T-cell lymphoma infiltrating the uterus and ovaries of a Golden Retriever: a case report.

BACKGROUND: To the best of our knowledge, this is the first report of female genital system infiltration of T-cell lymphoma in veterinary literature. CASE PRESENTATION: A 1.5-year-old, intact female Golden Retriever was referred due to melena and hyporexia that lasted for three weeks. Fever (40.5℃), tachycardia, tachypnoea, pale mucous membranes, and purulent vaginal discharge were identified on physical examination. Blood analyses revealed leucocytosis, anaemia, hypoalbuminemia, and increased lactate and C-reactive protein levels. On abdominal radiography, the small intestine was moderately deviated because of an oval-shaped mass (13 cm × 8.7 cm) located in the mid-abdomen. An enlarged tubular-shaped structure that had the opacity of soft tissue located in dorsal to the bladder to the middle of the abdomen, and an oval-shaped mass (5.28 cm × 3.26 cm), which was suspected to be a medial iliac lymph node located at the sixth to seventh lumbar level. Abdominal ultrasonography revealed gas and fluid in the lumen of the uterine horn with a severely thickened wall, round enlarged lymph nodes around the genitourinary system, and free fluid in the abdominal cavity. Based on these results, pyometra was suspected, and an exploratory laparotomy was performed for ovariohysterectomy. The resected ovary and uterus were macroscopically hypertrophied. Histopathological examination of the ovary and uterus revealed neoplastic proliferation of large round cells with strong immunoreactivity for CD3, indicating T-cell lymphoma. Therefore, the young dog was diagnosed with genital lymphoma. CONCLUSIONS: The present report describes T-cell lymphoma infiltrating the uterus and ovaries in a young dog, which is rarely diagnosed and could aid in the differential diagnosis of genital diseases in young dogs.

Scalable graph neural network for NMR chemical shift prediction.

Graph neural networks (GNNs) have been proven effective in the fast and accurate prediction of nuclear magnetic resonance (NMR) chemical shifts of a molecule. Existing methods, despite their effectiveness, suffer from high space complexity and are therefore limited to relatively small molecules. In this work, we propose a scalable GNN for NMR chemical shift prediction. To reduce the space complexity, we sparsify the graph representation of a molecule by regarding only heavy atoms as nodes and their chemical bonds as edges. To better learn from the sparsified graph representation, we improve the message passing and readout functions in the GNN. For the message passing function, we adapt the attention mechanism and residual connection to better capture local information around each node. For the readout function, we use both node-level and graph-level embeddings as the local and global information to better predict node-level chemical shifts. Through the experimental investigation using 13C and 1H NMR datasets, we demonstrate that the proposed method yields higher prediction accuracy and is more scalable to large molecules having many heavy atoms.

Factors affecting reproductive outcomes in lactating dairy cows that undergo presynchronization-Ovsynch and successive resynchronization programs.

