Enhanced gross visualization of chicken Peyer's patch: novel staining technique applied to fresh tissue specimens

The ileal Peyer's patches (Pp), secondary gut-associated lymphoid tissue of the mucosal immune system, may serve as an important site for monitoring inflammatory and immunologic responses of the host against enteric pathogens. Chicken Pp are often difficult to observe grossly, and a simple technique to enhance visualization of the Pp is lacking. Therefore, we designed a novel staining method that is quick, easy, and accurate to aid in gross identification and recovery of the chicken Pp from fresh tissue specimens. Lower alimentary tracts were harvested from White Leghorn hens and commercial broilers. The ileocecocolic region was excised intact, flushed with deionized water to remove ingesta, and a dilute eosin-Y solution was infused. After 1 min, the eosin-Y was gently extruded. Modified-crystal violet (mCV) was then injected into the gastrointestinal segment, where on the lymphoid tissue area became apparent at the serosal surface. The distal ileal Pp was visible as a pale whitish pink ovoid-focalized area with surrounding gut tissue stained light purple. The exact Pp site could be delineated at the serosal and mucosal surface by gross assessment. Light microscopy evaluation of hematoxylin and eosin-stained tissue slides prepared from the excised Pp site revealed lymphoid tissue aggregations with multiple follicular units indicative of Pp. The novel eosin-Y + mCV staining technique promotes rapid identification and accurate recovery of chicken Pp lymphoid tissue from fresh tissue specimens.     

Cellular and extracellular vesicular origins of miRNAs within the bovine ovarian follicle

The ovarian follicle components must provide an ideal environment to ensure the success of reproductive processes, and communication between follicular cells is crucial to support proper oocyte growth. Recently, it has been demonstrated that the presence of extracellular vesicles (EVs) carrying microRNAs (miRNAs) in follicular fluid represents an important autocrine and paracrine communication mechanism inside the ovarian follicle. In this study, we tested the hypothesis that the miRNA content of EVs isolated from ovarian follicular (granulosa and cumulus–oocyte complexes) cell‐conditioned culture media is dependent upon cell type. We initially screened bovine granulosa cells (GCs) and cumulus–oocyte complexes (COCs), as well as their derived EVs for 348 miRNAs using real‐time PCR, and detected 326 miRNAs in GCs and COCs cells and 62 miRNAs in GCs and COCs EVs. A bioinformatics analysis of the identified cell‐specific and differentially expressed miRNAs predicted that they likely modulate important cellular processes, including signalling pathways such as the PI3K‐Akt, MAPK and Wnt pathways. By investigating the origins of miRNAs within the follicular fluid, the results of this study provide novel insights into follicular miRNA content and intercellular communication that may be of invaluable use in the context of reproductive technologies, diagnostic of ovarian‐related diseases and/or the identification of biomarkers for oocyte and embryo quality.     

Effect of hCG on Early Luteal Serum Progesterone Concentrations in PG600‐Treated Gilts

Gilt oestrus and ovulation responses to injection of a combination of equine chorionic gonadotrophin (eCG) and human chorionic gonadotrophin (hCG) (PG600) can be unpredictable, possibly reflecting inadequate circulating LH activity. The objective of this study was to determine the effect of PG600 followed by supplemental hCG on gilt ovarian responses. In experiment 1, 212 Hypor gilts (160 day of age) housed on two farms in Spain received intramuscular (i.m.) injections of PG600 (n = 47), or PG600 with an additional 200 IU hCG injected either concurrently (hCG‐0; n = 39), or at 24 h (hCG‐24; n = 41) or 48 h (hCG‐48; n = 45) after PG600. A further 40 gilts served as non‐injected controls. Ovulation responses were determined on the basis of initial blood progesterone concentrations being <1 ng/ml and achieving >5 ng / ml 10 d after the PG600 injection. The incidence of ovulating gilts having progesterone concentrations >30 ng/ml were recorded. During the study period, 10% of control gilts ovulated whereas 85–100% of hormone‐treated gilts ovulated. There were no significant differences among hormone groups for proportions of gilts ovulating. The proportions of gilts having circulating progesterone concentrations >30 ng/ml were increased (p ≤ 0.02) in all hCG treated groups compared with the PG600 group. In experiment 2, a total of 76 Hypor gilts at either 150 or 200 days of age were injected with PG600 (n = 18), 400 IU eCG followed by 200 IU hCG 24 h later (n = 20), PG600 followed by 100 IU hCG 24 h later (n = 17), or 400 IU eCG followed by 300 IU hCG 24 h later (n = 21). Blood samples were obtained 10 days later for progesterone assay. There were no effects of treatment or age on incidence of ovulation, but fewer 150‐day‐old gilts treated with PG600 or 400 IU eCG followed by 200 IU hCG had progesterone concentrations >30 ng / ml. We conclude that hCG treatment subsequent to PG600 treatment will generate a higher circulating progesterone concentration, although the effect is not evident in older, presumably peripubertal, gilts. The mechanism involved and implications for fertility remain to be determined.     

