[Clinical characteristics and their influences on the survival of leptomeningeal metastasis derived from lung adenocarcinoma harboring epithelial growth factor receptor mutation].

Objective: To analyze the clinical characteristics of leptomeningeal metastasis (LM) patients from epithelial growth factor receptor (EGFR)-mutated lung adenocarcinoma, and their impacts on the survival of the patients. Methods: From July 2018 to July 2022, the clinicopathological data of 81 patients diagnosed as EGFR-mutated lung adenocarcinoma LM by cytopathology who admitted to the Department of Oncology of Xiangya Hospital of Central South University were retrospectively analyzed, including 33 males and 48 females. The age ranged from 31 to 76 years, with a median age of 54 years. All the 81 patients were followed up, with a median follow-up of 21.0 months (95%CI: 12.5 to 29.5 months). The Kaplan Meier method was used to draw survival curve. Cox proportional hazards regression model was used to analyze the impact of the factors on the survival of patients. Results: Among the 81 patients, the interval between the initial diagnosis of lung cancer and the pathological diagnosis of LM in cerebrospinal fluid (CSF) was 0-108 months, with a median interval of 14 months. Fifty-two patients (64.2%) used the third-generation epithelial growth factor receptor tyrosine kinase inhibitor (EGFR-TKIs), while 17 patients (21.0%) used EGFR-TKIs in combination with other drugs, and 12 patients (14.8%) were treated with best supportive care (BSC). Sixty patients (74.1%) had a Kanofsky performance status (KPS) score of less than 60 points, and 71 patients (87.7%) had brain parenchymal metastasis and/or spinal metastasis. Twenty-two patients (27.2%) used pemetrexed through intrathecal CSF, and 17 patients (21.0%) used pemetrexed through the Ommaya sac to the CSF of the ventricle. The incidence of adverse event related to the administration of pemetrexed through CSF was 64.1% (25/39), mainly manifested as myelosuppression, including 22 patients of leukocyte reduction, 25 patients of hemoglobin reduction, and 14 patients of platelet reduction. The median post-leptomeningeal metastasis overall survival (pLM-OS) in 81 patients was 11.0 (95%CI: 7.7-14.3) months. KPS score≥60 points (HR=0.407, 95%CI: 0.170-0.973, P=0.043), CSF cytology negative after treatment (vs persistent positive, HR=0.351, 95%CI: 0.155-0.792, P=0.012), intraventricular administration of pemetrexed (vs non intraventricular administration of pemetrexed, HR=0.319, 95%CI: 0.137-0.745, P=0.008) and the treatment with third-generation EGFR-TKIs after LM (vs EGFR-TKIs in combination with other drugs, HR=0.486, 95%CI: 0.237-0.998, P=0.049) were a factor affecting pLM-OS of patients. Conclusions: Brain parenchyma, or/and spine are the most sites where the LM patients concurrently metastasize. KPS score≥60 points and CSF cytology negative after treatment, intraventricular administration of pemetrexed and the treatment with third-generation EGFR-TKIs are indictors affecting pLM-OS of the patients.

[The clinical characteristics and treatment of children with disorders of sex development and early prevention of their siblings].

Disorders of sex development (DSD) is a class of diseases characterized by discordant phenotypes of sex chromosome karyotypes, gonads and external genitalia. The etiology is complex and the clinical manifestations are varied. Understanding the clinical characteristics of patients with various types of DSD help make accurate etiological diagnosis and prepare individualized treatment plans according to the etiology (including sex assignment, endocrine hormone replacement, surgery and fertility protection, etc.). Due to the increased risk of DSD in the second pregnancy of the parents of DSD patients, early preventive measures such as pre-pregnancy genetic counseling and prenatal diagnosis during pregnancy can effectively avoid or reduce the risk of DSD in their siblings.

[Viral pathogenic spectrum analysis of severe acute respiratory infection cases in Luohe City, Henan province from 2017 to 2019].

