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Oocyte morphology and estrogen concentrations following a reduction in progesterone in beef cattle

Low dosages of progestogens promote persistent follicles, high systemic estrogen and low fertility. The objectives of this study were to determine effects of a reduction in progesterone on (1) morphology of oocytes and intrafollicular concentrations of estradiol. Cows on low progesterone (n = 12) received used intravaginal progesterone inserts on d 4 after estrus and prostaglandin (PG) F2alpha (25 mg, i.m.) on d 6. Control animals (n = 12) received saline on d 6. The oocyte and follicular fluid were recovered from the largest follicle on d 8 or d 10.;Serum estradiol was lower during d 4--6 but greater (P \u3c .01) during d 7--10 in cows treated with progesterone inserts and PGF 2alpha while the largest follicle was larger in treated cows on day 10 only (14 vs. 12 mm; P \u3c .05). Intrafollicular concentrations of estrogen were greater in treated than in control cows (990 +/- 87 vs 191 +/- 106; P \u3c .01). Progesterone in follicular fluid (mean = 42 ng/ml) did not differ. Oocytes were observed in oocyte nuclear stage I in the control group on d 8. All other oocytes were in nuclear stage II. In addition, the degree of clumping of mitochondria, the percentage of intact cumulus cell processes and percentage of normally shaped mitochondria was greater in oocytes from d 8 control cows than in all other groups.;Changes in concentrations of estradiol and oocyte morphology typically associated with the preovulatory period had occurred within 2 d after a reduction in progesterone, even when low peripheral concentrations of progesterone were maintained. These earliest stages of oocyte maturation occurred in response to a reduction in progesterone. Similar changes in oocyte morphology were observed in control animals by d 10 of the estrous cycle, probably representing the onset of atresia

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Safety and Health Administration proposed rule on Proximity Detection Systems for Continuou

Mouse Estrous Cycle Identification Tool and Images

The efficiency of producing timed pregnant or pseudopregnant mice can be increased by identifying those in proestrus or estrus. Visual observation of the vagina is the quickest method, requires no special equipment, and is best used when only proestrus or estrus stages need to be identified. Strain to strain differences, especially in coat color can make it difficult to determine the stage of the estrous cycle accurately by visual observation. Presented here are a series of images of the vaginal opening at each stage of the estrous cycle for 3 mouse strains of different coat colors: black (C57BL/6J), agouti (CByB6F1/J) and albino (BALB/cByJ). When all 4 stages (proestrus, estrus, metestrus, and diestrus) need to be identified, vaginal cytology is regarded as the most accurate method. An identification tool is presented to aid the user in determining the stage of estrous when using vaginal cytology. These images and descriptions are an excellent resource for learning how to determine the stage of the estrous cycle by visual observation or vaginal cytology

Studies on Dibenzylamines as Inhibitors of Venezuelan Equine Encephalitis Virus

Alphaviruses are arthropod-transmitted members of the Togaviridae family that can cause severe disease in humans, including debilitating arthralgia and severe neurological complications. Currently, there are no approved vaccines or antiviral therapies directed against the alphaviruses, and care is limited to treating disease symptoms. A phenotypic cell-based high-throughput screen was performed to identify small molecules that inhibit the replication of Venezuelan Equine Encephalitis Virus (VEEV). The compound, 1-(2,3-dihydrobenzo[b][1,4]dioxin-6-yl)-N-(3-fluoro-4-methoxybenzyl)ethan-1-amine (1), was identified as a highly active, potent inhibitor of VEEV with an effective concentration for 90% inhibition of virus (EC90) of 0.89 μM and 7.49 log reduction in virus titers at 10 μM concentration. These data suggest that further investigation of compound 1 as an antiviral therapeutic against VEEV, and perhaps other alphaviruses, is warranted. Experiments suggested that the antiviral activity of compound 1 is directed at an early step in the VEEV replication cycle by blocking viral RNA and protein synthesis

Delayed Stellar Mass Assembly in the Low Surface Brightness Dwarf Galaxy KDG215

We present HI spectral line and optical broadband images of the nearby low surface brightness dwarf galaxy KDG215. The HI images, acquired with the Karl G. Jansky Very Large Array (VLA), reveal a dispersion dominated ISM with only weak signatures of coherent rotation. The HI gas reaches a peak mass surface density of 6 M$_{\odot}$ pc$^{-2}$ at the location of the peak surface brightness in the optical and the UV. Although KDG215 is gas-rich, the H$\alpha$ non-detection implies a very low current massive star formation rate. In order to investigate the recent evolution of this system, we have derived the recent and lifetime star formation histories from archival Hubble Space Telescope images. The recent star formation history shows a peak star formation rate $\sim$1 Gyr ago, followed by a decreasing star formation rate to the present day quiescent state. The cumulative star formation history indicates that a significant fraction of the stellar mass assembly in KDG215 has occurred within the last 1.25 Gyr. KDG215 is one of only a few known galaxies which demonstrates such a delayed star formation history. While the ancient stellar population (predominantly red giants) is prominent, the look-back time by which 50% of the mass of all stars ever formed had been created is among the youngest of any known galaxy.Comment: Accepted for publication in the Astrophysical Journal Letter

