A clinical drug library screen identifies clobetasol propionate as an NRF2 inhibitor with potential therapeutic efficacy in KEAP1 mutant lung cancer.

The Kelch-like ECH-associated protein 1 (KEAP1)-nuclear factor E2-related factor 2 (NRF2)pathway has a central role in cellular antioxidant defense. NRF2 activation due to KEAP1 or NRF2 mutations occurs frequently in many cancers, suggesting that NRF2 inhibition could be a promising therapeutic strategy. However, no potent NRF2 inhibitors are clinically available to date. To develop potent NRF2 inhibitors for therapeutic purpose, we screened ~4000 clinical compounds and determined clobetasol propionate (CP) as the most potent NRF2 inhibitor. Mechanistically, CP prevented nuclear accumulation and promoted ß-TrCP-dependent degradation of NRF2 in a glucocorticoid receptor- and a glycogen synthase kinase 3 (GSK3)-dependent manner. As a result, CP induced oxidative stress and strongly suppressed the anchorage-independent growth of tumors with KEAP1 mutation, but not with the wild-type KEAP1. Further, CP alone or in combination with rapamycin strongly inhibited the in vitro and in vivo growth of tumors harboring mutations in KEAP1 or both KEAP1 and LKB1 that are frequently observed in lung cancer. Thus, CP could be a repurposed therapeutic agent for cancers with high NRF2 activity. We also proposed that the use CP and rapamycin in combination could be a potential therapeutic strategy for tumors harboring both KEAP1 and LKB1 mutations.

Altered expression of E-cadherin and β-catenin in malignant transformation of sinonasal inverted papillomas.

BACKGROUND: E-cadherin and β-catenin are molecules that mediate cell-cell adhesion in normal epithelium. Aberrant expression of these adhesion molecules results in the loss of intercellular adhesion, with possible cell transformation and tumour progression. We determined the role of E-cadherin and β-catenin in the pathogenesis of sinonasal inverted papilloma (IP) and its malignant transformation. METHODS: We determined the expression of E-cadherin and β-catenin by immunohistochemistry in paraffin-embedded tissue of 21 subjects with nasal polyps, 56 with IPs, 7 IPs with dysplasia and 18 IPs with squamous cell carcinoma (SCC). The clinicopathological variables of the IPs with SCC correlated with the degree of expression of E-cadherin and β-catenin. RESULTS: The degree of expression of E-cadherin and β-catenin in the cell membrane was significantly lower in IPs with SCC than in nasal polyps and IPs. The degree of expression of β-catenin was significantly lower in IPs with SCC with a malignant proportion > 50% compared to a malignant proportion ≤ 50%. However, there was no significant association between the degree of expression of E-cadherin and β-catenin and clinicopathological variables, such as age, gender, T stage, tumour differentiation, or SCC type (metachronous vs. synchronous). In addition, there was no significant relationship between recurrence or survival rate in IPs with SCC and the degree of expression of E-cadherin or β-catenin in the cell membrane or nuclear β-catenin. CONCLUSION: Decreased expression of E-cadherin and β-catenin in the cell membrane may be associated with carcinogenesis of IPs and help predict malignant transformation in sinonasal IPs.

Tunability of mobility and conductivity over large ranges in poly(3,3'''-didodecylquaterthiophene)/insulating polymer composites.

Semiconducting polymers are currently being considered as active layers in field-effect transistors, in which high charge carrier mobility and low off conductivity are important. For other applications, such as certain spintronic mechanisms, the opposite characteristics are desirable. Blending such polymers with insulating polymers would be expected to lower the mobility. In this paper, we report that the use of hydrocarbon polymers such as polystyrene as insulators generally raises the mobility when the semiconducting polymer is poly(bisdodecylquaterthiophene). A high mobility value of nearly 0.1 cm(2)/V.s was obtained for an optimal blend. While this is counterintuitive, it is consistent with a few other recent reports. In order to lower the mobility significantly, a much more polar and irregular blending agent is needed. The further addition of tetrafluorotetracyanoquinodimethane as a dopant gave a rare low mobility/high conductivity combination of properties, with a charge carrier density on the order of 10(19) cm(-3). Thus, mobility and conductivity were tuned somewhat independently over 3 and 4 orders of magnitude, respectively.

Lack of allosteric modulation of striatal GABA(A) receptor binding and function after cocaine sensitization.

