copayment policy responses. RESULTS: Our results show that copayment policy effectiveness (in terms of the negative response of medical center outpatient visits to upward adjustments in copayment) is positively correlated with the proportions of the population in two older age groups (aged 55-64 and ≥ 65) and children (age < 15), but negatively correlated with the proportion of the population that makes up most of the workforce (aged 15‒54). These tendencies of age distribution, which influence the responses of medical center outpatient visits to copayment policy changes, predict that copayment policy may have a greater influence on medical center outpatient utilization in an ageing society. CONCLUSIONS: Policymakers should be concerned about the adverse effects of copayment adjustments on the elderly, such as an increasing financial burden and the effect of pricing some elderly patients out of Taiwan's NHI system.

Unmet Supportive Care Needs of Survival Patients with Nasopharyngeal Carcinoma.

This study examined unmet supportive care needs for nasopharyngeal carcinoma (NPC) patients by cancer stage and treatment phase, as well as the factors associated with these unmet needs. At a cancer center in central Taiwan, information on consultations and services patients received at the resource center was described in the service chart. We extracted data available for NPC patients to evaluate their unmet supportive care needs (health information, patient care, treatment, nutritional, psychosocial, and economic) and their association with sex, age, cancer stage, and treatment phase. The 145 NPC patients were 68.3% male, 60.0% less than 50 years old, and 83.5% diagnosed at stages III and IV. The most prevalent unmet need was nutritional (40.7%), followed by psychosocial and patient care, with economic unmet needs the least (4.8%). Women were more likely than men to have patient care unmet needs (32.6% vs. 15.2%). Nutritional unmet need was higher in older patients than in younger ones (83.3% vs. 35.6%), with an adjusted odds ratio (aOR) of 9.39 (95% confidence interval (CI) = 2.17-40.70). Psychosocial unmet needs were higher in younger patients than old patients (34.5% vs. 0%) and in patients interviewed during follow-up period than those at newly diagnosed (55.2% vs. 23.1%). In conclusion, the most commonly reported concern was nutritional unmet needs for NPC patients. Their unmet needs may vary by demographic and disease factors, including patient sex and age, cancer stage, and treatment phase.

Clinical Outcome and Risk Factors for Emergency Department Adult Patients with Thoracic Empyema after Video-Assisted Thoracic Surgical Procedure.

Background: Empyema is a purulent infection of the pleural cavity that is most relevant to parapneumonia effusion. Video-assisted thoracoscopic surgery (VATS) is an option for stage 2 (fibrinopurulent) and stage 3 (organizational). Surgeons may see critically ill patients with pleural empyema who present to the emergency department (ED). The purpose of this work is to investigate the outcomes of ED adult patients with thoracic empyema undergoing a thoracoscopic surgical procedure and to identify possible risk factors for death. Methods: We reviewed retrospectively the clinical characteristics and treatment outcomes of patients with thoracic empyema who received this diagnosis at our center from January 2012 to June 2014. Patients <20 years old were excluded from this study. The prognostic values of age, sex, comorbidities, clinical presentations, location, stage, and laboratory examinations were evaluated. Uni-variable analysis and multi-variable modeling were performed to determine significant risk factors for post-operative death. Results: Seventeen of 160 patients died post-operatively. Two groups (survivors and non-survivors) significantly differed in age (p = 0.013), sex (p = 0.026), comorbidity (p = 0.017), cough (p = 0.024), chest pain (p = 0.016), serum hemoglobin (p = 0.001), and potassium (p = 0.004) levels. Further logistic regression analysis showed statistically significant differences in age, hemoglobin levels, and potassium levels. Conclusion: Among the ED patients with thoracic empyema, older age, lower hemoglobin levels, and higher potassium levels are associated with post-operative death after VATS. These findings underline the importance of careful peri-operative treatment in older patients with signs of empyema when they present to the ED.

Robotic Nipple-Sparing Mastectomy and Immediate Breast Reconstruction with Gel Implant: Technique, Preliminary Results and Patient-Reported Cosmetic Outcome.