This study was designed to identify the factors affecting reproductive outcomes in lactating dairy cows undergoing Presynchronization-Ovsynch and successive resynchronization programs. Data were collected from 1,458 cows on 11 dairy farms, regarding nutrition, health, reproduction, and specifically reproductive programs used for the first timed artificial insemination (TAI; Modified Presynch-Ovsynch or Modified Double-Ovsynch) and for re-inseminations (Ovsynch or PreGnRH-Ovsynch); as well as the weather. Body condition score (BCS) and timing of TAI affected (P < 0.01) the probability of pregnancy per AI after first TAI. Cows with a BCS ≥3.25 were more likely (P < 0.01) to be pregnant at 31 (odds ratio [OR]: 1.85) and 49 (OR: 1.93) days after the first TAI than cows with BCS ≤2.75. Cows inseminated during months of heat stress (June to September) were less likely (P < 0.01) to be pregnant at 31 (OR: 0.74) and 49 (OR: 0.68) days after first TAI than those inseminated during months of no heat stress (October to May). Cows that had a pre- or postpartum disorder were more likely to lose their pregnancy (OR: 2.17, P < 0.01) than those that did not. Cows inseminated ≥76 days after calving (OR: 1.67, P < 0.1) or during the months of heat stress (OR: 2.02, P < 0.05) were also more likely to lose pregnancy than cows inseminated ≤75 days after calving or during the months of no heat stress. Following successive resynchronizations, cows with a third (OR: 1.32, P < 0.1) or fourth (OR: 1.68, P < 0.05) TAI were more likely to be pregnant than those with a second TAI. Cows with BCS ≥3.25 at the beginning of resynchronization were more likely (OR: 1.49, P < 0.05) to be pregnant than cows with BCS ≤2.75. Finally, the likelihood of being pregnant by 210 days postpartum was positively associated with a favorable BCS, but negatively associated with the presence of a pre- or postpartum disorder and a larger number of days to first TAI following calving (≥76 vs. ≤75 days). In conclusion, high BCS has beneficial effects during the entire reproductive period, whereas TAI during heat stress, the presence of a pre- or postpartum disorder, a larger number of days to first TAI following calving, and TAI number (second or ≥ fifth) adversely affects the outcomes associated with pregnancy during the early or later breeding periods in lactating dairy cows undergoing Presynchronization-Ovsynch and successive resynchronization programs.

Evaluation of Dose-Dependent Obesity and Diabetes-Related Complications of Water Chestnut (Fruit of Trapa japonica) Extracts in Type II Obese Diabetic Mice Induced by 45% Kcal High-Fat Diet.

Background and Objectives: The currently used pharmacological agents for metabolic disorders such as type II diabetes have several limitations and adverse effects; thus, there is a need for alternative therapeutic drugs and health functional foods. Materials and Methods: This study investigated the pharmacological effects of water chestnut (fruit of Trapa japonica) extracts (WC: 50-200 mg/kg) for type II diabetes using a 45% Kcal high-fat diet (HFD)-fed type II obese diabetic mice model for a period of 84 days, and the effects were compared to those of metformin (250 mg/kg). Results: Increases in body weight, serum biochemical indices such as triglycerides, low-density lipoprotein, and blood urea nitrogen, increases in antioxidant defense system enzymes such as catalase, superoxide dismutase, and glutathione, and mRNA expressions (such as AMPKα1 and AMPKα2) in the liver tissue and mRNA expressions (such as AMPKα2 mRNA, leptin, and C/EBPα) in the adipose tissue were observed in the HFD control group. The WC (50 mg/kg)-administered group showed no significant improvements in diabetic complications. However, HFD-induced obesity and diabetes-related complications such as hyperlipidemia, diabetic nephropathy, nonalcoholic fatty liver disease (NAFLD), oxidative stress, activity of antioxidant defense systems, and gene expressions were significantly and dose-dependently inhibited and/or normalized by oral administration of WC (100 mg/kg and 200 mg/kg), particularly at a dose of 100 mg/kg. Conclusions: The results of this study suggest that WC at an appropriate dose could be used to develop an effective therapeutic drug or functional food for type II diabetes and various associated complications, including NAFLD.

Selective use of a modified pre-synchronization-Ovsynch and resynchronization reproductive strategy in dairy herds: A field application study.