Objective: The purpose of this study was to investigate the characteristics of viral pathogen spectrum and the epidemiological characteristics of each viral pathogen in hospitalized cases associated with severe acute respiratory infection (SARI) in Luohe City, Henan Province from 2017 to 2019. Methods: Based the SARI Case Surveillance Platform, SARI cases were collected in Central Hospital of Luohe City, Henan Province from November 2017 to February 2019. In the end, 783 SARI cases were included, whose throat swabs were taken within 24 h of admission, as well as their demographic characteristics, onset time, clinical characteristics and other information recorded. At the same time, viral identification was performed, and the age and time distribution of each virus were analyzed. Results: The age of 783 SARI cases shown as M (P25, P75) was 3 (1, 5) years old, ranging from 1 month to 95 years old. Children under 5 years old were the majority (71.01%). The males (61.81%) were more than females (38.18%). Among the 783 SARI cases, a total of 9 kind of viruses were identified with 64.88% (508/783) of the throat swabs tested positive for at least one virus. The positive rate of influenza virus and human respiratory syncytial virus were both 20.18% (158 cases), which was the highest among all the detected respiratory virus. The co-infection rate was 15.84% (124/783), among which double infection was the most common, accounting for 85.48% (106/124) of the co-infected cases. And human respiratory syncytial virus, human rhinovirus and influenza virus were the most common pathogen in co-infection cases. Moreover, the viral positive rate was 68.71% in children aged 5 years and 63.27% in people aged 60-95 years. Influenza and human respiratory syncytial virus dominated in winter and spring, while human parainfluenza virus was the main infection in summer. Conclusion: Influenza virus and human respiratory syncytial virus were the main viruses in throat swabs of SARI cases from 2017 to 2019 in Luohe City, Henan Province. There were differences in the age and seasonal epidemiological characteristics of each virus.

The impact of isolated maternal hypothyroxinaemia on the incidence of large-for-gestational-age infants: the Ma'anshan Birth Cohort study.

OBJECTIVE: The purpose of this study was to investigate whether isolated maternal hypothyroxinaemia (IMH) is associated with risks of small/large-for-gestational-age (SGA/LGA) infants. DESIGN: Population-based prospective cohort study. SETTING: Ma'anshan Maternal and Child Health (MCH) clinics, China. POPULATION: Pregnant women with singleton births (n = 3178). METHODS: Descriptive statistics were calculated for the demographic characteristics of the mothers and their newborns. Linear regression was applied to estimate the association between thyroid hormone levels and birthweight. Logistic regression was performed to calculate the association between IMH and SGA/LGA. MAIN OUTCOME MEASURES: Outcomes included SGA/LGA. RESULTS: The prevalence of IMH, defined as a free thyroxine value (FT4) lower than the 2.5th percentile with normal thyroid stimulating hormone, was 2.5% (78/3080) and 2.5% (74/2999) in the first and second trimesters, respectively. Additionally, 306 (9.6%) and 524 (16.5%) infants were defined as SGA and LGA, respectively. No evidence supported the notion that IMH is associated with an increased risk for SGA in either the first [odds ratio (OR): 1.762, 95% confidence interval (CI): 0.759-4.089] or the second (OR: 0.763, 95% CI: 0.231-2.516) trimester. However, an increased risk of LGA was observed among IMH women in the second trimester (OR: 2.088, 95% CI: 1.193-3.654). Maternal TPO-Ab positivity in the second trimester increased the risk of SGA (OR: 2.094, 95% CI: 1.333-3.290). CONCLUSION: This study provides evidence that IMH is associated with LGA. FUNDING: This work was supported by the National Natural Science Foundation of China (No. 81330068). TWEETABLE ABSTRACT: Isolated maternal hypothyroxinaemia may increase the risk of large-for-gestational-age infants.

[Pregnancy intention and pregnancy-related anxiety in the second and third trimester: a birth cohort study].