GABA(A) receptor binding after repeated cocaine has been shown to be either increased as indicated by benzodiazepine binding or decreased as indicated by convulsant-site binding. We measured the GABA binding site with [3H]-muscimol binding to GABA(A) receptors and found no differences between saline- and cocaine-sensitized rats. Allosteric modulation of [3H]-muscimol binding with flunitrazepam was also unchanged after cocaine sensitization. In addition, [3H]-flunitrazepam binding and allosteric modulation of [3H]-flunitrazepam binding with GABA was unchanged after 1 day withdrawal from repeated cocaine. GABA(A) receptor function and allosteric modulation of GABA(A) receptor function measured by GABA-stimulated Cl(-) uptake was also unchanged after withdrawal from repeated cocaine. Finally, in vitro cocaine reduced GABA(A) receptor function in striatal microsacs of saline- and cocaine-treated rats. In conclusion, repeated cocaine did not change the coupling of the GABA(A) receptor between the GABA and benzodiazepine (BZD) binding site after 1 day withdrawal.

A study on psychological strain in IVF patients.

PURPOSE: The objectives of this study were to compare average stress levels in infertile women to fertile women, to determine the stress levels whether the patients was pregnant or not pregnant, and to examine for a cross-section of infertile patients in different stages of medical investigation for the infertility. METHODS: One hundred thirty-eight women receiving medical treatment for infertility attended the program. The State Trait Anxiety Inventory (STAI) and the Beck Depression Inventory (BDI) of perceived stress associated with the infertility was the outcome measure. RESULTS: Infertile women showed significant increases in trait anxiety and depressive symptoms than the fertile women. Anxiety and depression in the in vitro fertilization (IVF)-failed women were significantly higher than the IVF-success women. According to the duration of infertility, STAI and BDI were moderately elevated in the first stage (< 3 year). There was a trend of a decreasing psychological stress with an advanced infertility duration. On depression scales, the intermediate and final duration of infertility patients showed less symptomatology than the first-stage patients. Contrary to the expectation, demographic factors such as religion and husband cooperation were not related to the experience of stress. CONCLUSIONS: We must pay an attention to the infertile patient, especially from the initial infertility workup. We recommend psychological counselling for IVF-failed patients.

Influence of female age on pregnancy outcome in in vitro fertilization and embryo transfer patients undergoing intracytoplasmic sperm injection.

PURPOSE: To determine the influence of female age on the outcomes of ICSI in IVF-ET patients. METHODS: One hundred and seventy-five couples underwent 352 cycles of ICSI. The quality of oocytes and embryos, fertilization rate, and pregnancy outcomes were retrospectively evaluated according to female age; < 30 years in Group A (49 cycles), 30-34 in Group B (177 cycles), 35-39 in Group C (97 cycles), and > or = 40 in Group D (29 cycles). RESULTS: The fertilization rates were not significantly different among the age groups. Significant negative linear correlations were observed between female age and the numbers of oocytes retrieved and embryos transferred, and cumulative embryo score. Clinical pregnancy rates were significantly decreased and spontaneous abortion rate increased with advancing age. CONCLUSIONS: Female age may be a prognostic indicator in ICSI program.

Increased acute cocaine sensitivity and decreased cocaine sensitization in GABA(A) receptor beta3 subunit knockout mice.

The role of the GABA(A) receptor beta3 subunit in determining acute cocaine sensitivity and behavioral sensitization to repeated cocaine was measured in mice missing both (-/-), one (+/-), or neither (+/+) allele of the beta3 gene. Locomotor stimulation induced by one cocaine injection (20 mg/kg, i.p.) was found to be greater in -/- mice compared with +/+ mice, whereas cocaine-induced behaviors were intermediate in +/- mice. Amphetamine did not cause greater locomotor responses in -/- mice, suggesting that the increased sensitivity of -/- mice to cocaine does not generalize to other psychomotor stimulants. GABA-stimulated chloride uptake was 51% lower in striatum of -/- mice compared with +/+ mice, but only 27% lower in cortex. After 14 daily cocaine injections, the behavioral response to cocaine was increased in +/+ and +/- mice, but was not increased further in -/- mice. Additionally, repeated cocaine exposure decreased striatal GABA(A) receptor function in +/+ and +/- mice. In -/- mice, GABA(A) receptor function was not decreased any further by repeated cocaine injections. Thus, alterations in the beta3 subunit may be responsible for determining the behavioral responses induced by acute and repeated cocaine treatment, as well as mediating the neurochemical adaptation that occurs during sensitization to repeated cocaine.

The outcome of sperm retrieval and intracytoplasmic sperm injection in patients with obstructive azoospermia: impact of previous tuberculous epididymitis.

PURPOSE: Our purpose was to investigate the influence of previous tuberculous epididymitis in patients with obstructive azoospermia on the outcome of sperm retrieval and intracytoplasmic sperm injection (ICSI). METHODS: Eighty-eight cycles of ICSI were performed in 44 patients with obstructive azoospermia; 16 cycles (7 patients) with tuberculous obstructive azoospermia and 72 cycles (37 patients) with nontuberculous obstructive azoospermia. RESULTS: The rates of fertilization and embryo cleavage were comparable, and there was no significant difference in the clinical pregnancy rate per fresh transfer between the two groups. The rates of embryo implantation and clinical miscarriage were also comparable. CONCLUSIONS: Embryo quality and pregnancy outcome in sperm retrieval and ICSI were comparable in both the tuberculous and the nontuberculous obstructive azoospermia patients. Although there was a preponderance of testicular sperm used in the tuberculous obstructive azoospermia group, our results suggest that previous tuberculous epididymitis in patients with obstructive azoospermia does not affect the outcome of sperm retrieval and ICSI.