We aimed to evaluate whether a modified pre-synchronization-Ovsynch (MPO) and resynchronization method, adjusted for the timing of biweekly reproductive consulting visits, could be implemented successfully under field conditions in dairy herds. Forty-five days after calving (calving: Day 0), 1,182 cows were assigned randomly to Ovsynch, MPO or control groups. (1) Cows were administered GnRH on Day 65, PGF2α on Day 72 and GnRH 56 hr later, followed by timed artificial insemination (TAI) 16 hr later (Ovsynch, n = 380). (2) Cows with a CL were administered PGF2α (n = 204) and cows without were administered GnRH (n = 199) on Day 45. Then, each cow was administered PGF2α on Day 55 (9 to 12 days later), GnRH on Day 58 and underwent Ovsynch 7 days later (selective MPO [sMPO] n = 403). (3) The remaining cows underwent AI when oestrus was detected (EDAI, n = 399). The probabilities of pregnancy per AI 30 and 45 days after the first AI were higher in the sMPO group (odds ratios 1.99 and 1.92, p < .0001) than in the EDAI group. In addition, the probability of pregnancy by 105 days postpartum tended to be higher (p < .1) in the sMPO group than in the EDAI and Ovsynch groups, which shortened the mean interval from calving to pregnancy in the sMPO group (3.4 and 4.2 days, respectively) than in the EDAI and Ovsynch groups. In the sMPO group, the interval between the first and second treatments during pre-synchronization did not affect the probability of pregnancy per AI after the first AI. The probability of pregnancy per AI after the second AI did not differ between the Ovsynch, sMPO and EDAI groups. In conclusion, the sMPO programme, adjusted for the timing of biweekly reproductive consulting visits, followed by resynchronization, could be successfully adopted in field conditions, and is associated with good reproductive performance in dairy herds.

Seasonal characteristics of atmospheric water-soluble organic nitrogen in PM2.5 in Seoul, Korea: Source and atmospheric processes of free amino acids and aliphatic amines.

The seasonal characteristics of atmospheric water-soluble organic nitrogen (WSON) in particulate matter with a diameter of 2.5 µm or smaller (PM2.5) were analyzed focusing on sources and atmospheric processing. Daily collected samples over 23 h (10:00-9:00) from 7 August 2018 to 31 December 2019 on quartz filters with a high-volume sampler at the Korea Institute of Science and Technology (KIST) in Seoul were considered. The most common species in the Seoul atmosphere included Glycine (5.45 ± 9.81 ng/m3) among free amino acids (FAAs) and trimethylamine (TMA) (5.35 ± 3.80 ng/m3) among aliphatic amines (AAs). The top 10 WSON species (93.6% of all WSON species) were categorized into three groups based on correlation analysis considering meteorological data, (e.g., temperature, rainfall, relative humidity (RH), wind speed) gaseous pollutants (e.g., SO2, CO, NO2) and mass concentration of PM10 and PM2.5. Those three groups are G1 (Glycine, Alanine, and Threonine), G2 (Gln Glutamine, Lys Lysine, and Glutamic acid) and G3 (Trimethylamine (TMA), dimethylamine (DMA), and methylamine (MA)), where G1, G2 and G3 accounted for 31.1%, 8.8% and 51.1%, respectively, of the total species. Among these three groups, G1 and G3 are from combustion sources, and G2 shows secondary features generated by photochemical reactions involving ozone. Although both G1 and G3 exhibited features influenced by combustion sources, the AA species (TMA, DMA, and MA) in G3 demonstrated typical features enhanced under high-humidity conditions, suggesting not only primary sources but also secondary formation at the local scale influence to the AA in G3 group. Based on long-term measurements more than a year, our findings suggest that complex and diverse sources of atmospheric WSON are in Seoul, Korea both from primary and secondary, which may affect its environmental, climate and health.

Impacts of secondary aerosol formation and long range transport on severe haze during the winter of 2017 in the Seoul metropolitan area.

Severe haze episodes occur frequently in the Seoul Metropolitan Area (SMA) and throughout East Asian countries, especially during the winter and early spring. We investigated the sources and chemistry of particulate matter (PM) during three winter haze episodes in Seoul that occurred between January 1st and February 10th in 2017 using a high resolution time-of-flight aerosol mass spectrometer (HR-ToF-AMS) and positive matrix factorization (PMF) analysis. The average concentration of sub-micrometer aerosol (PM1 = NR-PM1 + black carbon (BC)) was 32.6 µg/m3, which was composed of 42% organics, 27% nitrate, 11% sulfate, 13% ammonium and 4% BC by mass. Six distinct sources of organic aerosol (OA) were identified: vehicle emitted hydrocarbon-like OA (HOA), cooking OA (COA), biomass burning OA (BBOA), and 3 different types of secondary OA (SOA) with varying degrees of oxidation and temporal trends. The nitrate mass fraction increased during the three haze episodes, with nitrate accounting for 27-33% of PM1 mass. Enhanced nitrate concentrations and higher nitrate oxidation ratios (NOR), despite lower enhancement in relative humidity (RH) than the low PM loading period, suggest that regional transport of nitrate contributed to the nitrate mass during the haze periods. Lower HOA and COA concentrations during the high PM loading periods further confirm that local emissions or stagnant meteorological conditions were not the main reason for the severe haze. Residence time analysis (RTA), concentration field analysis (CFA), and column-CFA results from FLEXPART also showed that the measurement period was accompanied by atmospheric transport of nitrate, sulfate, and ammonium from eastern China. Nevertheless, we found that NO2, a precursor of nitrate, was predominantly from local emissions. These findings suggest that nitrate in Seoul is not only locally formed, but also transported from upwind areas.

