Proposed mechanism for sperm chromatin condensation/decondensation in the male rat

Condensation of sperm chromatin occurs after spermatozoa have left the caput epididymis and are in transit to the cauda epididymis, during which time large numbers of disulfide bonds are formed. The formation of these disulfide bonds requires the repeated oxidation of the cofactor, NAD(P)H. To date, the means by which this oxidation is achieved has yet to be elucidated. Spermatozoa lose the bulk of their cytoplasm prior to leaving the testis; and, as a result, any shuttle systems for removing and transferring reducing equivalents into the mitochondria are unlikely to be operational. In an apparent preparation for the loss of cytoplasm, however, the following events occur during spermatogenesis. First, androgen-binding protein (ABP) is produced by the Sertoli cells of the testis; second, high affinity binding sites for ABP are inserted into the membrane surrounding the nucleus; and third, a nuclear location is acquired for the enzyme, 3α-hydroxysteroid dehydrogenase (3α-HSD). We propose that after the loss of cytoplasm, the nuclear region of spermatozoa is directly accessible to constituents contained in the lumen of the caput epididymis. As a consequence, luminal ABP attaches itself to the nuclear membrane via its binding sites, and is internalized. After internalization, ABP exerts its principle function, which is to bind to luminal 5α-dihydrotestosterone (5α-DHT), thereby ensuring its availability to the enzyme, 3α-HSD. In the conversion of 5α-DHT to 3α-androstanediol (3α-Diol), NAD(P)H is oxidized. Spermatozoa that reach the cauda epididymis have fully condensed chromatin. In addition, the nuclear region retains appreciable amounts of 5α-DHT and 3α-Diol, both bound to ABP. During fertilization, the bound 3α-Diol is converted back to 5α-DHT, reducing equivalents are transferred to NAD(P)(+), and disulfide bonds are broken. IVF clinics report that spermatozoa with incompletely condensed chromatin have a low percentage of fertilization. If our proposed mechanism for chromatin condensation/decondensation is borne out by further research, IVF clinics might consider preincubating spermatozoa with 5α-DHT in order to increase the efficiency of fertilization

Everyday visual demands of people with low vision:A mixed methods real-life recording study

Research has demonstrated that low vison aids (LVAs) can have a positive impact on the functional sight of those living with sight loss. Step changes in technology are now enabling new wearable LVAs with greater potential than those available previously. For these novel devices to receive increased acceptance and therefore adoption by those with sight loss, visual task demands have to be understood more clearly in order to enable better alignment between device design and user requirements. The aim of this study was to quantify these requirements. Thirty-two participants aged 18 to 87 wore a spectacle-mounted video camera to capture and narrate all everyday situations in which they would use a "perfect" sight aid during 1 week. Captured scenes were analyzed through categorization and computational image analysis. Results showed large variation in activities and lifestyles. Participants reported no available sight aid or coping strategy for 57% of the recorded activities. Reading made up 49% of all recorded tasks, the other half comprising non-textual information. Overall, 75% of captured activities were performed ad hoc (duration of 0-5 minutes), 78% occurred indoors, 58% occurred at home, 48% were lit by natural light, 68% included the object of interest within reach, and 69% required a single focus plane only. Around half of captured objects of interest had a size of 2 degrees visual angle (2.08 logarithm of the minimum angle of resolution [logMAR]) or smaller. This study highlights the need for a sight aid that can make both textual and non-textual scenes accessible while offering flexibility to accommodate individual lifestyles

Gain Modulation by Corticostriatal and Thalamostriatal Input Signals during Reward-Conditioned Behavior.

The cortex and thalamus send excitatory projections to the striatum, but little is known about how these inputs, either individually or collectively, regulate striatal dynamics during behavior. The lateral striatum receives overlapping input from the secondary motor cortex (M2), an area involved in licking, and the parafascicular thalamic nucleus (PF). Using neural recordings, together with optogenetic terminal inhibition, we examine the contribution of M2 and PF projections on medium spiny projection neuron (MSN) activity as mice performed an anticipatory licking task. Each input has a similar contribution to striatal activity. By comparing how suppressing single or multiple projections altered striatal activity, we find that cortical and thalamic input signals modulate MSN gain and that this effect is more pronounced in a temporally specific period of the task following the cue presentation. These results demonstrate that cortical and thalamic inputs synergistically regulate striatal output during reward-conditioned behavior

