Cholesterol inhibits human voltage-gated proton channel hHv1.

Although human sperm is morphologically mature in the epididymis, it cannot fertilize eggs before capacitation. Cholesterol efflux from the sperm plasma membrane is a key molecular event essential for cytoplasmic alkalinization and hyperactivation, but the underlying mechanism remains unclear. The human voltage-gated proton (hHv1) channel functions as an acid extruder to regulate intracellular pHs of many cell types, including sperm. Aside from voltage and pH, Hv channels are also regulated by distinct ligands, such as Zn2+ and albumin. In the present work, we identified cholesterol as an inhibitory ligand of the hHv1 channel and further investigated the underlying mechanism using the single-molecule fluorescence resonance energy transfer (smFRET) approach. Our results indicated that cholesterol inhibits the hHv1 channel by stabilizing the voltage-sensing S4 segment at resting conformations, a similar mechanism also utilized by Zn2+. Our results suggested that the S4 segment is the central gating machinery in the hHv1 channel, on which voltage and distinct ligands are converged to regulate channel function. Identification of membrane cholesterol as an inhibitory ligand provides a mechanism by which the hHv1 channel regulates fertilization by linking the cholesterol efflux with cytoplasmic alkalinization, a change that triggers calcium influx through the CatSper channel. These events finally lead to hyperactivation, a remarkable change in the mobility pattern indicating fertilization competence of human sperm.

Voltage sensor dynamics of a bacterial voltage-gated sodium channel NavAb reveal three conformational states.

High-resolution structures of voltage-gated sodium channels (Nav) were first obtained from a prokaryotic ortholog NavAb, which provided important mechanistic insights into Na+ selectivity and voltage gating. Unlike eukaryotic Navs, the NavAb channel is formed by four identical subunits, but its ion selectivity and pharmacological profiles are very similar to eukaryotic Navs. Recently, the structures of the NavAb voltage sensor at resting and activated states were obtained by cryo-EM, but its intermediate states and transition dynamics remain unclear. In the present work, we used liposome flux assays to show that purified NavAb proteins were functional to conduct both H+ and Na+ and were blocked by the local anesthetic lidocaine. Additionally, we examined the real-time conformational dynamics of the NavAb voltage sensor using single-molecule FRET. Our single-molecule FRET measurements on the tandem NavAb channel labeled with Cy3/5 FRET fluorophore pair revealed spontaneous transitions of the NavAb S4 segment among three conformational states, which fitted well with the kinetic model developed for the S4 segment of the human voltage-gated proton channel hHv1. Interestingly, even under strong activating voltage, the NavAb S4 segment seems to adopt a conformational distribution similar to that of the hHv1 S4 segment at a deep resting state. The conformational behaviors of the NavAb voltage sensor under different voltages need to be further examined to understand the mechanisms of voltage sensing and gating in the canonical voltage-gated ion channel superfamily.

Demographic and Clinical Characteristics of Mpox in Persons Who Had Previously Received 1 Dose of JYNNEOS Vaccine and in Unvaccinated Persons - 29 U.S. Jurisdictions, May 22-September 3, 2022.

As of November 14, 2022, monkeypox (mpox) cases had been reported from more than 110 countries, including 29,133 cases in the United States.* Among U.S. cases to date, 95% have occurred among males (1). After the first confirmed U.S. mpox case on May 17, 2022, limited supplies of JYNNEOS vaccine (Modified Vaccinia Ankara vaccine, Bavarian Nordic) were made available to jurisdictions for persons exposed to mpox. JYNNEOS vaccine was approved by the Food and Drug Administration (FDA) in 2019 as a 2-dose series (0.5 mL per dose, administered subcutaneously) to prevent smallpox and mpox disease.† On August 9, 2022, FDA issued an emergency use authorization to allow administration of JYNNEOS vaccine by intradermal injection (0.1 mL per dose) (2). A previous report on U.S. mpox cases during July 31-September 3, 2022, suggested that 1 dose of vaccine offers some protection against mpox (3). This report describes demographic and clinical characteristics of cases occurring ≥14 days after receipt of 1 dose of JYNNEOS vaccine and compares them with characteristics of cases among unvaccinated persons with mpox and with the vaccine-eligible vaccinated population in participating jurisdictions. During May 22-September 3, 2022, among 14,504 mpox cases reported from 29 participating U.S. jurisdictions,§ 6,605 (45.5%) had available vaccination information and were included in the analysis. Among included cases, 276 (4.2%) were among persons who had received 1 dose of vaccine ≥14 days before illness onset. Mpox cases that occurred in these vaccinated persons were associated with lower percentage of hospitalization (2.1% versus 7.5%), fever, headache, malaise, myalgia, and chills, compared with cases in unvaccinated persons. Although 1 dose of JYNNEOS vaccine offers some protection from disease, mpox infection can occur after receipt of 1 dose, and the duration of protection conferred by 1 dose is unknown. Providers and public health officials should therefore encourage persons at risk for acquiring mpox to complete the 2-dose vaccination series and provide guidance and education regarding nonvaccine-related prevention strategies (4).

