Identification of a novel histone H2A mono-ubiquitination-inhibiting cell-active small molecule.

Histone H2A mono-ubiquitination plays important roles in epigenetic gene expression and is also involved in tumorigenesis. Small molecules controlling H2A ubiquitination are of interest as potential chemical tools and anticancer drugs. To identify novel small molecule inhibitors of H2A ubiquitination, we synthesized and evaluated several compounds designed based on PRT4165 (1), which is a reported histone ubiquitin ligase RING1A inhibitor. We found that compound 11b strongly inhibited the viability and reduced histone H2A mono-ubiquitination in human osteosarcoma U2OS cells. Therefore, compound 11b is a promising lead compound for the development of H2A histone ubiquitination-inhibiting small molecules.

Correction: Electronic Health Record-Nested Reminders for Serum Lithium Level Monitoring in Patients With Mood Disorder: Randomized Controlled Trial.

[This corrects the article DOI: 10.2196/40595.].

Electronic Health Record-Nested Reminders for Serum Lithium Level Monitoring in Patients With Mood Disorder: Randomized Controlled Trial.

BACKGROUND: Clinical guidelines recommend regular serum lithium monitoring every 3 to 6 months. However, in the real world, only a minority of patients receive adequate monitoring. OBJECTIVE: This study aims to examine whether the use of the electronic health record (EHR)-nested reminder system for serum lithium monitoring can help achieve serum lithium concentrations within the therapeutic range for patients on lithium maintenance therapy. METHODS: We conducted an unblinded, single-center, EHR-nested, parallel-group, superiority randomized controlled trial comparing EHR-nested reminders with usual care in adult patients receiving lithium maintenance therapy for mood disorders. The primary outcome was the achievement of therapeutically appropriate serum lithium levels between 0.4 and 1.0 mEq/L at 18 months after enrollment. The key secondary outcomes are included as follows: the number of serum lithium level monitoring except for the first and final monitoring; exacerbation of the mood disorder during the study period, defined by hospitalization, increase in lithium dose, addition of antipsychotic drugs or mood stabilizers, or addition or increase of antidepressants; adherence defined by the proportion of days covered by lithium carbonate prescription during the study period. RESULTS: A total of 111 patients were enrolled in this study. A total of 56 patients were assigned to the reminder group, and 55 patients were assigned to the usual care group. At the follow-up, 38 (69.1%) patients in the reminder group and 33 (60.0%) patients in the usual care group achieved the primary outcome (odds ratio 2.14, 95% CI 0.82-5.58, P=.12). The median number of serum lithium monitoring was 2 in the reminder group and 0 in the usual care group (rate ratio 3.62; 95% CI 2.47-5.29, P<.001). The exacerbation of mood disorders occurred in 17 (31.5%) patients in the reminder group and in 16 (34.8%) patients in the usual care group (odds ratio 0.97, 95% CI 0.42-2.28, P=.95). CONCLUSIONS: We found insufficient evidence for an EHR-nested reminder to increase the achievement of therapeutic serum lithium concentrations. However, the number of monitoring increased with relatively simple and inexpensive intervention. The EHR-based reminders may be useful to improve quality of care for patients on lithium maintenance therapy, and they have potentials to be applied to other problems. TRIAL REGISTRATION: University Hospital Medical Information Network Clinical Trials Registry UMIN000033633; https://tinyurl.com/5n7wtyav.

Comparison of the Efficacy of Sub-Tenon versus Intravitreal Triamcinolone Acetonide Injection during Cataract Surgery for Diabetic Macular Edema.

PURPOSE: We compared the efficacy of sub-Tenon triamcinolone acetonide (STTA) to intravitreal triamcinolone aceto-nide (IVTA) injections during cataract surgery (CS) for patients with diabetic macular edema (DME). METHODS: The medical records of 33 eyes (26 patients) with DME which had undergone CS with STTA were compared to those of 34 eyes (27 patients) with DME which had undergone CS with IVTA. Central foveal thickness and best-corrected visual acuity (BCVA) were measured at the baseline and 1, 3, and 6 months after the surgery. RESULTS: The BCVAs after STTA and IVTA were significantly improved at 3 and 6 months. Thirteen eyes in the IVTA group and 21 eyes in the STTA group required other therapies (p < 0.05). One case developed intraocular pressure elevation after IVTA and underwent selective la ser trabeculoplasty. CONCLUSIONS: Ophthalmologists should consider the merits and demerits of IVTA and STTA for DME treatment after CS.

Cryoprotective effect of antifreeze polyamino-acid (Carboxylated Poly-l-Lysine) on bovine sperm: A technical note.