Objective: To understand the association between pregnancy intention and pregnancy-related anxiety in the second and third trimester and its strength. Methods: A prospective cohort study was conducted in Ma'anshan, Anhui province. A total of 3 474 eligible pregnant women within 14 weeks of gestation were recruited. The information about their demographic characteristics were collected in early pregnancy. The completed questionnaire of pregnancy-related anxiety were asked to return in the second and third trimester. Logistic regression analysis was conducted to evaluate the association between pregnancy intention and pregnancy-related anxiety in the second and third trimester. Results: A total of 3 083 pregnant women were included in final analysis, The rate of unintentional pregnancy was 15.00% (n=461). The detection rates of pregnancy-related anxiety in the second and third trimester were 29.13% (n=898) and 30.36% (n=936). After controlling potential confounding factors, unintentional pregnancy increased the risk of pregnancy-related anxiety in the second trimester compared with intentional pregnancy (OR=1.85, 95%CI: 1.44-2.38); The risk of pregnancy-related anxiety also increased in the third trimester (OR=1.84, 95%CI: 1.44-2.35). Intentional pregnancy did not increase the risk of pregnancy-related anxiety in the second and third trimester. Conclusion: The study results suggests that unintentional pregnancy could increase the risk of pregnancy-related anxiety in the second and third trimester.

Efficient quantum circuits for dense circulant and circulant like operators.

Circulant matrices are an important family of operators, which have a wide range of applications in science and engineering-related fields. They are, in general, non-sparse and non-unitary. In this paper, we present efficient quantum circuits to implement circulant operators using fewer resources and with lower complexity than existing methods. Moreover, our quantum circuits can be readily extended to the implementation of Toeplitz, Hankel and block circulant matrices. Efficient quantum algorithms to implement the inverses and products of circulant operators are also provided, and an example application in solving the equation of motion for cyclic systems is discussed.

[Analysis on the performance evaluation of the Global Fund Malaria Programme in China from 2003 to 2013].

Objective: To analyze the performance of the 5 Global Fund Malaria Programmes in China from 2003 to 2013. Methods: All of the proposals, summaries, progress reports, survey reports, Monitoring& Evaluation reports, and performance rating reports of the 5 Global Fund Malaria Programmes in China and the epidemic data of program areas were collected for statistical analysis from 2012 to 2014. Symposiums were held with relevant experts from national and provincial Centers for Disease Control and Prevention, program managers and staffs from national and provincial Global Fund Malaria Programme offices. The completion of the relevant programme indicators (including the general grant information such as program areas, beneficiaries and funding; the implementation of malaria control measures; the performance of malaria control measures; the malaria incidence in the program areas; the prevalence of malaria parasites; and program management and performance evaluation) were analyzed, and the results: of the symposiums were summarized. Results The implementation period of the 5 Global Fund Malaria Programs were as follows: Round 1 from 2003 to 2008, Round 5 from 2006 to 2010, Round 6 from 2007 to 2012, Round 10 from 2012 to 2013, and National Strategy Application (NSA) from 2010 to 2012. Under the support of all the Global Fund Malaria Programs, a total of 11 936 726 fever cases received microscopic tests, 1 485 915 confirmed and suspected malaria cases were treated, 1 579 773 Long Lasting Insecticide-treated Nets were distributed, 3 414 633 regular nets were treated by insecticide, 40 298 284 primary and middle school students received health education on malaria control. Compared with the baseline value, the completion rates of each indicator increased after the implementation of the programs. The growth value ranged from 12.83% to 83.11%, among which the biggest growth was the value of the indicator'Percentage of households with at least one LLIN/ITN in target areas', and it increased from 9.2% (baseline value of 2006) to 92.31% (value of 2012). The malaria incidence in program areas has dropped significantly year by year, the annual reported malaria incidence in Yunnan and Hainan provinces decreased from 1 950/100 000, 3 850/100 000 in 2002 to 3.31/100 000, 0.15/100 000 in 2012, the P. falciparum malaria incidence in target counties in Hainan province decreased from 90.6/100 000 in 2002 to 0/100 000 in 2012. As from the implementation of NSA grant in 2010 to 2012, the annual reported malaria incidence in 92% of the 75 Type 1 counties was less than 1 per 10 000, 60.00% of Type 1 counties and 98.69% of the 687 Type 2 counties reported zero locally transmitted malaria cases. The Global Fund Secretariat had conducted a total of 37 performance evaluations, of which 9 have been rated as A1, 4 rated as A2, 19 rated as B1 and 5 rated as B1. Conclusions: The Global Fund Malaria Program in China has been closely integrated with the goal and task of National Malaria Control Program, reducing malaria burden in target areas, and pushing Chinese malaria control efforts to move from control to elimination.

