Study of the N=32 and N=34 Shell Gap for Ti and V by the First High-Precision Multireflection Time-of-Flight Mass Measurements at BigRIPS-SLOWRI.

The atomic masses of ^{55}Sc, ^{56,58}Ti, and ^{56-59}V have been determined using the high-precision multireflection time-of-flight technique. The radioisotopes have been produced at RIKEN's Radioactive Isotope Beam Factory (RIBF) and delivered to the novel designed gas cell and multireflection system, which has been recently commissioned downstream of the ZeroDegree spectrometer following the BigRIPS separator. For ^{56,58}Ti and ^{56-59}V, the mass uncertainties have been reduced down to the order of 10 keV, shedding new light on the N=34 shell effect in Ti and V isotopes by the first high-precision mass measurements of the critical species ^{58}Ti and ^{59}V. With the new precision achieved, we reveal the nonexistence of the N=34 empirical two-neutron shell gaps for Ti and V, and the enhanced energy gap above the occupied νp_{3/2} orbit is identified as a feature unique to Ca. We perform new Monte Carlo shell model calculations including the νd_{5/2} and νg_{9/2} orbits and compare the results with conventional shell model calculations, which exclude the νg_{9/2} and the νd_{5/2} orbits. The comparison indicates that the shell gap reduction in Ti is related to a partial occupation of the higher orbitals for the outer two valence neutrons at N=34.

[Research and development of new traditional Chinese medicine drugs for certain syndromes based on "theoretical innovation"].

Compound prescription of traditional Chinese medicine(TCM), the main form in prevention and treatment of diseases in TCM, has unique advantages in medical practice. New TCM drugs for certain syndromes serve as an important carrier of inheritance and innovation in TCM. The research and development of new TCM drugs for certain syndromes should be based on the theory of TCM, guided by clinical value, and based on the principle of quality, safety and effectiveness. Through the innovative understanding of disease development rules, clinicians carry out academic theory innovation to guide clinical practice, aiming to effectively promote academic innovation and the development of new TCM drugs for certain syndromes. In this paper, we expounded some understanding of the innovation system of TCM, and analyzed the research value of new TCM drugs for certain syndromes. Based on theoretical innovation, the overall research model was preliminarily put forward. Subsequently, a concrete discussion from three aspects, including pharmaco-logy and toxicology, pharmacy and clinic research, was made on the specific process and existing problems of new drug research and development of TCM. Our research group attempts to establish a new drug innovation ecosystem for TCM syndrome, with the purpose of providing reference for other researchers.

[A study of bacterial distribution and drug resistance in skin and soft tissue infection].

Objective: To investigate pathogenic bacteria and drug resistance in the patients with skin and soft tissue infection in order to provide the scientific evidences for clinical reasonable use of antibiotics. Methods: A retrospective analysis was performed on patients with skin and soft tissue infections in Department of Dermatology, Peking University Third Hospital from January 2012 to December 2017. Pus, secretions, skin lesions, urine, throat swabs, and alveolar lavage fluid were collected for bacterial culture, bacterial species were identified by VITEK2 Compact system and BD-Bruker MALDI Biotyper system. Drug resistance was detected by K-B agar diffusion method recommended by CLSI. Results: A total of 392 strains of bacteria were isolated from 327 patients distributed in 21 genus and 56 species, of which 225 were gram-positive cocci (57.40%), 114 were gram-negative rods (29.08%), 46 were gram-positive rods (11.73%), and 7 were gram-negative cocci (1.79%). The top 3 bacteria were Staphylococcus aureus 91(23.21%), Staphylococcus epidermidis 42 (10.71%), and Pseudomonas aeruginosa 24 (6.12%). Staphylococcus had a high rate of resistance to penicillin and erythromycin (>50%). Gram-negative rods were resistant to ampicillin (86.1%), and also had certain resistance to most second-generation and some third-generation cephalosporin (about 50%). There was no significant change in the drug resistance rate of MRSA compared to MSSA. Only the resistance rate to tetracycline was statistically different (P<0.05). Conclusion: The emergence of drug-resistant strains is an important factor leading to refractory infections. There are a wide range of pathogenic bacteria species among the skin and soft tissue infection patients, and antimicrobial drugs should be chosen wisely according to drug sensitivity.

CEACAM 6, a novel marker for the diagnosis of Barrett's esophagus.