In this study, investigating Carboxylated Poly-l-Lysine (CPLL) for its effectiveness as a new cryoprotectant for bovine sperm is aimed. CPLL is an ampholytic polymer compound, has cryoprotective properties similar to those of anti-freeze protein. The cryopreservation medium used for control group consisted of 6.5% (v/v) glycerol, the cryopreservation medium used for experimental group consisted of 3.25% (v/v) glycerol + 0.5% (w/v) CPLL. There was no consequential difference in sperm motility parameter after thawing whereas there was huge distinction for sperm membrane integrity rate (control vs experimental; 49.6 vs 60.7%, P < 0.01). Conception rate of artificial insemination of experimental group was significantly higher than that of control group (79.0% vs 53.1%, P < 0.01). These results suggest CPLL has protected sperm membrane and leads to improve fertility. This is the first report using CPLL for bovine sperm cryopreservation, it is also expected CPLL can be applied to other animal species.

Cryopreservation of bovine somatic cells using antifreeze polyamino-acid (carboxylated poly-L-lysine).

Carboxylated poly-L-lysine (CPLL) is an ampholytic polymer compound, obtained by converting 65 mol% of amino groups to carboxyl groups after synthesizing ε-poly-l-lysine aqueous solution and succinic anhydride. CPLL has cryoprotective properties similar to those of anti-freeze protein. The addition of CPLL to freezing medium has been reported to improve the post-thawing survival rate of murine cells, human induced pluripotent stem (iPS) cells, embryonic stem (ES) cells and embryos. In this study, investigating CPLL for its effectiveness as a new cryoprotective material is aimed. In experiments with bovine somatic cells, CPLL was suggested to have an equal or superior cryoprotective effect to dimethyl sulfoxide (DMSO), the conventional material for cellular frozen storage, based on the results for post-thawing cell survival and proliferation rates. CPLL was demonstrated to have another advantage; thawed cells can be cultured without removing the cryopreservation medium when CPLL is used, but not when DMSO is used. These results suggest that CPLL could be used as cryoprotective material for bovine cells. It is also expected that CPLL can be applied to embryo and oocytes storage for cattle, and similar functions for cells and embryos of other animal species.

Effect of probiotics on blood fatty acid metabolism during the late middle stage of fattening period in Japanese Black cattle.

This study was conducted to investigate the effects of probiotics administration on fatty acid metabolism in Japanese Black cattle as per changes in blood fatty acid concentrations and blood biochemical tests. Eighteen clinically healthy Japanese Black female fattening cattle bred on the same fattening farm were randomly classified into the probiotics administration group (n=9) or the control group (n=9). In the probiotics administration group, 50 g of probiotics were started per animal per day at the age of 18 months, and the administration period was 2 months from the start date of the study. Blood was collected twice before starting the probiotics administration and at 2 months after starting the probiotics administration. In the probiotics administration group, palmitic, linoleic, arachidonic and α-linolenic acid tended to be higher at the end of the administration compared with those before probiotics administration. Additionally, as a result of multiple comparison test, monounsaturated fatty acids at Post was significantly higher, and the ω6 / ω3 ratio was significantly lower than in the control group. Vitamin A, E and albumin were significantly higher at the end of the administration than in the control group. In this study that administering probiotics to Japanese Black cattle in the late middle stage of fattening period did not have a significant effect on fatty acid metabolism during feed digestion and absorption, but suggested that may alter some blood fatty acids concentrations.

A clinical case of a subepiglottic cyst in a Japanese Black calf.

A 36-day-old Japanese Black calf exhibited wheezing associated with dyspnea from birth. Arterial blood gas analysis revealed a low oxygen partial pressure of 51 mmHg, low oxygen saturation of 83%, and high carbon dioxide partial pressure of 58.8 mmHg. Computed tomography, endoscopy, and ultrasonography showed cyst formation under the epiglottis. When the cyst was aspirated under ultrasonic guidance to secure the airway, 30 ml of viscous white turbid content was aspirated. The cyst shrank immediately after aspiration, but the wheezing and respiratory symptoms resumed 7 days after aspiration. Therefore, the cyst was surgically removed from the ventral side of the neck. No cyst remodeling was observed 30 days after surgical removal.

Utility of carboxylated poly L-lysine for the liquid storage of bottlenose dolphin semen.