Effects of human-induced alteration of groundwater flow on concentrations of naturally-occurring trace elements at water-supply wells

AbstractThe effects of human-induced alteration of groundwater flow patterns on concentrations of naturally-occurring trace elements were examined in five hydrologically distinct aquifer systems in the USA. Although naturally occurring, these trace elements can exceed concentrations that are considered harmful to human health. The results show that pumping-induced hydraulic gradient changes and artificial connection of aquifers by well screens can mix chemically distinct groundwater. Chemical reactions between these mixed groundwaters and solid aquifer materials can result in the mobilization of trace elements such as U, As and Ra, with subsequent transport to water-supply wells. For example, in the High Plains aquifer near York, Nebraska, mixing of shallow, oxygenated, lower-pH water from an unconfined aquifer with deeper, confined, anoxic, higher-pH water is facilitated by wells screened across both aquifers. The resulting higher-O2, lower-pH mixed groundwater facilitated the mobilization of U from solid aquifer materials, and dissolved U concentrations were observed to increase significantly in nearby supply wells. Similar instances of trace element mobilization due to human-induced mixing of groundwaters were documented in: (1) the Floridan aquifer system near Tampa, Florida (As and U), (2) Paleozoic sedimentary aquifers in eastern Wisconsin (As), (3) the basin-fill aquifer underlying the California Central Valley near Modesto (U), and (4) Coastal Plain aquifers of New Jersey (Ra). Adverse water-quality impacts attributed to human activities are commonly assumed to be related solely to the release of the various anthropogenic contaminants to the environment. The results show that human activities including various land uses, well drilling, and pumping rates and volumes can adversely impact the quality of water in supply wells, when associated with naturally-occurring trace elements in aquifer materials. This occurs by causing subtle but significant changes in geochemistry and associated trace element mobilization as well as enhancing advective transport processes

Mitochondrial 3 beta-hydroxysteroid dehydrogenase (HSD) is essential for the synthesis of progesterone by corpora lutea: An hypothesis

In mouse ovaries, the enzyme 3 beta-hydroxysteroid dehydrogenase (HSD) is distributed between microsomes and mitochondria. Throughout the follicular phase of the estrous cycle, the HSD activity in microsomes is predominant; whereas, after LH stimulation, HSD activity during the luteal phase is highest in the mitochondria. The current study examined whether or not LH stimulation always results in an increase in mitochondrial HSD activity. This was accomplished by measuring the HSD activity in microsomal and mitochondrial fractions from ovaries of pregnant mice. These animals have two peaks of LH during gestation, and one peak of LH after parturition. It was found that mitochondrial HSD activity was highest after each peak of LH. It is proposed that mitochondrial HSD is essential for the synthesis of high levels of progesterone. The increase in HSD activity in mitochondria after LH stimulation occurs because: 1) LH initiates the simultaneous synthesis of HSD and the cholesterol side-chain cleavage enzyme (P450scc); and, 2) HSD and P450scc bind together to form a complex, which becomes inserted into the inner membrane of the mitochondria. High levels of progesterone are synthesized by mitochondrial HSD because: 1) the requisite NAD+ cofactor for progesterone synthesis is provided directly by the mitochondria, rather than indirectly via the rate limiting malate-aspartate shuttle; and, 2) the end-product inhibition of P450scc by pregnenolone is eliminated because pregnenolone is converted to progesterone

K2p3.1 protein is expressed as a transmural gradient across the rat left ventricular free wall

Introduction: K 2p 3.1, also known as TASK-1, is a twin-pore acid-sensitive repolarizing K + channel, responsible for a background potassium current that significantly contributes to setting the resting membrane potential of cardiac myocytes. Inhibition of I K2p3.1 alters cardiac repolarization and is proarrhythmogenic. In this study, we have examined the expression of K 2p 3.1 and function of this channel in tissue and myocytes from across the left ventricular free wall. Methods and Results: Using fluorescence immunocytochemistry, the expression of K 2p 3.1 protein in myocytes from the subendocardial region was found to be twice (205% ± 13.5%) that found in myocytes from the subepicardial region of the left ventricle (100% ± 5.3%). The left ventricular free wall exhibited a marked transmural gradient of K 2p 3.1 protein expression. Western blot analysis confirmed significantly higher K 2p 3.1 protein expression in subendocardial tissue (156% ± 2.5%) than subepicardial tissue (100% ± 5.0%). However, there was no difference in K 2p 3.1 messenger RNA expression. Whole-cell patch clamp identified I K2p3.1 current density to be significantly greater in myocytes isolated from the subendocardium (7.66 ± 0.53 pA/pF) compared with those from the subepicardium (3.47 ± 0.74 pA/pF). Conclusions: This is the first study to identify a transmural gradient of K 2p 3.1 in the left ventricle. This gradient has implications for understanding ventricular arrhythmogenesis under conditions of ischemia but also in response to other modulatory factors, such as adrenergic stimulation and the presence of anesthetics that inhibits or activates this channel