Lateral Flank Approach for Ovariohysterectomy in a Lion (Panthera leo) with a Ruptured Pyometra.

An 8.5-year-old intact female lion (Panthera leo) with a history of vomiting, lethargy, and anorexia was referred to our institution. On physical examination, the lion weighed 180 kg and had a rectal temperature of 40 °C. Blood analysis showed mild neutrophilic leukocytosis, and abdominal ultrasonography revealed an enlarged uterus with echogenic fluid. Based on the clinical signs and hematologic and ultrasonographic findings, the lion was tentatively diagnosed with pyometra. Ovariohysterectomy was performed using the lateral flank approach to avoid complications that can occur with ventral celiotomy. Surgery was performed successfully despite unexpected rupture of the uterus which had occurred before the surgery. The lion recovered uneventfully and continued to do well at the 1 year follow up after surgery. To the best of our knowledge, this is the first reported case of ovariohysterectomy with the lateral flank approach in a lion with pyometra. Our report suggests that ovariohysterectomy with the lateral flank approach could be considered as an alternative method for pyometra in wild big cats with risk of complications at the surgical site.

Anti-inflammatory effects of Capparis ecuadorica extract in phthalic-anhydride-induced atopic dermatitis of IL-4/Luc/CNS-1 transgenic mice.

CONTEXT: The natural products derived from Capparis ecuadorica H.H. Iltis (Capparaceae) could have great potential for anti-inflammation since they inhibited the inflammatory response in lipopolysaccharide (LPS)-stimulated RAW 264.7 cells. OBJECT: This study investigated the anti-inflammatory effects and related mechanism of methanol extract of C. ecuadorica leaves (MCE) during atopic dermatitis (AD) responses. MATERIALS AND METHODS: Alterations in the phenotypical markers for AD, luciferase signal, iNOS-mediated COX-2 induction pathway, and inflammasome activation were analysed in non-Tg (n = 5) and 15% phthalic anhydride (PA) treated IL-4/Luc/CNS-1 transgenic (Tg) HR1 mice (n = 5 per group), subsequent to treatment with acetone-olive oil (AOO), vehicle (DMSO) and two dose MCE (20 and 40 mg/kg) three times a week for 4 weeks. RESULTS: MCE treatment reduced the intracellular ROS level (48.2%), NO concentration (7.1 mmol/L) and inflammatory cytokine expressions (39.1%) in the LPS-stimulated RAW264.7 cells. A significant decrease was detected for ear thickness (16.9%), weight of lymph node (0.7 mg), IgE concentration (1.9 µg/mL), and epidermal thickness (31.8%) of the PA + MCE treated Tg mice. MCE treatment induced the decrease of luciferase signal derived from the IL-4 promoter and the recovery of the IL-4 downstream regulator cytokines. PA + MCE treated Tg mice showed decreasing infiltration of mast cells (42.5%), iNOS-mediated COX-2 induction pathway, MAPK signalling pathway and inflammasome activation in the ear tissue. CONCLUSIONS: These findings provide the first evidence that MCE may have great potential to suppress chemical-induced skin inflammation through the suppression of IL-4 cytokine and the iNOS-mediated COX-2 induction pathway, and activation of inflammasome.