Barrett's esophagus (BE) is a premalignant condition associated with the development of esophageal adenocarcinoma (EAC). Despite the low risk of progression to EAC, evidence highlights the notably poor survival rates of this malignancy. The mainstay form of diagnosis of BE is endoscopy and biopsy sampling. However, research emphasizes limitations with regards to the histological detection of BE and associated dysplasia. The aim of this study is to evaluate the clinical significance of CEACAM6 as a potential biomarker for the diagnosis of BE and beyond. Retrospective tissue samples were obtained from columnar lined esophagus without goblet cells (n = 27), BE (n = 18), BE associated dysplasia (n = 16), and EAC (n = 24). Standardized immunohistochemistry for CEACAM6 was performed followed by quantitative staining analysis. Statistical analysis across the BE spectrum for CEACAM6 was undertaken and a P value <0.05 was considered significant. CEACAM6 expression increased from columnar lined epithelium (CLE) to BE with a subsequent decrease to dysplasia and adenocarcinoma. The expression of CEACAM6 was significant from CLE to BE at p 0.001, CLE to dysplasia at p 0.001, BE to dysplasia at p 0.006, CLE to adenocarcinoma at p 0.001 and BE to adenocarcinoma at p 0.001. There was no significant difference in expression between dysplasia and adenocarcinoma (P = 0.15). Our findings highlight the increasing expression of CEACAM6 from CLE to BE with a subsequent decrease to dysplasia and adenocarcinoma. In view of this, we advocate the utilization of this marker for the enhanced diagnosis of BE and for the distinction of BE and dysplasia.

Predictive value of vascular endothelial growth factor polymorphisms on the risk of renal cell carcinomas.

We conducted a case-control study in a Chinese population to assess whether 5 common single-nucleotide polymorphisms in the vascular endothelial growth factor gene (VEGF) affect the risk of renal cell carcinoma (RCC). The study population included 266 RCC patients who were newly diagnosed and histologically confirmed to have RCC as well as 532 cancer-free controls. Genotyping of VEGF -2578C/A, -1156G/A, +1612G/A, +936C/T, and -634G/C was conducted by polymerase chain reaction-restriction fragment length polymorphism. RCC patients were more likely to have higher body mass index, and have a habit of tobacco smoking as well as suffer from diabetes. Conditional logistic regression analyses showed that individuals with the AA genotype and A allele of -2578C/A significantly increased the risk of RCC when compared with the CC genotype. Individuals carrying the CT and TT geno-types of +936C/T were correlated with an increased risk of RCC compared to the CC genotype. The T allele of +936C/T was associated with an increased risk of RCC. The -2578C/A and +936C/T polymorphisms in the VEGF gene may play a role in the etiology of RCC.

Biological differences and molecular mechanism of different peripheral nerves after injury.

OBJECTIVE: The aim of the study was to analyze the apoptosis of neurons and the differences in expression of Bcl-2 and Bax protein in the neurons in the corresponding spinal cord segment after the repair of the tibial nerve (TN) and common peroneal nerve (CPN) in rats. MATERIALS AND METHODS: 126 healthy male Sprague-Dawley (SD) rats aged 7-8 weeks were randomly divided into group A (control group), group B (TN was cut and sutured), and group C (CPN was cut and sutured), with 42 rats in each group. The spinal cord tissues of rats in different groups were stained with hematoxylin-eosin (HE) on the 1st, 3rd, 7th, 14th, 21st, and 28th day after surgery; the number of neurons in anterior horn of spinal cord, axon density (AD), axon passage rate (APR), and recovery rate (RR) of muscle cell cross-sectional area (MCCA) were calculated; and differences in the expression of Bcl-2 and Bax proteins in the three groups of rats were analyzed by immunohistochemistry. RESULTS: The results showed that there was no statistically significant difference in the muscle wet weight (MWW) RR of the three groups of rats on the 14th day after the surgery (p>0.05), and the MWW RRs of rats in groups B and C were higher at the 28th day after surgery in contrast to group A (p<0.05). The number of motor neurons in the anterior horn of spinal cord in group B was higher than that in group C at the 3rd, 7th, 14th, and 21st day after surgery (p<0.05); the MWW RR, MCCA, and CSARR of rats in group B were lower than those in group C (p<0.05); the proximal AD, distal AD, and APR in group B were higher than those of group C on the 14th and 28th day after the surgery (p<0.05); and there were no positive staining results in the spinal cord tissue of rats in group A after staining. The expressions of Bcl-2 and Bax in group B were higher observably than the expressions in group C (p<0.05), which indicated that the recovery ability of TN was stronger than that of the CPN; the expression of Bcl-2 and Bax in TN was notably higher than that of the CPN. CONCLUSIONS: The expression of Bcl-2 and Bax was related to cell apoptosis and nerve regeneration after nerve injury. It provided a reference basis for clinical diagnosis and treatment of peripheral nerves.