Endogenous GABA release is reduced in the striatum of cocaine-sensitized rats.

The magnitude of behavioral sensitization to cocaine is correlated with decreased striatal GABA(A) receptor function. We examined whether GABA release from striatal slices is also altered in cocaine-treated rats. Behavioral sensitization was measured in rats receiving either saline or cocaine (15 mg kg(-1)) daily for 14 days. Cocaine-treated rats showed a significant increase in locomotion and stereotypy over days. Potassium-stimulated endogenous GABA release was measured from superfused striatal slices of these rats. GABA release was significantly decreased in cocaine-treated rats. However, striatal slices preloaded with [(3)H]GABA exhibited a slight but significant increase in release after cocaine sensitization. Similar treatment with a nonsensitizing dose of cocaine (7.5 mg kg(-1)) did not change endogenous GABA release. Saline- and cocaine-treated rats showed no differences in striatal glutamic acid decarboxylase activity at either a saturating or K(m) concentration of glutamate. Therefore, the decrease in endogenous GABA release is not due to a decrease in GABA synthesis, but may reflect changes in GABA storage pools. These data are consistent with an overall decrease in GABA transmission, both pre- and postsynaptically, in the striatum of sensitized rats, which could contribute to enhanced striatal output and behavioral sensitization.

Antisense inhibition of striatal GABAA receptor proteins decreases GABA-stimulated chloride uptake and increases cocaine sensitivity in rats.

The functional status of striatal GABAA receptors appears to be inversely related to the magnitude of cocaine-induced behaviors. Exposure of striatum to antisense oligodeoxynucleotides (ASODNs) targeted to the mRNAs for the alpha 2 and the beta 3 subunits of the GABAA receptor should decrease expression of receptor proteins and therefore might be expected to increase cocaine sensitivity. ASODNs, scrambled ODNs or saline were injected into right lateral ventricle of rats and behavioral responses to cocaine were tested 18-20 h after treatment. Animals injected separately with alpha 2 or beta 3 ASODNs exhibited increased behavioral sensitivity to cocaine compared to rats injected with saline or scrambled ODNs including performing more 360 degrees turns to the left than to the right. There was significantly less GABA-stimulated Cl uptake in right striatum compared to left striatum of ASODN-treated rats with no significant difference between sides in control animals. Specific binding to benzodiazepine and convulsant sites on the GABAA receptor was not selectively altered by ASODN treatment. Combined alpha 2 beta 3 ASODN treatment did not affect either cocaine sensitivity or GABAA receptor function. There was no difference between the density of Nissl stained cells in the left and right edges of striatum in control or ASODN-treated rats indicating the absence of significant neurotoxic effects of the ASODN treatment. Injection of fluorescein-conjugated ASODNs indicated that ASODN is present in striatum at times during which behavioral and neurochemical indices of GABA receptor function are decreased. Thus, the functional status of GABAA receptors in striatum may be involved in determining cocaine sensitivity.

Simultaneous cocaine exposure abolishes ethanol tolerance.

We measured changes in locomotor impairment in rats caused by ethanol exposure either given alone or simultaneously with cocaine. An initial ethanol injection (2.1 g/kg, intraperitoneally (i.p.) disrupted rotorod performance and this disruption was not significantly affected by the cocaine injection (15 mg/kg, i.p.). After 13 daily drug treatments, performance in the ethanol group was significantly improved whereas in the cocaine+ethanol group, performance remained disrupted to the same extent throughout testing (49 +/- 14 min). Cocaine sensitization developed after repeated exposure and this sensitization was greater in the cocaine+ethanol group. Next, all groups were tested with simultaneous ethanol and cocaine. Tolerance was not diminished in the ethanol group, whereas groups receiving saline, cocaine, or cocaine+ethanol exhibited equally disrupted behavior. During an ethanol-only test, the cocaine+ethanol group also did not respond differently from groups receiving saline or cocaine alone. There was no difference in tolerance of the GABAA receptor to ethanol enhancement in cortical microsacs from the ethanol and cocaine+ethanol groups, nor did cocaine affect blood ethanol levels after initial or repeated exposure. A non-sensitizing dose of cocaine (7.5 mg/kg, i.p.) had no effect on the development or expression of ethanol tolerance. Cocaine disruption of ethanol tolerance thus appears to be partly due to interference of expression of ethanol tolerance by cocaine sensitization and partly due to inhibition of the development of ethanol tolerance by non-GABAergic mechanisms.