Conserving, Distributing and Managing Genetically Modified Mouse Lines by Sperm Cryopreservation

Sperm from C57BL/6 mice are difficult to cryopreserve and recover. Yet, the majority of genetically modified (GM) lines are maintained on this genetic background.Reported here is the development of an easily implemented method that consistently yields fertilization rates of 70+/-5% with this strain. This six-fold increase is achieved by collecting sperm from the vas deferens and epididymis into a cryoprotective medium of 18% raffinose (w/v), 3% skim milk (w/v) and 477 microM monothioglycerol. The sperm suspension is loaded into 0.25 mL French straws and cooled at 37+/-1 degrees C/min before being plunged and then stored in LN(2). Subsequent to storage, the sperm are warmed at 2,232+/-162 degrees C/min and incubated in in vitro fertilization media for an hour prior to the addition of oocyte cumulus masses from superovulated females. Sperm from 735 GM mouse lines on 12 common genetic backgrounds including C57BL/6J, BALB/cJ, 129S1/SvImJ, FVB/NJ and NOD/ShiLtJ were cryopreserved and recovered. C57BL/6J and BALB/cByJ fertilization rates, using frozen sperm, were slightly reduced compared to rates involving fresh sperm; fertilization rates using fresh or frozen sperm were equivalent in all other lines. Developmental capacity of embryos produced using cryopreserved sperm was equivalent, or superior to, cryopreserved IVF-derived embryos.Combined, these results demonstrate the broad applicability of our approach as an economical and efficient option for archiving and distributing mice

Subsequent Surgery After Revision Anterior Cruciate Ligament Reconstruction: Rates and Risk Factors From a Multicenter Cohort

BACKGROUND: While revision anterior cruciate ligament reconstruction (ACLR) can be performed to restore knee stability and improve patient activity levels, outcomes after this surgery are reported to be inferior to those after primary ACLR. Further reoperations after revision ACLR can have an even more profound effect on patient satisfaction and outcomes. However, there is a current lack of information regarding the rate and risk factors for subsequent surgery after revision ACLR. PURPOSE: To report the rate of reoperations, procedures performed, and risk factors for a reoperation 2 years after revision ACLR. STUDY DESIGN: Case-control study; Level of evidence, 3. METHODS: A total of 1205 patients who underwent revision ACLR were enrolled in the Multicenter ACL Revision Study (MARS) between 2006 and 2011, composing the prospective cohort. Two-year questionnaire follow-up was obtained for 989 patients (82%), while telephone follow-up was obtained for 1112 patients (92%). If a patient reported having undergone subsequent surgery, operative reports detailing the subsequent procedure(s) were obtained and categorized. Multivariate regression analysis was performed to determine independent risk factors for a reoperation. RESULTS: Of the 1112 patients included in the analysis, 122 patients (11%) underwent a total of 172 subsequent procedures on the ipsilateral knee at 2-year follow-up. Of the reoperations, 27% were meniscal procedures (69% meniscectomy, 26% repair), 19% were subsequent revision ACLR, 17% were cartilage procedures (61% chondroplasty, 17% microfracture, 13% mosaicplasty), 11% were hardware removal, and 9% were procedures for arthrofibrosis. Multivariate analysis revealed that patients aged <20 years had twice the odds of patients aged 20 to 29 years to undergo a reoperation. The use of an allograft at the time of revision ACLR (odds ratio [OR], 1.79; P = .007) was a significant predictor for reoperations at 2 years, while staged revision (bone grafting of tunnels before revision ACLR) (OR, 1.93; P = .052) did not reach significance. Patients with grade 4 cartilage damage seen during revision ACLR were 78% less likely to undergo subsequent operations within 2 years. Sex, body mass index, smoking history, Marx activity score, technique for femoral tunnel placement, and meniscal tearing or meniscal treatment at the time of revision ACLR showed no significant effect on the reoperation rate. CONCLUSION: There was a significant reoperation rate after revision ACLR at 2 years (11%), with meniscal procedures most commonly involved. Independent risk factors for subsequent surgery on the ipsilateral knee included age <20 years and the use of allograft tissue at the time of revision ACLR