BACKGROUND: Experience with application of a robotic surgery platform in the management of breast cancer is limited. The preliminary results of the robotic nipple-sparing mastectomy (R-NSM) and immediate breast reconstruction (IBR) with Gel implant procedure are reported. METHODS: The medical records of patients from a single institution who underwent an R-NSM and IBR with Gel implant procedure for breast cancer during the period March 2017 to February 2018 were assessed. Data on clinicopathologic characteristics, type of surgery, complications, and recurrence were analyzed to determine the effectiveness and oncologic safety of R-NSM. Patient-reported cosmetic outcome results were obtained. RESULTS: A total of 22 patients who received 23 R-NSM and IBR with Gel implant procedures were analyzed. The mean operation time for R-NSM was 118.8 ± 50.6 min, and 74.5 ± 26.6 min for Gel implant reconstruction. Docking time quickly dropped from 20 to 6-8 min, and the time needed to complete R-NSM was usually completed within 100 min after accumulation of case experience. Mean blood loss was 37 ± 38.2 mL, and the positive surgical margin rate was 0%. Three (13%) patients had transit nipple ischemia change, and no total nipple-areolar complex necrosis cases were observed. No local recurrence or mortality was found during a mean 6.9 ± 3.5 months of follow-up. All 22 patients were satisfied with the postoperative aesthetic outcome. CONCLUSION: From our preliminary experience, R-NSM and IBR with Gel implant is a safe procedure, with good cosmetic results, and could be a promising new technique for breast cancer patients indicated for mastectomy.

Robotic Nipple-sparing Mastectomy and Immediate Breast Reconstruction with Gel Implant.

BACKGROUND: The experience of application of robotic surgery platform in the management of breast cancer was limited. The preliminary experience and results of robotic nipple-sparing mastectomy (R-NSM) and immediate breast reconstruction (IBR) with Gel implant was reported. METHODS: The medical records of patients who underwent R-NSM and IBR with Gel implant for breast cancer during the period March 2017 to October 2017 were collected from single institution. Data on clinicopathologic characteristics, type of surgery, method of breast reconstruction, complications, and recurrence were analyzed to determine the effectiveness and oncologic safety of R-NSM. Patients' oriented cosmetic outcome report was also obtained. RESULTS: A total of 15 patients was analyzed, and the mean age of them was 46.5 ± 10.0 years. The pathologic stage was 30.8% ductal carcinoma in situ, 30.8% stage I, 30.8% stage II, and 7.7% stage III. The mean operation time was 282.8 ± 70.4 minutes, and mean hospital stay was 6.7 ± 1.2 days. The positive surgical margin rate was 0%. One patient suffered from delayed axillary wound healing. Two patients (13.3%) with transit nipple ischemia change, but no total nipple areolar complex necrosis case was observed. No local recurrence, distant metastasis, or case mortality was found during mean 6.3 ± 2.1 months follow-up. All 15 patients were satisfied with the postoperative aesthetic outcome. CONCLUSION: From our preliminary experience, R-NSM and IBR with Gel implant is a safe procedure, with good cosmetic results, and could be a promising new technique for breast cancer patients indicated for mastectomy.

Technique for single axillary incision robotic assisted quadrantectomy and immediate partial breast reconstruction with robotic latissimus dorsi flap harvest for breast cancer: A case report.

RATIONALE: The clinical application of robotic surgery in breast conserving surgery or volume replacement with robotic latissimus dorsi flap harvest (RLDFH) has been rarely reported. In this study, we report the preliminary experience and clinical outcome of robotic assisted quadrantectomy (RAQ) and immediate partial breast reconstruction (IPBR) with RLDFH. PATIENT CONCERN: Decreasing and avoid back scar length after latissimus dorsi flap harvest. DIAGNOSES: One 28 years old female with left breast cancer underwent RAQ and IPBR with RLDFH. Initially, she was diagnosed with left breast infiltrating carcinoma that was clinical stage T3N1M0 and triple negative. INTERVENTIONS: Neoadjuvant chemotherapy consisting of 4 cycles of epirubicin and cyclophosphamide followed by 4 cycles of docetaxel was performed. Breast magnetic resonance imaging showed residual breast cancer about 4.5 cm over the left upper outer quadrant of the breast. Sentinel lymph node biopsy showed no lymph node metastasis. RAQ, which took 82 minutes, was performed first, and the resected breast specimen's weight was 203 gm. She received IPBR with RLDFH, which took 97 minutes. OUTCOMES: The overall blood loss was 40 mL. The final pathology result was ypT2 (4.2 cm)N0 (sn0/3)M0 and stage IIA. The resection margin was free of tumors. The post-operative recovery was smooth except for seroma formation over the back, which was relieved after repeated aspiration at an outpatient clinic. The patient was satisfied with the post-operative scar and aesthetic outcome. No local recurrence, distant metastasis or case mortality was found during 5 months of follow-up. LESSONS: RAQ and IPBR with RLDFH is a safe alternative for small-to-medium-breast-size women with breast cancer who desire breast conservation and are indicated for volume replacement with autologous latissimus dorsi flap.