Increased survival and reduced renal injury in MRL/lpr mice treated with a human Fcγ receptor II (CD32) peptide.

Systemic lupus erythematosus (SLE) is a multisystem chronic inflammatory disease affecting many organs. The deposition in kidney tissue of immune complexes and their interaction with macrophages is thought to trigger the inflammatory response leading to glomerulonephritis. It has been demonstrated that inhibition of this interaction in murine models can alleviate the disease. Six synthetic peptides were derived from the membrane-proximal extracellular domain (EC2) of human Fcγ receptor II (huFcγRII). Of these, one peptide, huRII6, was shown to be a potent competitive inhibitor of IgG binding to recombinant FcγRII in vitro. To examine the possible therapeutic impact of huRII6 in vivo, this peptide, or a control, was given by subcutaneous injection to female MRL/lpr mice from weeks 7 to 36, resulting in an enhanced survival rate compared with control-treated animals and a reduction of proteinuria. Histopathological examination of the kidneys showed a reduction in deposition of immune complexes and preservation of structure. Such a functional peptide should prove useful for examining the role of IgG-FcγR interactions in experimental models of disease and may provide for the development of FcR-targeting drugs to treat autoimmune disorders.

Confirmation of LRRK2 S1647T variant as a risk factor for Parkinson's disease in southern China.

BACKGROUND: Leucine-rich repeat kinase 2 (LRRK2) S1647T has been identified as a risk variant for Parkinson's disease (PD) in Han Chinese. METHODS: To replicate the association of LRRK2 S1647T with risk of PD, we conducted a case-control study of this variant involving 406 PD subjects and 412 controls from southern mainland China. RESULTS: The results showed that the frequency of A allele was higher in patients with PD (OR=1.238, 95% CI: 1.015-1.510, P=0.035) compared to controls. In a multivariate logistic regression analysis with the disease group (patients with PD vs. controls) as the dependent variable and genotype as an independent factor adjusting for the effect of age and gender, the homozygous S1647T genotype (AA) was associated with an increased risk of PD (OR=1.815, 95% CI:1.270-2.594, P=0.001). The pooled analysis of present data and the data from the previous work demonstrated that the frequency of A allele was higher in patients with PD (OR=1.2, 95% CI: 1.09-1.32, P<0.0001). CONCLUSIONS: LRRK2 S1647T increases the risk of Parkinson's disease in southern China.

[Exploring the therapeutic mechanism of quercetin for heart failure based on network pharmacology and molecular docking].

OBJECTIVE: To investigate the molecular mechanism of quercetin in the treatment of heart failure (HF) based on network pharmacology and molecular docking. METHODS: Quercetin and HF-related targets were obtained using TCMSP, PharmMapper, CTD and GeneCards databases, and quercetin-HF intersection targets were obtained through the online website Venn; the protein interaction network was constructed and imported into Cytoscape 3.7.2 to identify the core targets of quercetin in the treatment of HF.GO and KEGG pathway enrichment analyses were performed using R package, and molecular docking was performed using Auto Dock Vina.The protein levels of AKT1, phospho-AKT(Ser473), eNOS, MMP9, and caspase-3 in quercetin-treated HF cell models were detected using protein immunoblotting. RESULTS: We identified 80 quercetin-HF intersectional targets (AKT1, CASP3, MAPK1, MMP9, and MAPK8) and 5 core targets of quercetin for treatment of HF.GO analysis suggested that the therapeutic effect of quercetin for HF was mediated mainly by such biological processes as responses to peptide hormones, phosphatidylinositol-mediated signalling, responses to lipopolysaccharides, responses to molecules of bacterial origin and regulation of inflammatory responses.KEGG pathway enrichment analysis identified lipid and atherosclerosis pathway, proteoglycans in cancer, PI3K-AKT signaling pathway, diabetic cardiomyopathy and MAPK signaling pathway as the most significantly enriched signaling pathways.Molecular docking showed a good binding activity of quercetin to the 5 core targets.The results of protein immunoblotting showed that 100 µmol/L quercetin significantly reduced AKT1, phospho-AKT (Ser473), eNOS, MMP9 and caspase-3 levels in the cell models of HF (P < 0.01). CONCLUSION: Quercetin improves the pathological changes in HF possibly by regulating the AKT1-eNOS-MMP9 pathway to inhibit cell apoptosis.