The transmissibility estimation of influenza with early stage data of small-scale outbreaks in Changsha, China, 2005-2013.

Hundreds of small-scale influenza outbreaks in schools are reported in mainland China every year, leading to a heavy disease burden which seriously impacts the operation of affected schools. Knowing the transmissibility of each outbreak in the early stage has become a major concern for public health policy-makers and primary healthcare providers. In this study, we collected all the small-scale outbreaks in Changsha (a large city in south central China with ~7·04 million population) from January 2005 to December 2013. Four simple and popularly used models were employed to calculate the reproduction number (R) of these outbreaks. Given that the duration of a generation interval Tc = 2·7 and the standard deviation (s.d.) σ = 1·1, the mean R estimated by an epidemic model, normal distribution and delta distribution were 2·51 (s.d. = 0·73), 4·11 (s.d. = 2·20) and 5·88 (s.d. = 5·00), respectively. When Tc = 2·9 and σ = 1·4, the mean R estimated by the three models were 2·62 (s.d. = 0·78), 4·72 (s.d. = 2·82) and 6·86 (s.d. = 6·34), respectively. The mean R estimated by gamma distribution was 4·32 (s.d. = 2·47). We found that the values of R in small-scale outbreaks in schools were higher than in large-scale outbreaks in a neighbourhood, city or province. Normal distribution, delta distribution, and gamma distribution models seem to more easily overestimate the R of influenza outbreaks compared to the epidemic model.

[Previous medical or surgical abortions and subsequent risk of preterm birth: a birth cohort study].

Objective: To understand the association between medical abortion (MA) or surgical abortion (SA) and the risk of preterm birth (PTB) in subsequent pregnancy. Methods: The prospective cohort study was conducted in Ma'anshan, Anhui province. The information about demographic characteristics and previous MA or SA of 3 474 pregnant women were collected before 14 gestational weeks. Logistic regression analysis was conducted to compare the rates of preterm birth based on the history of previous MA or SA, and 3 256 live births were included in the analysis. Results: The PTB rate and spontaneous preterm birth (sPTB) rate were 4.12% (n=134) and 2.49% (n=81) respectively. Previous MA was associated with an increased risk of total PTB (RR=2.00, 95%CI: 1.04-3.85 for one MA and RR=3.58, 95%CI: 1.04-12.30 for two or more MAs) and sPTB (RR=2.51, 95% CI: 1.23-5.15). The risk of PTB in women with one SA (RR=0.67, 95%CI: 0.42-1.01) or more SA (RR=0.97, 95%CI: 0.51-1.85) did not differ significantly compared with the women with no history of SA. Conclusion: This study suggests that medical abortion could increase the risk of PTB or sPTB.

Effects of NELL2 on the regulation of GnRH expression and puberty in female rats.