Single-Axillary-Incision Endoscopic-Assisted Hybrid Technique for Nipple-Sparing Mastectomy: Technique, Preliminary Results, and Patient-Reported Cosmetic Outcome from Preliminary 50 Procedures.

BACKGROUND: A new hybrid technique for single-axillary-incision endoscopic-assisted nipple-sparing mastectomy (E-NSM) was introduced. Preliminary results are reported. METHODS: Patients who received single-axillary-incision E-NSM from August 2013 to August 2017 were searched from a single institution. Data were analyzed to determine the effectiveness and oncologic safety of single-axillary-incision E-NSM. Patient-oriented cosmetic outcome report was also obtained. RESULTS: During the study period, a total of 50 E-NSM with single-incision procedures were performed in 41 female patients with breast cancer, including 11 (26.8%) patients with bilateral disease. Their mean age was 45.3 ± 8.4 years. The mean size of tumors encountered during the 50 single-incision E-NSM procedures was 2.3 ± 1.8 (0.1-7.3) cm for invasive tumors and 2.6 ± 1.7 (0.2-5.7) cm for carcinoma in situ lesions. Six (12%) of those tumors were multifocal/multicentric. Lymph node metastasis was found during 12% of the procedures. Forty-five (90%) received immediate breast reconstruction with gel implant. Mean operating time was 244.3 ± 82.8 min. The overall complication rate was 6%, and no total nipple necrosis or implant loss was observed. No locoregional recurrence or distant metastasis was found during mean follow-up of 21.6 months. About 94.4% of patients were satisfied with the postoperative scar location and wound length. All patients who responded would choose the same operation again. CONCLUSIONS: The proposed single-axillary-incision endoscopic hybrid technique for nipple-sparing mastectomy was a safe procedure with low morbidity and associated with high patient satisfaction.

The Effects of White Noise on Agitated Behaviors, Mental Status, and Activities of Daily Living in Older Adults With Dementia.

BACKGROUND: The aging of society is a global trend, and care of older adults with dementia is an urgent challenge. As dementia progresses, patients exhibit negative emotions, memory disorders, sleep disorders, and agitated behavior. Agitated behavior is one of the most difficult problems for family caregivers and healthcare providers to handle when caring for older adults with dementia. PURPOSE: The aim of this study was to investigate the effectiveness of white noise in improving agitated behavior, mental status, and activities of daily living in older adults with dementia. METHODS: An experimental research design was used to study elderly participants two times (pretest and posttest). Six dementia care centers in central and southern Taiwan were targeted to recruit participants. There were 63 participants: 28 were in the experimental group, and 35 were in the comparison group. Experimental group participants received 20 minutes of white noise consisting of ocean, rain, wind, and running water sounds between 4 and 5 P.M. daily over a period of 4 weeks. The comparison group received routine care. Questionnaires were completed, and observations of agitated behaviors were collected before and after the intervention. RESULTS: Agitated behavior in the experimental group improved significantly between pretest and posttest. Furthermore, posttest scores on the Mini-Mental Status Examination and Barthel Index were slightly better for this group than at pretest. However, the experimental group registered no significant difference in mental status or activities of daily living at posttest. For the comparison group, agitated behavior was unchanged between pretest and posttest. CONCLUSIONS: The results of this study support white noise as a simple, convenient, and noninvasive intervention that improves agitated behavior in older adults with dementia. These results may provide a reference for related healthcare providers, educators, and administrators who care for older adults with dementia.

Influence of differing nitrate and nitrogen availability on flowering control in Arabidopsis.

Nitrogen, an essential macronutrient for plants, regulates many aspects of plant growth and development. Nitrate is one of the major forms of nitrogen taken up by plants from the soil. Nitrate and nitrogen have been reported to regulate flowering; while some studies have shown that lower nitrate/nitrogen promoted flowering, others have reported the opposite trend. To elucidate how nitrate/nitrogen affects flowering, we reviewed the existing literature and conducted experiments to examine flowering time under a wide range of nitrate concentrations using two growth systems. From the literature review and our experiments, we established that differing nitrate availability results in a U-shaped flowering curve, with an optimal concentration of nitrate facilitating flowering and concentrations above or below this optimal concentration delaying flowering. The role of nitrate and nitrogen in regulating flowering has been elucidated by several transcriptomic and mutant studies, which have suggested close interactions between nitrate/nitrogen, phosphate, the circadian clock, photosynthesis, and, potentially, hormones. We discuss several possible molecular mechanisms underlying the U-shaped flowering response.

