4-hydroxynonenal (4-HNE) mediates many pathological effects of oxidative and electrophilic stress and signals to activate cytoprotective gene expression regulated by NF-E2-related factor 2 (Nrf2). By exhibiting very high levels of 4-HNE-conjugating activity, the murine glutathione transferase alpha 4 (GSTA4-4) helps regulate cellular 4-HNE levels. To examine the role of 4-HNE in vivo, we disrupted the murine Gsta4 gene. Gsta4-null mice exhibited no cardiac phenotype under normal conditions and no difference in cardiac 4-HNE level as compared to wild-type (WT) mice. We hypothesized that the Nrf2 pathway might contribute an important compensatory mechanism to remove excess cardiac 4-HNE in Gsta4-null mice. Cardiac nuclear extracts from Gsta4-null mice exhibited significantly higher Nrf2 binding to antioxidant-response elements (AREs). We also observed responses in critical Nrf2 target gene products: elevated Sod2, Cat, and Akr1b7 mRNA levels and significant increases in both cardiac anti-oxidant and anti-electrophile enzyme activities. Gsta4-null mice were less sensitive and maintained normal cardiac function following chronic doxorubicin (DOX) treatment, known to increase cardiac 4-HNE levels. Hence, in the absence of GSTA4-4 to modulate both physiological and pathological 4-HNE levels, the adaptive Nrf2 pathway may be primed to contribute to a preconditioned cardiac phenotype in the Gsta4-null mouse.

A valid sham control is important for determining the efficacy and effectiveness of repetitive transcranial magnetic stimulation (rTMS) as an experimental and clinical tool. Given the manner in which rTMS is applied, separately or in combination with self-regulatory approaches, and its intended impact on brain states, a valid sham control of this type may well serve as a meaningful control for biofeedback studies, where efforts to develop a credible control have often been less than ideal. This study examined the effectiveness of focal electrical stimulation of the frontalis muscle as a sham technique for blinding participants to high-frequency rTMS over the dorso-lateral prefrontal cortex (DLPFC) at durations, intensities, and schedules of stimulation similar to many clinical applications. In this within-subjects single blind design, 19 participants made guesses immediately after receiving 54 counterbalanced rTMS sessions (sham, 10Hz, 20Hz); 7 (13%) of the guesses were made for sham, 31 (57%) were made for 10Hz, and 16 (30%) were made for 20Hz. Participants correctly guessed the sham condition 6% (CI: 1%, 32%) of the time, which is less than the odds of chance (i.e., of guessing at random, 33%); correctly guessed the 10Hz condition 66% (CI: 43%, 84%) of the time, which was greater than chance; and correctly guessed the 20Hz condition 41% (CI: 21%, 65%) of the time, which was no different than chance. Focal electrical stimulation therefore can be an effective sham control for high-frequency rTMS of the DLPFC, as well as for active biofeedback interventions. Participants were unaware that electrical stimulation was, in fact, sham rTMS.

Purpose: Amantadine is commonly used to treat Parkinson’s disease. A case of myoclonus and asterixis was associated with amantadine is reported. Case Summary: An 80-year-old man with Parkinson’s disease diagnosed in 2015 was started on amantadine for treatment of progressive tremor and orofacial dyskinesias induced by levodopa. He took amantadine 100mg orally daily for 7 days, then increased to 100mg twice a day thereafter. The patient complained of “worsening tremor” after 9 days and amantadine was decreased to 100mg daily. After 1 month on this dose, the patient reported that his “tremor” persisted and experienced visual hallucinations. His examination demonstrated diffuse myoclonus throughout his extremities and trunk, as well as asterixis when attempting to stand or holding his arms antigravity. Laboratory testing for renal and hepatic failure was unrevealing. Amantadine was reduced to 50mg daily for 4 days and then discontinued. Myoclonus resolved 3 days after discontinuation of amantadine. Conclusion: While amantadine-induced myoclonus is rare, health care providers should be vigilant in monitoring for signs and symptoms of myoclonus following amantadine initiation.

As the fat body is a critical tissue for mosquito development, metamorphosis, immune and reproductive system function, characterization of regulatory modules targeting gene expression to the female mosquito fat body at distinct life stages is much needed for multiple, varied strategies for controlling vector-borne diseases such as dengue and malaria. The hexameric storage protein, Hexamerin-1.2, of the mosquito, Aedes atropalpus, is female-specific and uniquely expressed in the fat body of fourth-instar larvae and young adults. We have identified in the Hex-1.2 gene, a short regulatory module that directs female-, tissue-, and stage-specific lacZ reporter gene expression using a heterologous promoter in transgenic lines of the dengue vector, Aedes aegypti. Male transgenic larvae and pupae of one line expressed no E. coli β-galactosidase or transgene product; in two other lines reporter gene activity was highly female-biased. All transgenic lines expressed the reporter only in the fat body. However, lacZ mRNA levels were no different in males and females at all stages examined, suggesting that the gene regulatory module drives female-specific expression by post-transcriptional regulation in the heterologous mosquito. This regulatory element from the Hex-1.2 gene thus provides a new molecular tool for transgenic mosquito control as well as functional genetic analysis in aedine mosquitoes.

4‐hydroxynonenal (4‐HNE) mediates both pathological effects of oxidative stress and signaling from mitochondrion to nucleus to elicit transcriptional responses that are largely controlled by Nrf2. As elevated levels of 4‐HNE are associated with several cardiovascular diseases, metabolism of 4‐HNE is critical. The murine glutathione transferase A4–4 (GSTA4–4) exhibits very high levels of 4‐HNE‐conjugating activity, which is reduced in the hearts of Gsta4 knockout (KO) mice. KO mice exhibited no cardiac pathology; cardiac 4‐HNE levels were no different in WT and KO mice. We examined how Gsta4 KO mice react to high cardiac 4‐HNE accumulation following chronic administration of doxorubicin (DOX) (5 mg/kg, 4 weekly injections). Cardiac function was assessed by echocardiography. While untreated mice exhibited similar basal cardiac function, WT mice were more sensitive to DOX, resulting in a significantly higher mortality rate. Ejection Fraction (EF) fell in both groups after the first DOX injection. At day 21, EF was significantly lower in WT mice as compared to baseline whereas EF in KO mice recovered to the basal value. Changes in fractional shortening correlated with EF in both groups. We concluded that Gsta4 KO mice exhibited resistance to DOX cardiomyopathy and will explore how Nrf2‐induction of SOD and catalase may contribute to a cardioprotective effect. Supported by a College of Medicine Pilot Study Grant.