[RIP1/RIP3-MLKL signaling pathway correlates with occurrence, progression and prognosis of chronic heart failure].

OBJECTIVE: To detect plasma levels of receptor-interacting protein kinase 1 (RIP1), RIP3 and mixed lineage kinase domain-like protein (MLKL) in patients with chronic heart failure and explore the expression pattern of programmed necrosis signaling pathway RIP1/RIP3-MLKL in the progression of heart failure. METHODS: The patients with chronic heart failure (NYHA class Ⅱ-Ⅳ) admitted in our hospital between February, 2020 and March, 2021 were prospectively enrolled in this study, with 21 healthy volunteers as the control group. The enrolled patients included 20 with grade Ⅱ, 33 with grade Ⅲ, and 43 with grade Ⅳ cardiac function. Fasting venous blood was collected from all the participants for detecting plasma levels of RIP1, RIP3, and MLKL and protein expressions of RIP1/RIP3-MLKL pathway using enzyme-linked immunosorbent assay (ELISA) and Western blotting. The patients with grade Ⅳ cardiac function were followed up for 5 months to evaluate the clinical prognostic indicators. RESULTS: Compared with the healthy volunteers, the patients with grade Ⅱ, Ⅲ and Ⅳ cardiac function had significantly increased plasma levels of RIP1, RIP3, and MLKL (P < 0.01), and their levels were significantly higher in grade Ⅲ/Ⅳ patients than in those with grade Ⅱ cardiac function (P < 0.01); the plasma levels of RIP1 and MLKL were significantly higher in grade Ⅳ patients than in grade Ⅲ patients (P < 0.05). The results of Western blotting also showed increased expressions of the proteins in the RIP1/RIP3-MLKL pathway in patients with heart failure. Pearson correlation analysis suggested that in patients with heart failure, the expression levels of RIP1, RIP3, and MLKL were positively correlated with SCR, AST, LVEDD and NT-proBNP (P < 0.05). Follow-up study of the patients with grade Ⅳ cardiac function showed that higher expression levels of RIP1/RIP3-MLKL were associated with a poorer prognosis of the patients. CONCLUSION: The expressions of RIP1, RIP3 and MLKL are significantly upregulated in patients with heart failure in positive correlation with the severity of the disease condition, and the activation of the RIP1/RIP3-MLKL signaling pathway may contribute to the occurrence, development and prognosis of chronic heart failure.

Investigation on ultrasonic cavitation erosion of TiMo and TiNb alloys in sulfuric acid solution.

Two kinds of Ti-alloys, i.e., TiMo and TiNb alloys are manufactured in this paper, and their ultrasonic cavitation erosion behaviors in 0.1 M H2SO4 solution are evaluated by the mean depth erosion (MDE), SEM and white light photograph. The results show that MDE of TiMo and TiNb alloys obviously increase with increasing the cavitation erosion time, however, they evidently decrease with the increment of Mo or Nb content at each fixed cavitation erosion time, and even some large blank areas (uneroded areas) still exist on the sample surface after ultrasonic cavitation erosion for 2 h in the case of Ti10Mo and Ti20Nb samples, implying the enhanced anti-cavitation erosion property of Ti-alloy by adding Mo or Nb element. The MDE of Ti10Mo or Ti20Nb sample is lower than that of TC4 sample in the case of each cavitation erosion time, indicating the better cavitation erosion resistance of of Ti10Mo or Ti20Nb sample. The influences of Mo and Nb on the passivity of TiMo and TiNb alloys during the ultrasonic cavitation erosion are detected by potentiodynamic curves. The results display that Ti, TC4, TixMo (x = 1, 5, 10) and TixNb (x = 5, 10, 20) samples are all almost in the passive state within the potential region from 0VSCE to 1.5VSCE during ultrasonic cavitation erosion, and the passive current density evidently decreases with increasing Mo or Nb content, indicating the enhanced passive characteristic by adding Mo or Nb alloys during the ultrasonic cavitation erosion.