A SimpleProbe(®) real-time PCR assay for differentiating the cytochrome b M121I mutation in clinical specimens from dogs infected with Babesia gibsoni.

Babesia gibsoni (B. gibsoni) causes a canine tick-borne disease worldwide. The substitution of methionine with isoleucine (M121I) in the cytochrome b (CYTb) gene of B. gibsoni was identified as being associated with atovaquone resistance. Rapid identification of the drug-resistant strain is required to select a more effective combination of drugs, e.g., from atovaquone and azithromycin (AA) to clindamycin, diminazene, and imidocarb (CDI) combination. A SimpleProbe(®) real-time PCR assay was designed to detect the single nucleotide polymorphism at nucleotide 363 in CYTb gene of B. gibsoni and the sensitivity and specificity were evaluated by comparing the results from the conventional DNA sequencing method. Eighty-nine clinical blood samples were collected and analyzed in parallel with the SimpleProbe(®) assay and DNA sequencing. The assay identified 50 of 54 nt363G samples and had a sensitivity of 92.6% and a specificity of 100%. Thirty nt363T samples were correctly identified, as well, with a sensitivity of 100% and a specificity of 73.2%. However, this assay identified only one of 17 nt363A samples; the other 16 samples were misidentified as nt363T. The sensitivity of the nt363A identification was only 5.9%, and the specificity was 100%. When detecting the M121I mutation, 42 of 42 mutant samples were identified, with a sensitivity of 100%, and 45 of 47 wild type samples were identified, with a specificity of 95.7%. In conclusion, the SimpleProbe(®) assay could be used to detect the M121I mutation of the B. gibsoni CYTb from clinical specimens. This assay provides a reliable and sensitive tool for differentiating between the atovaquone-resistant strain and the non-resistant strain.

Regulatory expression of MMP-8/MMP-9 and inhibition of proliferation, migration and invasion in human lung cancer A549 cells in the presence of HGF variants.

Hepatocyte growth factor (HGF), a potent cytokine of mesenchymal origin, exhibits polytrophic physiological responses, including proliferation, migration, and invasion, in a wide variety of cells. Although it is known that inhibition of the responses by HGF variants was via signal transducers and activators of the transcription pathway, the mechanisms of action of the variants involved in the production of matrix metalloproteinases (MMPs) were not clearly understood. Thus, recombinant HGF variants, NK1, NK2, NK3, and NK4 were topically applied to assays for proliferation, migration, invasion, and expression of MMPs in the human lung cancer cell line A549 and compared to that of control medium and a glutathione-s-transferase control. Results showed that these recombinant HGF variants significantly inhibited proliferation, migration, and invasion of A549 at >4 nM, downregulated expression of MMP-9, and upregulated expression of MMP-8. The study clearly suggests that binding of the HGF variants to the cell surface c-Met resulted in the downregulation of MMP-9, and upregulation of MMP-8 protein expressions might be key molecular signals against proliferation, migration, and invasion of A549 cells.

In vivo relevance of substrate recognition function of major Arabidopsis ubiquitin receptors.

Ubiquitylation marks proteins for destruction by the 26S proteasome. These signals are deciphered and targeted by distinct direct and indirect pathways involving a set of evolutionarily conserved ubiquitin receptors. Although biochemical and structural studies have revealed the mechanistic complexity of these substrate recognition pathways, conclusive evidence of the in vivo relevance of their substrate recognition function is currently not available. We recently showed that the structural elements involved in substrate recognition are not responsible for the important roles of the ubiquitin receptor RPN10 in vegetative and reproductive growth or for the abundance of the two-capped proteasomes (RP2-CP). Moreover, Arabidopsis plants subjected to severe knockdown or knockout any of the major ubiquitin receptors displayed wild-type phenotypes. Our results clearly suggest a functional redundancy of the major Arabidopsis ubiquitin receptors, and this evolved multiplicity is probably used to secure the substrates delivery. Based on the reduced abundance of RP2-CP in rpn10-2 and a role of RPN10 in lid-base association, a structural role of RPN10 in 26S proteasome stability is likely to be more relevant in vivo. Further efforts using structural and functional analyses in higher-order mutants to identify the specific biological functions of substrate recognition for the major Arabidopsis ubiquitin receptors are described here.