Neural tissue-specific epidermal growth factor-like repeat domain-containing protein (NELL2) was previously found to play an important role in nerve growth, neural differentiation, neural elasticity, synaptic transport, and vesicle release. In this study, we examined the effect of NELL2 on gonadotropin-releasing hormone (GnRH) neurons and the initiation of puberty in female rats. We studied changes in NELL2 mRNA and protein expression at different stages of sexual development (postnatal days 30, 35, and 45) in female rats to determine the impact of NELL2 on GnRH mRNA expression. We also investigated the influence on the vulva-opening age by inhibiting NELL2 mRNA expression through lentiviral vector-mediated RNA interference. The intraventricular administration of an NELL2-interfering virus reduced NELL2 and GnRH expression at multiple stages of sexual development and delayed the age of vulva-opening in female rats. These results demonstrate that lentiviral-mediated RNA interference technology can be used for targeted regulation of sexual development in vivo. In addition, we found that NELL2 regulated the initiation of puberty in female rats.

Ferulic acid induces heme oxygenase-1 via activation of ERK and Nrf2.

This study investigated the effect of ferulic acid (FA) on the up-regulation of heme oxygenase-1 (HO-1) in lymphocytes and the molecular mechanisms involved. Lymphocytes were treated with FA (0.001-0.1 µM) for certain times. Cell viability, the activity and level of expression of HO-1, and signal pathways were analyzed. FA significantly upregulated HO-1 expression both at the level of mRNA and protein in lymphocytes. Moreover, FA induced NF-E2-related factor (Nrf2) nuclear translocation and transcriptional activity, which is upstream of FA induced HO-1 expression. In addition, lymphocytes treated with FA exhibited activation of extracellular regulated kinase (ERK) and treatments with U0126 (an ERK kinase inhibitor) attenuated the FA induced activation of Nrf2, resulting in a decrease in HO-1 expression. Zinc protoporphyrin (ZnPP, a HO-1 inhibitor) markedly suppressed cytoprotection from radiation-induced cell damage by FA. Results suggested that the ERK signaling pathway controlled the anti-oxidation of FA by regulating the expression of the antioxidant enzyme HO-1.

Differential detection of B virus and rhesus cytomegalovirus in rhesus macaques.

Non-human primate herpesviruses establish and maintain a lifelong persistent infection in immunocompetent hosts in the absence of clinical signs of disease. A fundamental issue for understanding the natural history of non-human primate herpesviruses is whether the viruses are maintained in a truly latent state or one characterized by a low level of chronic expression. To address this issue, a real-time PCR assay was developed to quantify Cercopithecine herpesvirus type 1 (B virus) DNA in mucosal fluids of rhesus macaques. This assay was rapid, sensitive (10 genome copies) and specific for B virus obtained from multiple species of macaques. The shedding profile of B virus was compared to another endemic herpesvirus, rhesus cytomegalovirus (RhCMV), in colony-reared monkeys. Mucosal swabs or saliva samples were taken daily from two groups of seropositive monkeys undergoing either a stressful relocation (group 1) or daily chair restraint (group 2). B virus DNA was detected in mucosal fluids from four animals relocated during the breeding season (group 1) but not from 10 animals moved at other times of the year. No B virus DNA was detected in any group 2 monkey. In contrast, RhCMV DNA was detected in the majority of animals of both groups 1 and 2. Detection of B virus DNA shedding is a relatively rare event associated with the breeding season, while RhCMV DNA is persistently detected in mucosal fluids of most monkeys.

Regulation of cardiac CFTR Cl(-) channel activity by a Mg(2+)-dependent protein phosphatase.