Association between sarcopenia and depressive mood in urban-dwelling older adults: A cross-sectional study.

AIM: The aim of the present study was to examine the relationship between depressive mood and diagnostic components of sarcopenia. METHODS: The study used baseline data of participants in the Toyota Prevention Intervention for Cognitive Decline and Sarcopenia study. Participants in this cross-sectional study were 432 older adults (46.5% women, mean age 72.5 ± 4.7 years). We defined sarcopenia using the diagnostic algorithm recommended by the Asian Working Group for Sarcopenia, and all participants were classified into a sarcopenia or healthy control group. The skeletal muscle mass was measured by bioelectrical impedance. Depressive mood was assessed using the Geriatric Depression Scale-15 (range 0-15). RESULTS: Among the 432 participants, 9.5% were classified as having sarcopenia. The mean ± SD Geriatric Depression Scale-15 scores in the control and sarcopenia groups were significantly different at 3.9 ± 2.8 and 5.3 ± 3.3, respectively (P = 0.003). Furthermore, depressive mood was significantly more prevalent in the sarcopenia group (P = 0.011). Multiple linear regression analysis showed that the Geriatric Depression Scale score was associated with grip strength (ß = -0.23, P = 0.004) and walking speed (ß = -0.15, P = 0.006), but not skeletal muscle mass index (ß = -0.16, P = 0.142), after controlling for demographic factors, chronic diseases, inflammatory markers and physical activity. CONCLUSIONS: Sarcopenia was associated with depressive mood. In terms of the diagnostic components of sarcopenia, depressive mood was not associated with decreased muscle mass, but was associated with low muscle strength and low physical performance. Geriatr Gerontol Int 2019; 19: 508-512.

Objectively measured physical activity and cognitive function in urban-dwelling older adults.

AIM: Physical activity (PA) and cognition have reportedly been associated with each other. However, it remains to be elucidated what intensities of PA are most strongly associated with cognition. In the current study, we aimed to determine the association between the intensities of objectively measured PA and cognitive function. METHODS: The present study was a cross-sectional analysis of the data obtained at registration in a randomized control trial in Toyota, Japan. Participants were community-dwelling older adults who had cognitive complaints. A battery of neuropsychological and physical assessments was carried out. Daily PA data were collected with the activity monitor. PA was categorized into one of three activity levels defined as light (<3.0 metabolic equivalents; LPA), moderate and vigorous (3.0 metabolic equivalents) activity. Partial correlation analysis was used to investigate the correlation between PA and cognition, with adjustments for age, sex and school years. We then carried out a multiple regression analysis to investigate the association of cognitive performance with PA, adjusting for insulin resistance or depressive mood. RESULTS: Partial correlation adjusted for age, sex and schooling years showed that LPA was significantly correlated with the Digit Symbol Substitution test, Trail Making Test (TMT) part A and TMT-B, whereas moderate and vigorous activity showed no correlations. Multiple regression analysis with several models with different adjustments showed that LPA was associated with the Digit Symbol Substitution test, TMT-A and TMT-B independently from insulin resistance or depressive mood. CONCLUSIONS: In the current study, we found that LPA was significantly associated with the performance of executive functional assessments. Geriatr Gerontol Int 2018; 18: 922-928.

MicroRNA-182 Regulates Neurite Outgrowth Involving the PTEN/AKT Pathway.

MicroRNAs are implicated in neuronal development and maturation. Neuronal maturation, including axon outgrowth and dendrite tree formation, is regulated by complex mechanisms and related to several neurodevelopmental disorders. We demonstrated that one neuron-enriched microRNA, microRNA-182 (miR-182), played a significant role in regulating neuronal axon outgrowth and dendrite tree formation. Overexpression of miR-182 promoted axon outgrowth and complexity of the dendrite tree while also increasing the expression of neurofilament-M and neurofilament-L, which provide structural support for neurite outgrowth. However, a reduction of miR-182 inhibited neurite outgrowth. Furthermore, we showed that miR-182 activated the AKT pathway by increasing AKT phosphorylation on S473 and T308 and inhibiting PTEN activity by increasing phosphorylation on S380. Inhibition of AKT activity with the PI3-K inhibitor LY294002 could downregulate AKT and PTEN phosphorylation and suppress axon outgrowth. In addition, we showed that BCAT2 might be the target of miR-182 that takes part in the regulation of neuronal maturation; blockage of endogenous BCAT2 promotes axon outgrowth and AKT activity. These observations indicate that miR-182 regulates axon outgrowth and dendrite maturation involving activation of the PTEN/AKT pathway.