Reproductive performance following a modified Presynch-Ovsynch, Double-Ovsynch, or conventional reproductive management program in Korean dairy herds.

We aimed to compare the reproductive performance of dairy cattle following a modified Presynch-Ovsynch, Double-Ovsynch, or conventional reproductive management program (CRMP). On Day 30 (±5) after calving (calving = Day 0), 960 lactating cows were assigned randomly to one of two presynchronization treatments, before the synchronization of ovulation (Ovsynch) or CRMP without presynchronization. Cows were administered prostaglandin F2α (PGF2α) on Days 38 (±5) and 52, and gonadotropin-releasing hormone (GnRH) on Day 55 (Presynch-G-Ovsynch group, n = 333); or GnRH on Day 45 (±5), PGF2α on Day 52, and GnRH again on Day 55 (Double-Ovsynch group, n = 307). Thereafter, each cow underwent Ovsynch 7 days later: GnRH on Day 62, PGF2α on Day 69, and GnRH again 56 h later, followed by timed artificial insemination (AI) 16 h later. The remaining cows underwent AI when estrus was detected or Ovsynch (CRMP group, n = 320). In a subset of cows (each n = 40) in the two presynchronization groups, blood collections and ovarian ultrasonography were performed on Days 30 (±4), 52, 62, and 69, and uterine cytology on Days 30 (±4) and 52. The probabilities of pregnancy per AI 32 and 60 days after the first AI were higher in the Presynch-G-Ovsynch (odds ratio [OR]: 1.89 and 1.81, P < 0.01) and Double-Ovsynch (OR: 1.63 and 1.60, P < 0.05) groups than in the CRMP group. The likelihood of pregnancy by 210 days postpartum was higher (P < 0.05) in Presynch-G-Ovsynch (hazard ratio [HR]: 1.29) and Double-Ovsynch (HR: 1.31) groups than in the CRMP group, whereas the least square mean number of inseminations per conception was lower (P < 0.05) in Presynch-G-Ovsynch (1.8) and Double-Ovsynch (1.8) groups than in the CRMP group (2.1). The percentages of cows with serum progesterone concentrations ≥1.0 ng/mL or with a corpus luteum (CL) did not differ between the groups (P > 0.1) on Days 30, 62, and 69, but were lower (P < 0.05) on Day 52 in the Presynch-G-Ovsynch than in the Double-Ovsynch groups. However, the diameter of the dominant follicles, the proportion of neutrophils in uterine cytological samples, and the prevalence of an accessory CL on Day 69 did not differ (P > 0.1) between the two groups. In conclusion, both the Presynch-G-Ovsynch and Double-Ovsynch programs improved reproductive performance vs. CRMP in smallholder Korean dairy herds, and there were no differences in the ovarian endocrine and structural dynamics, or uterine health, between the two programs involving presynchronization.

PET imaging of HER2 expression with an 18F-fluoride labeled aptamer.

BACKGROUND/PURPOSE: Aptamers are oligonucleotide or peptide molecules that bind to a target molecule with high affinity and specificity. The present study aimed to evaluate the target specificity and applicability for in vivo molecular imaging of an aptamer labeled with a radioisotope. METHODS: The human epidermal growth factor receptor 2 (HER2/ErbB2) aptamer was radiolabeled with 18F-fluoride. HER2-positive tumor cell uptake of the aptamer was evaluated in comparison to negative controls by flow cytometry and confocal microscopy. Using 18F-labeled HER2-specific aptamer positron emission tomography (PET), in vivo molecular images of BT474 tumor-bearing mice were taken at 60, 90 and 120 minutes after injection. RESULTS: In flow cytometric analysis, HER2 aptamer showed strong binding to HER2-positive BT474 cells, while binding to HER2-negative MDA-MB231 cells was quite low. Likewise, in confocal microscopic images, the aptamer was bound to HER2-positive breast cancer cells, with minimal binding to HER2-negative cells. In vivo PET molecular imaging of BT474 tumor-bearing mice revealed significant higher uptake of the 18F-labeled HER2 specific aptamer into the tumor compared to the that of HER2-negative cell tumor(p = 0.033). HER2 aptamer was able to preferentially bind to HER2-positive breast cancer cells both in vitro and in vivo, by recognizing HER2 structure on the surface of these cells. CONCLUSION: The 18F-labeled aptamer enabled appropriate visualization of HER2 expression by human breast cancer cells. The results suggest that a radiolabeled HER2 aptamer could potentially be applied in the development of treatment strategies or in targeted therapy against HER2-positive breast cancer cells.