Fragment screening reveals salicylic hydroxamic acid as an inhibitor of <em>Trypanosoma brucei</em> GPI GlcNAc-PI de-N-acetylase

The zinc-metalloenzyme GlcNAc-PI de-N-acetylase is essential for the biosynthesis of mature GPI anchors and has been genetically validated in the bloodstream form of Trypanosoma brucei, which causes African sleeping sickness. We screened a focused library of zinc-binding fragments and identified salicylic hydroxamic acid as a GlcNAc-PI de-N-acetylase inhibitor with high ligand efficiency. This is the first small molecule inhibitor reported for the trypanosome GPI pathway. Investigating the structure activity relationship revealed that hydroxamic acid and 2-OH are essential for potency, and that substitution is tolerated at the 4- and 5-positions

High resolution infrared spectra of bulge globular clusters: Liller~1 and NGC 6553

Using the NIRSPEC spectrograph at Keck II, we have obtained echelle spectra covering the range 1.5-1.8um for 2 of the brightest giants in Liller 1 and NGC 6553, old metal rich globular clusters in the Galactic bulge. We use spectrum synthesis for the abundance analysis, and find [Fe/H]=-0.3 +/- 0.2 and [O/H]=+0.3 +/- 0.2 dex. The composition of the clusters is similar to that of field stars in the bulge and is consistent with a sceanrio in which the clusters formed early, with rapid enrichment. We have dificulty achieveing a good fit to the spectrum of NGC 6553 using either the low or the high values recently reported in the literature, unless unusually large, or no alpha-element enhancements are adopted, respectively.Comment: To appear in the Astronomical Journal, March 200

Context-based generation of kinetic equations with SBMLsqueezer 1.3

The development of predictive, quantitative models constitutes a common task in today&#x2018;s systems biology. To obtain a mathematical model description for the simulation of gene-regulatory, signaling, and metabolic networks, kinetic equations are required for each reaction within the network. Deriving and assembling these formulas is a complicated, time-consuming, and error-prone process that requires knowledge about the structure of interactions, consistently choosing a rate law for each type of reaction, and assignment of appropriate units to all parameters. In many cases, thermodynamic dependencies between the parameters have to be taken into account. For multi compartment models, the concentration units of reacting species have to be converted into molecular amounts. Here we present version 1.3 of the program SBMLsqueezer that generates kinetic equations for SBML models based on the definition of the network&#x27;s topology and annotations of the elements therein, i.e., Systems Biology Ontology (SBO) and Minimal Information Requested In the Annotation of Models (MIRIAM) annotations

Spatial/Spectral Resolution of a Galactic Bulge K3 Giant Stellar Atmosphere via Gravitational Microlensing

We present two Keck HIRES spectra (R=40,000) of a bulge K3 giant taken on successive nights during the second caustic crossing of the binary microlensing event EROS BLG-2000-5. This caustic crossing served to effectively resolve the surface of the source star: the spectrum from the second night is dominated by the limb, while the spectrum from the first night is comprised of light from a broader range of radii. To demonstrate that the spectra are adequate to resolve the differences between them, we analyze the H-alpha line. The equivalent width is ~8% smaller on the second night, and the signal-to-noise ratio per resolution element (165 and 75 on the two nights respectively) is sufficient to show that the difference is approximately constant over the ~2A (~15 resolution element) extent of the line. The sign of the difference is in the expected direction since the limb is the coolest part of the star and therefore should have the weakest H-alpha. We invite atmosphere modelers to predict the difference spectrum from the entire spectral range 5500<lambda<7900A so that these predictions can be compared to our observations.Comment: 12 pages, 3 figures, Accepted by ApJ Letters 2001 Jan