Glutathione-S-transferase enhances proliferation-migration and protects against shikonin-induced cell death in breast cancer cells.

Glutathione-S-transferase (GST) is a cytoplasmic protein responsible for detoxification, but the effect of the enzyme on cell biological events, including proliferation and migration, has never been reported. Thus, we evaluated the detoxification effect of in vitro-applied GST on cancer cell proliferation and migration. Assays for proliferation and migration of human breast cancer cells in the presence of GST were carried out. Binding of GST on the surface of the cancer cells was studied by flow cytometry. Detoxification through GST pathway was studied in the presence of shikonin. The effective dosage of GST in enhancement of cell proliferation was 10-50 nM, and the cell migration could be significantly enhanced after 6 hours in the presence of 2-50 nM GST. Therefore, overall cell proliferation and migration could be enhanced in the presence of 10nM or greater concentration of GST, and 15 µM shikonin-induced toxification of the cancer cells could be neutralized by 1.0 µM GST. Flow cytometry showed that GST directly bound to the surface of the cancer cells, and this was confirmed by fluorescence confocal microscopic observation. It is concluded that human class π-GST enhances proliferation and migration of human breast cancer cells by means of direct binding to the cell surface and maintaining cell viability by detoxification.

The defective proteasome but not substrate recognition function is responsible for the null phenotypes of the Arabidopsis proteasome subunit RPN10.

Ubiquitylated substrate recognition during ubiquitin/proteasome-mediated proteolysis (UPP) is mediated directly by the proteasome subunits RPN10 and RPN13 and indirectly by ubiquitin-like (UBL) and ubiquitin-associated (UBA) domain-containing factors. To dissect the complexity and functional roles of UPP substrate recognition in Arabidopsis thaliana, potential UPP substrate receptors were characterized. RPN10 and members of the UBL-UBA-containing RAD23 and DSK2 families displayed strong affinities for Lys-48-linked ubiquitin chains (the major UPP signals), indicating that they are involved in ubiquitylated substrate recognition. Additionally, RPN10 uses distinct interfaces as primary proteasomal docking sites for RAD23s and DSK2s. Analyses of T-DNA insertion knockout or RNA interference knockdown mutants of potential UPP ubiquitin receptors, including RPN10, RPN13, RAD23a-d, DSK2a-b, DDI1, and NUB1, demonstrated that only the RPN10 mutant gave clear phenotypes. The null rpn10-2 showed decreased double-capped proteasomes, increased 20S core complexes, and pleiotropic vegetative and reproductive growth phenotypes. Surprisingly, the observed rpn10-2 phenotypes were rescued by a RPN10 variant defective in substrate recognition, indicating that the defectiveness of RPN10 in proteasome but not substrate recognition function is responsible for the null phenotypes. Our results suggest that redundant recognition pathways likely are used in Arabidopsis to target ubiquitylated substrates for proteasomal degradation and that their specific roles in vivo require further examination.

Infection of Schistosomiasis japanicum is likely to enhance proliferation and migration of human breast cancer cells: mechanism of action of differential expression of MMP2 and MMP9.

OBJECTIVE: To study whether the infection of Schistosomiasis japanicum (S. japanicum) is related to enhanced proliferation and migration of cancer cells, and the molecular mechanism pertains to cancer cell metastasis in human host. METHODS: The gene of S. japanicum glutathione transferase (sjGST) cloned from S. japanicum was expressed, purified and applied in a series of assays to explore the effect of sjGST on proliferation and migration of MDA-MB-435S, and the expression of MMP2 and MMP9. Immunofluorescence assay for the binding of sjGST to MDA-MB-435S was also carried out. RESULTS: Results showed that sjGST enhanced proliferation and migration in human breast cancer cell MDA-MB-435S signifycantly at 50-200 nM, but did not enhance them in human lung cancer cell A549. Immunofluorescence assay for the binding of sjGST to MDA-MB-435S and A549 showed that GST was readily bound to the breast cancer cells, but showed almost no binding to human lung cancer cells. The assays for gelatinase activity showed that both MMP2 and MMP9 activities were increased significantly in the presence of sjGST (50-200 nM) in MDA-MB-435S, but they were not significant in A549. CONCLUSIONS: Our current results show strongly that S. japanicum GST binds to MDA-MB-435S probably via its receptor, and enhances proliferation and migration of the cancer cells by up-regulatory expression of MMP2 and MMP9.