Therapeutic effects of a liquid bandage prepared with cellulose powders from Styela clava tunics and Broussonetia kazinoki bark: Healing of surgical wounds on the skin of Sprague Dawley rats.

Cellulose in different forms has extensively been applied in biomedical treatments, including scaffolding, tissue engineering and tissue formation. To evaluate the therapeutic effects of a liquid bandage (LB) prepared with cellulose powders from Styela clava tunics (SCT) and Broussonetia kazinoki bark (BSLB) for healing cutaneous wounds, the remedial effects of a low concentration (LoBSLB) and a high concentration (HiBSLB) of BSLB on skin regeneration and toxicity in Sprague Dawley rats. Results indicated that the total area of skin involved in the surgical wound was lower in the BSLB­treated group compared with the Vehicle­treated group at days 4­12, although some variations were observed in the HiBSLB­treated group. In addition, the BSLB­treated group showed significantly enhanced width of the re­epithelialization region and epidermal thickness when compared with the Vehicle­treated group. Furthermore, significant stimulation in the expression level of collagen­1 and the signaling pathway of VEGF after topical application of BSLB was indicated. No liver or kidney toxicities were detected for either doses of BSLB. Overall, the results of the present study suggest that BSLB accelerates the process of wound healing in surgical skin wounds of Sprague Dawley rats through stimulation of re­epithelialization and connective tissue formation, without any accompanying significant toxicity.

Berberine Suppresses Fibronectin Expression through Inhibition of c-Jun Phosphorylation in Breast Cancer Cells.

PURPOSE: The exact mechanism regulating fibronectin (FN) expression in breast cancer cells has not been fully elucidated. In this study, we investigated the pharmacological mechanism of berberine (BBR) with respect to FN expression in triple-negative breast cancer (TNBC) cells. METHODS: The clinical significance of FN mRNA expression was analyzed using the Kaplan-Meier plotter database (http://kmplot.com/breast). FN mRNA and protein expression levels were analyzed by real-time polymerase chain reaction and western blotting, respectively. RESULTS: Using publicly available clinical data, we observed that high FN expression was associated with poor prognosis in patients with breast cancer. FN mRNA and protein expression was increased in TNBC cells compared with non-TNBC cells. As expected, recombinant human FN significantly induced cell spreading and adhesion in MDA-MB231 TNBC cells. We also investigated the regulatory mechanism underlying FN expression. Basal levels of FN mRNA and protein expression were downregulated by a specific activator protein-1 (AP-1) inhibitor, SR11302. Interestingly, FN expression in TNBC cells was dose-dependently decreased by BBR treatment. The level of c-Jun phosphorylation was also decreased by BBR treatment. CONCLUSION: Our findings demonstrate that FN expression is regulated via an AP-1-dependent mechanism, and that BBR suppresses FN expression in TNBC cells through inhibition of AP-1 activity.

Effect of two treatment protocols for ketosis on the resolution, postpartum health, milk yield, and reproductive outcomes of dairy cows.