Effects of bridge construction on songbirds and small mammals at Blennerhassett Island, Ohio River, USA.

Construction of man-made objects such as roads and bridges may have impacts on wildlife depending on species or location. We investigated songbirds and small mammals along the Ohio River, WV, USA at a new bridge both before and after construction and at a bridge crossing that was present throughout the study. Comparisons were made at each site over three time periods (1985-1987 [Phase I] and 1998-2000 [Phase II] [pre-construction], 2007-2009 [Phase III] [post-construction]) and at three distances (0, 100, 300 m) from the bridge or proposed bridge location. Overall, 70 songbirds and 10 small mammals were detected during the study. Cliff swallows (Petrochelidon pyrrhonota) and rock pigeons (Columba livia) showed high affinity for bridges (P < 0.05). Combined small mammal abundances increased between Phases I and II (P < 0.05), but did not differ between Phases II and III (P > 0.05). Species richness and diversity for songbirds and small mammals did not differ before and after bridge construction (P > 0.05). We found that most species sampled did not respond to the bridge crossing, and believe that the bridge is not causing any measurable negative density impacts to the species we investigated. The new bridge does provide habitat for exotic rock pigeons that are adjusted to man-made structures for nesting.

Structural dynamics determine voltage and pH gating in human voltage-gated proton channel.

Voltage-gated proton (Hv) channels are standalone voltage sensors without separate ion conductive pores. They are gated by both voltage and transmembrane proton gradient (i.e., ∆pH), serving as acid extruders in most cells. Like the canonical voltage sensors, Hv channels are a bundle of four helices (named S1 -S4), with the S4 segment carrying three positively charged Arg residues. Extensive structural and electrophysiological studies on voltage-gated ion channels, in general, agree on an outwards movement of the S4 segment upon activating voltage, but the real-time conformational transitions are still unattainable. With purified human voltage-gated proton (hHv1) channels reconstituted in liposomes, we have examined its conformational dynamics, including the S4 segment at different voltage and pHs using single-molecule fluorescence resonance energy transfer (smFRET). Here, we provide the first glimpse of real-time conformational trajectories of the hHv1 voltage sensor and show that both voltage and pH gradient shift the conformational dynamics of the S4 segment to control channel gating. Our results indicate that the S4 segment transits among three major conformational states and only the transitions between the inward and outward conformations are highly dependent on voltage and pH. Altogether, we propose a kinetic model that explains the mechanisms underlying voltage and pH gating in Hv channels, which may also serve as a general framework for understanding the voltage sensing and gating in other voltage-gated ion channels.

Neuromuscular electrical stimulation pulse duration and maximum tolerated muscle torque.

Neuromuscular electrical stimulation (NMES) is a physical therapy intervention used to treat muscle weakness. NMES-elicited forces during therapy are correlated with strength gains. Patient discomfort limits NMES-elicited forces potentially compromising strength gains and the efficacy of this invention. The purpose of this study was to determine if NMES containing two different electrical stimulation pulse durations (200 or 500 µs) affected the knee extensor muscle torques subjects tolerated. Other NMES characteristics were identical in the two conditions: monophasic square-wave pulses; 75 pulses per second; and electrical stimulation train duration of one second. The primary dependent variable of interest was the percentage of maximum voluntary isometric contraction (MVIC) tolerated. The two pulse duration conditions were tested during a single session on the opposing lower extremities of 15 subjects. Subjects tolerated 49.3 ± 18.7% of MVIC torques in the 500-µs condition versus 44.5 ± 17.9% of MVIC torques in the 200-µs condition, which was a statistically significant difference (p = 0.02). Further research is needed to explore if the differences observed in this study would lead to clinically significant differences in strength gains and to see if the findings of this study can be generalized to other forms of NMES that contain other types of wave forms.