Iron addiction: a novel therapeutic target in ovarian cancer.

Ovarian cancer is a lethal malignancy that has not seen a major therapeutic advance in over 30 years. We demonstrate that ovarian cancer exhibits a targetable alteration in iron metabolism. Ferroportin (FPN), the iron efflux pump, is decreased, and transferrin receptor (TFR1), the iron importer, is increased in tumor tissue from patients with high grade but not low grade serous ovarian cancer. A similar profile of decreased FPN and increased TFR1 is observed in a genetic model of ovarian cancer tumor-initiating cells (TICs). The net result of these changes is an accumulation of excess intracellular iron and an augmented dependence on iron for proliferation. A forced reduction in intracellular iron reduces the proliferation of ovarian cancer TICs in vitro, and inhibits both tumor growth and intraperitoneal dissemination of tumor cells in vivo. Mechanistic studies demonstrate that iron increases metastatic spread by facilitating invasion through expression of matrix metalloproteases and synthesis of interleukin 6 (IL-6). We show that the iron dependence of ovarian cancer TICs renders them exquisitely sensitive in vivo to agents that induce iron-dependent cell death (ferroptosis) as well as iron chelators, and thus creates a metabolic vulnerability that can be exploited therapeutically.

The Soluble VEGF Receptor sFlt-1 Contributes to Impaired Neovascularization in Aged Mice.

The mechanism by which angiogenesis declines with aging is not fully understood. Soluble vascular endothelial growth factor receptor 1 (VEGFR1) form (sFlt1) contributes to endothelial dysfunction in pathological conditions. However, the roles of sFlt1 in ischemia-induced neovascularizationof aged animals have not been investigated. To study aging-related sFlt1 change and its impact on ischemia-induced neovascularization, a hindlimb ischemia model was applied to young and aged mice. Blood flow imaging assay revealed that the blood flow recovery remained impaired throughout the follow-up period. At day 14, immunostaining showed lesser capillary formation in the aged mice. An ELISA showed that the aged mice had increased plasma sFlt-1 levels at indicated time points after surgery. On operative day 4, the aged ischemic muscles had decreased levels of p-VEGFR2 and p-Akt and increased levels of sFlt-1, Wnt5a, and SC35 genes or/and protein as well as increased numbers of inflammatory cells (macrophages and leucocytes) and matrix metalloproteinase-9 activity. Immnunofluorescence showed that Flt-1 was co-localized with CD11b+ macrophages of aged ischemic muscles. Hypoxia stimulated sFlt1 expression in CD11b+ cells of aged bone-marrow (BM), and this effect was diminished by siWnt5a. The cultured medium of aged mice BM-derived CD11b+ cells suppressed human endothelial cell (EC) and endothelial progenitor cell (EPC) angiogenic actions induced by VEGF, and these decreases were improved by treatment with siWnt5a-conditioned medium. Thus, aging appears to decline neovascularization in response to ischemic stress via the VEGFR2/Akt signaling inactivation in ECs and ECPs that is mediated by Wnt5a/SC35 axis activated macrophages-derived sFlt1 production in advanced age.

Activation of erbB2 and c-src in phorbol ester-treated mouse epidermis: possible role in mouse skin tumor promotion.