Dephosphorylation of the CFTR Cl(-) channel is known to be induced by both okadaic-acid- (OA-) sensitive and -insensitive protein phosphatases (PPs). In the present study, the effects of cytosolic free Mg(2+) on the cardiac CFTR Cl(-) current were examined in relation to the latter PP activity in guinea pig ventricular myocytes. Even when maintaining intracellular Mg-ATP at millimolar concentrations under whole-cell patch-clamp mode, cAMP-activated Cl(-) conductance was reversibly suppressed by cytosolic free Mg(2+), with an IC(50) of around 2.5 mmol/l. In contrast, changes in the cytosolic concentration of free Mg(2+) ([Mg(2+)](i)) had no effect on genistein-activated CFTR Cl(-) currents. The Mg(2+) effect on cAMP-activated CFTR Cl(-) conductance was completely reversed by application of anthracene-9-carboxylic acid (9-AC), which was previously shown to inhibit an OA-insensitive PP in cardiac myocytes. A 9-AC-sensitive fraction of endogenous PP activity in the extract of guinea pig ventricle was found to be activated by free Mg(2+) at millimolar concentrations but to be inactive at micromolar concentrations. The intracellular application of OA failed to activate basal Cl(-) conductance at millimolar [Mg(2+)](i). In the presence of OA, however, basal Cl(-) conductance became activated either by reducing [Mg(2+)](i) to micromolar concentrations or by applying 9-AC. Thus, we conclude that a Mg(2+)-dependent PP sensitive to 9-AC plays a key role in the cAMP-mediated regulation of cardiac CFTR Cl(-) channel at physiological [Mg(2+)](i)under both basal and cAMP-activated conditions. Also, it appears that the genistein-activated conformation of the cardiac CFTR channel is not sensitive to the Mg(2+)-dependent PP.

[Inhibitory effect of genistein on the proliferation of the anterior pituitary cells of rats].

The effects of tyrosine kinase inhibitor genistein on the proliferation of rat anterior pituitary cells and mouse AtT-20 cells were studied using techniques of cell culture, 3H-TdR incorporation, flow cytometric analysis and electron microscope. Genistein significantly inhibited the proliferation of rat anterior pituitary cells and mouse AtT-20 cells. Genistein (50 and 100 mumol/L) blocked the proliferation of AtT-20 cells at G0/G1 and G2/M phases and evoked an apoptotic peak of these cells with an apoptotic ratio of 19.9% and 36.4%. The apoptotic cells were also observed under the electron microscope. In consequence, genistein, as a tyrosine kinase inhibitor, can significantly inhibit the proliferation of pituitary cells possibly by inducing apoptosis, and the tyrosine kinase activity may play a key role in the proliferation and differentiation of pituitary cells.

Prenatal exposure to toluene results in abnormal neurogenesis and migration in rat somatosensory cortex.

Toluene inhalant abuse during pregnancy may result in growth-retarded microcephalic newborns who subsequently demonstrate developmental impairment. By using a rat model of toluene-abuse embryopathy, we studied the effects of prenatal toluene exposure on the generation and migration of cortical neurons. Dams were exposed by gavage to either corn oil or toluene diluted in corn oil on d 6-21 of gestation. The time of origin of cortical neurons was determined in the mature pups of dams injected with the thymidine analogue 5'-bromodeoxyuridine on 1 d during the period from d 13-21 of gestation. 5'-Bromodeoxyuridine-labeled neurons were identified by immunohistochemistry in a 400-microm-wide column of somatosensory cortex. The brains of the toluene-exposed pups had a significant reduction in the number of neurons within each cortical layer (p < 0.001). Depending on the cortical layer, the generation of neurons in the toluene-exposed pups was delayed by 1 or 2 d. In addition, the brains of the toluene-exposed pups also showed evidence of abnormal neuronal migration. However, there were no differences in either brain weight or body weight between the control and toluene-exposed pups. These observations suggest that although prenatal toluene exposure results in abnormal neuronal proliferation and migration, brain weight in the toluene-exposed pups may be preserved by enhanced development of glia or the neuropil.

Primate cytomegaloviruses encode and express an IL-10-like protein.

An open reading frame (ORF) with homology to interleukin-10 (IL-10) has been identified in rhesus cytomegalovirus (RhCMV). The IL-10-like protein is generated from a multispliced, polyadenylated early gene transcript encompassing part of the corresponding UL111A ORF of human CMV (HCMV). Immunological analyses confirm expression of the IL-10-like protein both in tissue culture and in RhCMV-infected rhesus macaques. Conserved ORFs were subsequently identified in human, baboon, and African green monkey CMV, and a fully processed transcript has been mapped in fibroblasts infected with the Towne strain of HCMV. The conservation of this previously unrecognized ORF suggests that the protein may play an essential role in primate CMV persistence and pathogenesis.