We determined the effect of ketosis treatment with propylene glycol (PG) or PG plus l-carnitine and methionine (Metabolase®, Fatro, Bologna, Italy) on the resolution, postpartum health, milk yield, and reproductive performances of dairy cows. Blood from 475 Holstein cows was collected weekly until 4 weeks after calving to measure blood ß-hydroxybutyrate (BHBA) concentrations. Cows with blood BHBA concentration ≥1.2 mmol/L were diagnosed with ketosis and were enrolled. One hundred and fifty cows diagnosed with ketosis were randomly assigned to three treatment groups (Day 0): (1) PG (300 g, PO) for 3 days (PG group, n = 50), (2) PG (300 g, PO) plus l-carnitine (1.25 g) plus methionine (5 g, IV) for 3 days (PG + CM group, n = 50), and (3) no treatment (control group, n = 50). On Day 3, blood was collected to evaluate whether the ketosis had resolved. Cows in the PG and PG + CM groups with blood BHBA ≥1.2 mmol/L were retreated for an additional 2 days, and then blood BHBA concentration was evaluated on Days 5 and 10. Blood glucose and haptoglobin concentrations, rumen fill score (RFS), and body condition score (BCS) were measured on Days 0, 3, 5, and 10. Postpartum complications, milk yield during the first 2 months, and reproductive outcomes were evaluated. The probability of resolution from ketosis was higher (P < 0.05) in the PG + CM group than in the control group on Days 3, 5, and 10 (odds ratio: 2.6-6.3). Blood BHBA in the PG + CM group was lower (P < 0.05) than that of the control group on Days 3 and 5, whereas blood glucose in the PG + CM group was higher (P < 0.05) than that of the control group on Days 3 and 5. RFS in the PG and PG + CM groups was higher than that of the control group on Day 10 (P < 0.01), while BCS loss from Day 0-10 in the control group was higher than those of the PG and PG + CM groups (P < 0.05). Milk yields on the 30th and 60th days postpartum were higher in the PG + CM group than the control and PG groups (P < 0.05). Postpartum complications and intervals between calving and first postpartum insemination or pregnancy did not differ among the groups (P > 0.05). In conclusion, treatment of dairy cows with PG plus l-carnitine and methionine improved the chances of resolution of ketosis and increased milk yield, while affecting neither the incidence of postpartum complications nor reproductive performance.

Bisphenol A Affects on the Functional Properties and Proteome of Testicular Germ Cells and Spermatogonial Stem Cells in vitro Culture Model.

The endocrine disruptor bisphenol A (BPA) is well known for its adverse effect on male fertility. Growing evidence suggests that BPA may interact with testicular germ cells and cause infertility as a result of its estrogenic activity. Objective of current in vitro study was to investigate the proliferation, survivability and stemness properties of mouse testicular germ cells exposed to BPA, and to evaluate possible expression of cellular proteome. Our results showed that germ cell viability and proliferation were not affected by low concentrations (0.01, 0.1, 1, and 10 µM) although significant reduction observed at 100 µM BPA. Germ cell self-renewal and differentiation related marker proteins expression found unchanged at those concentrations. When BPA-exposed germ cells were transplanted into recipient testes, we observed fewer colonies at higher concentrations (10 and 100 µM). Additionally, a significant frequency of recombination failure during meiosis was observed in 10 µM BPA-exposed germ cell transplanted recipient. Moreover, experiment on continuous BPA-exposed and 100 µM BPA-recovered germ cells suggested that spermatogonial stem cells are more potential to survive in adverse environment. Finally, scrutinizing differentially expressed cellular proteins resulted from our proteomic analysis, we conclude that BPA exposure might be associated with several health risks and infertility.

Impact of Thermal Decomposition on Thermal Desorption Instruments: Advantage of Thermogram Analysis for Quantifying Volatility Distributions of Organic Species.

We present results from a high-resolution chemical ionization time-of-flight mass spectrometer (HRToF-CIMS), operated with two different thermal desorption inlets, designed to characterize the gas and aerosol composition. Data from two field campaigns at forested sites are shown. Particle volatility distributions are estimated using three different methods: thermograms, elemental formulas, and measured partitioning. Thermogram-based results are consistent with those from an aerosol mass spectrometer (AMS) with a thermal denuder, implying that thermal desorption is reproducible across very different experimental setups. Estimated volatilities from the detected elemental formulas are much higher than from thermograms since many of the detected species are thermal decomposition products rather than actual SOA molecules. We show that up to 65% of citric acid decomposes substantially in the FIGAERO-CIMS, with ∼20% of its mass detected as gas-phase CO2, CO, and H2O. Once thermal decomposition effects on the detected formulas are taken into account, formula-derived volatilities can be reconciled with the thermogram method. The volatility distribution estimated from partitioning measurements is very narrow, likely due to signal-to-noise limits in the measurements. Our findings indicate that many commonly used thermal desorption methods might lead to inaccurate results when estimating volatilities from observed ion formulas found in SOA. The volatility distributions from the thermogram method are likely the closest to the real distributions.