In recent work we showed that the EGF receptor (EGFr) was activated in tumor promoter treated mouse epidermis (Cell Growth & Differentiation, 6: 1447-1455, 1995). In the present study, we have investigated the possible role of other erbB family members in the process of tumor promotion. Both erbB2 and erbB3, but not erbB4, were expressed in cultured mouse keratinocytes and in mouse epidermis in vivo. In cultured mouse keratinocytes, EGF stimulated rapid tyrosine phosphorylation of erbB2 followed by a time-dependent degradation of erbB2 protein. Furthermore, an increase in erbB2:EGFr heterodimer formation was also induced by EGF. In contrast to the results with erbB2, EGF did not induce tyrosine phosphorylation, the degradation of erbB3, or erbB3:EGFr heterodimer formation in cultured keratinocytes. Further analyses revealed that c-src kinase activity was dramatically elevated in cultured mouse keratinocytes exposed to EGF. In mouse epidermis following multiple treatments with 12-O-tetradecanoylphorbol-13-acetate (TPA), the phosphotyrosine content of erbB2 was significantly elevated in a dose-dependent manner. Concomittantly, erbB2:EGFr heterodimer formation and c-src kinase activity were also elevated in TPA-treated epidermis. Structure-activity relationships with several phorbol ester analogs showed that the elevated phosphorylation of erbB2 in mouse epidermis followed closely with tumor promoting ability. Activation of erbB2 and c-src kinase were also observed in the epidermis of TGF alpha transgenic mice where expression of human TGF alpha was targeted to basal keratinocytes with the human K14 promoter. Collectively, the current data suggest that the activation of erbB2 in phorbol ester treated skin can be explained solely by a mechanism involving elevation of EGFr ligands and activation of the EGFr. In addition, activation of c-src may be an important downstream effector in mouse keratinocytes both in vivo and in vitro, following activation of the EGFr, erbB2, or both.

Differential localization and activity of the A- and B-forms of the human progesterone receptor using green fluorescent protein chimeras.

Subcellular localization and transcriptional activity of green fluorescent protein-progesterone receptor A and B chimeras (GFP-PRA and GFP-PRB) were examined in living mammalian cells. Both GFP-PRA and B chimeras were found to be similar in transcriptional activity compared with their non-GFP counterparts. GFP-PRA and PRA were both weakly active, while GFP-PRB and PRB gave a 20- to 40-fold induction using a reporter gene containing the full-length mouse mammary tumor virus long-terminal repeat linked to the luciferase gene (pLTRluc). Using fluorescence microscopy, nuclear/cytoplasmic distributions for the unliganded and hormone activated forms of GFP-PRA and GFP-PRB were characterized. The two forms of the receptor were found to have distinct intracellular distributions; GFP-PRA was found to be more nuclear than GFP-PRB in four cell lines examined. The causes for and implications of this differential localization of the A and B forms of the human PR are discussed.

Prevalence of glaucoma in the west of Ireland.

County Roscommon in the west of Ireland is a relatively remote rural area whose population of 55,000 is served by two community medical ophthalmologists and three optometrists. Eye surgical services are not available within the county. In order to assess the needs of the community for prevention of blindness from glaucoma, a simple random sample of the population of County Roscommon was taken for a community based glaucoma survey. A total of 2186 people over the age of 50 were examined which represented a 99.5% response rate. The high response rate was achieved by the community basis of the study and vigorous follow up of non-attenders. Intraocular pressure was measured using applanation tonometry, disc evaluation by both direct ophthalmoscopy and stereoscopic biomicroscopy, and visual field analysis using the Henson CFS 2000 and experimental computer controlled video perimetry. Diagnostic criteria were consistent with the preferred practice pattern of the American Academy of Ophthalmology. A crude prevalence of approximately 2% for primary open angle and normal tension glaucoma was found. The population profile of intraocular pressure showed a pattern which decreased with increasing age unlike the Framingham and Ferndale studies but similar to Japanese data.

Controlling cytoskeleton structure by phosphoinositide-protein interactions: phosphoinositide binding protein domains and effects of lipid packing.

Cell movement and resistance to mechanical forces are largely governed by the cytoskeleton, a three-dimensional network of protein filaments that form viscoelastic networks within the cytoplasm. The cytoskeleton underlying the plasma membrane of most cells is rich in actin filaments whose assembly and disassembly are regulated by actin binding proteins that are stimulated or inhibited by signals received and transmitted at the membrane/cytoplasm interface. Inositol phospholipids, or phosphoinositides, are potent regulators of many actin binding proteins, and changes in the phosphorylation of specific phosphoinositide species or in their spatial localization are associated with cytoskeletal remodeling in vitro. This review will focus on recent studies directed at defining the structural features of phosphoinositide binding sites in actin binding proteins and on the influence of the physical state of phosphoinositides on their ability to interact with their target proteins.