Pathogenesis of experimental rhesus cytomegalovirus infection.

Human cytomegalovirus (HCMV) establishes and maintains a lifelong persistence following infection in an immunocompetent host. The determinants of a stable virus-host relationship are poorly defined. A nonhuman primate model for HCMV was used to investigate virological and host parameters of infection in a healthy host. Juvenile rhesus macaques (Macaca mulatta) were inoculated with rhesus cytomegalovirus (RhCMV), either orally or intravenously (i.v. ), and longitudinally necropsied. None of the animals displayed clinical signs of disease, although hematologic abnormalities were observed intermittently in i.v. inoculated animals. RhCMV DNA was detected transiently in the plasma of all animals at 1 to 2 weeks postinfection (wpi) and in multiple tissues beginning at 2 to 4 wpi. Splenic tissue was the only organ positive for RhCMV DNA in all animals. The location of splenic cells expressing RhCMV immediate-early protein 1 (IE1) in i.v. inoculated animals changed following inoculation. At 4 to 5 wpi, most IE1-positive cells were perifollicular, and at 25 wpi, the majority were located within the red pulp. All animals developed anti-RhCMV immunoglobulin M (IgM) antibodies within 1 to 2 wpi and IgG antibodies within 2 to 4 wpi against a limited number of viral proteins. Host reactivity to RhCMV proteins increased in titer (total and neutralizing) and avidity with time. These results demonstrate that while antiviral immune responses were able to protect from disease, they were insufficient to eliminate reservoirs of persistent viral gene expression.

Regulation of ROMK1 channel by protein kinase A via a phosphatidylinositol 4,5-bisphosphate-dependent mechanism.

ROMK inward-rectifier K+ channels control renal K+ secretion. The activity of ROMK is regulated by protein kinase A (PKA), but the molecular mechanism for regulation is unknown. Having found that direct interaction with membrane phosphatidylinositol 4, 5-bisphosphate (PIP2) is essential for channel activation, we investigate here the role of PIP2 in regulation of ROMK1 by PKA. By using adenosine-5'-[gamma-thio]triphosphate) (ATP[gammaS]) as the substrate, we found that PKA does not directly activate ROMK1 channels in membranes that are devoid of PIP2. Rather, phosphorylation by PKA + ATP[gammaS] lowers the concentration of PIP2 necessary for activation of the channels. In solution-binding assays, anti-PIP2 antibodies bind PIP2 and prevent PIP2-channel interaction. In inside-out membrane patches, antibodies inhibit the activity of the channels. PKA treatment then decreases the sensitivity of ROMK1 for inhibition by the antibodies, indicating an enhanced interaction between PIP2 and the phosphorylated channels. Conversely, mutation of the PKA phosphorylation sites in ROMK1 decreases PIP2 interaction with the channels. Thus, PKA activates ROMK1 channels by enhancing PIP2-channel interaction.

[Effects of monocarboxylic acid derivatives on cardiac ventricular CFTR Cl- channels in guinea pig].

Using the whole-cell recording technique, the effects of monocarboxylic acid derivatives on cystic fibrosis transmembrane conductance regulator (CFTR) Cl(-)-channel were examined in guinea pig ventricular myocytes. Anthracene-9-carboxylic acid (9-AC) added to the bath solution further enhanced the outward component of isoproterenol-induced currents in a reversible manner, whereas 5-nitro-2-(3-phenylpropylamino) benzoic acid (NPPB) or diphenylamine-2-carboxylic acid (DPC) induced a biphasic effect on the currents. Either NPPB or DPC first produced a transient increase in the outward component of current before ensuing inhibition. Intracellular NPPB was found to potentiate isoproterenol-activated currents. It is concluded that these monocarboxylic acid derivatives have different binding sites in cardiac ventricular myocytes, which might partially account for the varied effects in blocking anion channels.