Penny ice cap cores, baffin island, canada, and the wisconsinan foxe dome connection: two states of hudson bay ice cover

Ice cores from Penny Ice Cap, Baffin Island, Canada, provide continuous Holocene records of oxygen isotopic composition (delta18O, proxy for temperature) and atmospheric impurities. A time scale was established with the use of altered seasonal variations, some volcanic horizons, and the age for the end of the Wisconsin ice age determined from the GRIP and GISP2 ice cores. There is pre-Holocene ice near the bed. The change in delta18O since the last glacial maximum (LGM) is at least 12.5 per mil, compared with an expected value of 7 per mil, suggesting that LGM ice originated at the much higher elevations of the then existing Foxe Dome and Foxe Ridge of the Laurentide Ice Sheet. The LGM delta18O values suggest thick ice frozen to the bed of Hudson Bay.     

The structure of the upper atmosphere of mars: In situ accelerometer measurements from mars global surveyor

The Mars Global Surveyor (MGS) z-axis accelerometer has obtained over 200 vertical structures of thermospheric density, temperature, and pressure, ranging from 110 to 170 kilometers, compared to only three previous such vertical structures. In November 1997, a regional dust storm in the Southern Hemisphere triggered an unexpectedly large thermospheric response at mid-northern latitudes, increasing the altitude of thermospheric pressure surfaces there by as much as 8 kilometers and indicating a strong global thermospheric response to a regional dust storm. Throughout the MGS mission, thermospheric density bulges have been detected on opposite sides of the planet near 90 degreesE and 90 degreesW, in the vicinity of maximum terrain heights. This wave 2 pattern may be caused by topographically-forced planetary waves propagating up from the lower atmosphere.     

A novel and efficient method for identifying cotton leafroll dwarf virus infection in upland cotton (Gossypium hirsutum)

We report the development of a novel and more efficient, rapid, cost-effective and simple technique than current PCR-based identification methods for screening cotton (Gossypium hirsutum) plants for the presence of cotton leafroll dwarf virus (CLRDV). This protocol takes advantage of the PACE (PCR Allele Competitive Extension) system and uses PCR amplification of cDNA, coupled with sequence-specific fluorescent probes to differentiate between infected and uninfected cotton plants. This procedure has the potential for application in detection of other RNA viruses in a variety of other crops, by using primers specific for the RNA-dependent RNA polymerase (RdRP) gene and a widely conserved housekeeping gene in the host organism; in this case, the G. hirsutum polyubiquitin gene (GhUB).     

Analysis of bromide ion in wine by ion selective electrode.

The use of the bromide ion selective electrode for the determination of bromide ion in wine has been found to be rapid and reliable. The method has been used for still wines and carbonated wines and is applicable to all wines regardless of their country of origin. The method consists of treating a 50 ml aliquot of wine with 2 ml each of 3.75M H3PO4, saturated KNO3, and 1M CuSO4. After 10 min the electrodes are immersed in the samples and a millivolt reading is obtained. One hundred mul 500 ppm bromide ion standard is added and the millivolt reading is taken. Bromide ion concentration in the wine equals (Cdelta x 1)/((antilog deltaE/S)-1) where Cdelta equals 1, deltaE equals the change in potential expressed in millivolts, and S equals the electrode slope.     

Sicklepod (Senna obtusifolia) seed processing and potential utilization

Sicklepod (Senna obtusifolia) is a leguminous plant that infests soybean fields in the southeastern United States. Its seeds contain a variety of toxic, highly colored compounds, mainly anthraquinones together with a small amount of fat. These compounds contaminate and lower the quality of soybean oil when inadequately cleaned soybean seed from this area is processed. The sorting of sicklepod seed from a soybean harvest is an additional economic burden on the farmer beyond the cost of proper disposal of the weed seed to avoid worsening field infestation. Fortunately, sicklepod seed also contains substantial amounts of carbohydrates and proteins. These edible components when freed from anthraquinones have a market in pet food as well as potential in human foods because of the high galactomannan ratio of the polysaccharides. Sicklepod seed was dehulled, and the ground endosperm was defatted, followed by sequential solvent extraction of the defatted seed meal to isolate the anthraquinones, carbohydrates, and protein components into their respective classes. Each class of isolate was spectroscopically identified.