Enrichment and In Vitro Culture of Spermatogonial Stem Cells from Pre-Pubertal Monkey Testes.

Spermatogonial stem cells (SSCs) are essential for spermatogenesis throughout the lifespan of the male. However, the rarity of SSCs has raised the need for an efficient selection method, but little is known about culture conditions that stimulate monkey SSC proliferation in vitro. In this study, we report the development of effective enrichment techniques and in vitro culturing of germ cells from pre-pubertal monkey testes. Testis cells were analyzed by fluorescence-activated cell sorting techniques and were transplanted into the testes of nude mice to characterize SSCs. Thy-1-positive cells showed a higher number of colonies than the unselected control after xenotransplantation. Extensive colonization of monkey cells in the mouse testes indicated the presence of highly enriched populations of SSCs in the Thy-1-positive sorted cells. Furthermore, monkey testis cells were enriched by differential plating using extracellular matrix, laminin, and gelatin, and then cultured under various conditions. Isolation of monkey testicular germ cells by differential plating increased germ cell purity by 2.7-fold, following the combinational isolation method using gelatin and laminin. These enriched germ cells actively proliferated under culture conditions involving StemPro medium supplemented with bFGF, GDNF, LIF, and EGF at 37 °C. These results suggest that the enrichment and in vitro culture method proposed in the present study for harvesting a large number of functionally active monkey SSCs can be applied as the basis for efficient in vitro expansion of human SSCs.

Withdrawal: Specific nephrotoxicity and cardiotoxicity of BT-CAL®, Sigma Anti-bonding Molecule Calcium Carbonate, in mice.

[This retracts the article on p. 7 in vol. 29, PMID: 23573102.].

Hybridization-based aptamer labeling using complementary oligonucleotide platform for PET and optical imaging.

Aptamers are promising next-generation ligands used in molecular imaging and theragnosis. Aptamers are synthetic nucleic acids that can be held together with complementary sequences by base-pair hybridization. In this study, the complementary oligonucleotide (cODN) hybridization-based aptamer conjugation platform was developed to use aptamers as the molecular imaging agent. The cODN was pre-labeled with fluorescent dye or radioisotope and hybridized with a matched sequence containing aptamers in aqueous conditions. The cODN platform-hybridized aptamers exhibited good serum stability and specific binding affinity towards target cancer cells both in vitro and in vivo. These results suggest that the newly designed aptamer conjugation platform offers great potential for the versatile application of aptamers as molecular imaging agents.

Effects of paracrine factors on CD24 expression and neural differentiation of male germline stem cells.

Spermatogonial stem cells (SSCs) are adult male germ cells that develop after birth. Throughout the lifetime of an organism, SSCs sustain spermatogenesis through self-renewal and produce daughter cells that differentiate into spermatozoa. Several studies have demonstrated that SSCs can acquire pluripotency under appropriate culture conditions, thus becoming multipotent germline stem cells (mGSCs) that express markers of pluripotency in culture and form teratomas following transplantation into immunodeficient mice. In the present study, we generated neural precursor cells expressing CD24, a neural precursor marker, from pluripotent stem cell lines and demonstrated that these cells effectively differentiated along a neural lineage in vitro. In addition, we found that paracrine factors promoted CD24 expression during the neural differentiation of mGSCs. Our results indicated that the expression of CD24, enhanced by a combination of retinoic acid (RA), noggin and fibroblast growth factor 8 (FGF8) under serum-free conditions promoted neural precursor differentiation. Using a simple cell sorting method, we were able to collect neural precursor cells with the potential to differentiate from mGSCs into mature neurons and astrocytes in vitro.