Assisted reproduction techniques are required to maintain a genetically diverse captive population of bottlenose dolphins. These techniques include semen preservation, and liquid storage has been proposed as a suitable alternative to cryopreservation, but the optimum conditions, in terms of temperature, duration, and media, have yet to be fully established. The aim of this study, therefore, was to determine the optimum temperature for the liquid storage of dolphin semen during a 14-day period and the usefulness of carboxylated poly-L-lysine (CPLL) as an additive to the semen extender used for the liquid storage. The semen was collected from a mature male dolphin housed at the Kagoshima Aquarium, Japan, transferred into a Beltsville (BF5F) extender, and analyzed for motility and characteristics after five-fold dilution. The optimum temperature was determined by evaluating sperm viability after liquid storage at 4, 17, or 36 °C, and the usefulness of CPLL was evaluated at concentrations of 0%, 0.5%, 1.0%, 2.0%, and 3.0% (v/v) at the optimum temperature. Sperm stored at 4 â„ƒ had a greater motility maintenance compared with samples stored at 17 or 36 â„ƒ. The most efficacious storage regimen at various time points occurred when there was addition of CPLL at 1.0% (v/v) in terms of sperm motility and other relevant determinations, with this storage approach having greater efficacy that samples stored without CPLL. The most efficacious processes for preserving bottlenose dolphin sperm functions is storage at 4 °C and with there being semen extender supplementation of 1% CPLL.

Alterations of growth, blood biochemical components and hormone profiles by intensified nutrition in growth retarded Japanese Black cattle.

In order to determine the clinical conditions of Japanese Black (JB) cattle with growth retardation, we determined the changes of body growth, blood profiles of metabolism and hormones caused by intensified nutrition (sufficient total digestible nutrients and digestible crude protein for a target daily gain set at 1.2-1.3 kg/day) in three cattle. The daily gain (DG) was increased during the intensified period (Intense) compared with the preparation period (Pre), but the DG in the Intense period was 36-66% of the target DG. Serum albumin, total cholesterol, insulin and IGF-1 increased during the Intense period compared with the Pre period. Serum GH showed high levels in the Pre period, whereas it showed lower levels in the Intense period. These results suggested that the present growth retarded cattle had abnormalities in their metabolic systems and lacked nutrient absorption.

Bovine milk transcriptome analysis reveals microRNAs and RNU2 involved in mastitis.

Mastitis is a common inflammatory infectious disease in dairy cows. To understand the microRNA (miRNA) expression profile changes during bovine mastitis, we undertook a genome-wide miRNA study of normal milk and milk that tested positive on the California mastitis test for bovine mastitis (CMT+). Twenty-five miRNAs were differentially expressed (23 miRNAs upregulated and two downregulated) during bovine mastitis relative to their expression in normal milk. Upregulated mature miR-1246 probably derived from a U2 small nuclear RNA rather than an miR-1246 precursor. The significantly upregulated miRNA precursors and RNU2 were significantly enriched on bovine chromosome 19, which is homologous to human chromosome 17. A gene ontology analysis of the putative mRNA targets of the significantly upregulated miRNAs showed that these miRNAs were involved in binding target mRNA transcripts and regulating target gene expression, and a Kyoto Encyclopedia of Genes and Genomes pathway analysis showed that the upregulated miRNAs were predominantly related to cancer and immune system pathways. Three novel miRNAs were associated with bovine mastitis and were relatively highly expressed in milk. We confirmed that one of the novel mastitis-related miRNAs was significantly upregulated using a digital PCR system. The differentially expressed miRNAs were involved in human cancers, infections, and immune-related diseases. The genome-wide analysis of miRNA profiles in this study provides insight into bovine mastitis and inflammatory diseases. DATABASES: The miRNAseq generated for this study can be found in the Sequence Read Archive (SRA) under BioProject Number PRJNA421075 and SRA Study Number SRP126134 (https://www.ncbi.nlm.nih.gov/bioproject/PRJNA421075).

Age-related alterations in peripheral leukocyte population of healthy Holstein dairy cows during the pre-calving period.

Older cows show a high incidence of infectious diseases during the periparturient period. The periparturient infectious diseases are closely associated with the immune function of dairy cows during the pre-calving period. In order to evaluate the relationship between the immune cell population and age in the cows during the pre-calving period, we obtained blood samples from 170 dairy cows during the pre-calving period. We chose only healthy cows, which did not develop clinical diseases within 2 weeks after the calving in this study. The animals were divided into 4 groups based on their parity: in their 1st pregnancy (Group 1), in their 2nd pregnancy (Group 2), in their 3rd calving (Group 3) and in more than 3rd pregnancy (Group 4). The numbers of the peripheral blood CD3(+)TcR1-N12(+) and MHC class-II(+)CD14(-) lymphocytes were significantly higher in Group 1 compared to Group 4. This result indicated that the lower gammadeltaT cells and B cells in older cows compared with heifer during pre-calving period.

Relationship between immune function and serum vitamin A in Japanese black beef cattle.

This study evaluated the relationship between the serum vitamin A level and immune function in seventy non-diseased Japanese Black (JB) cattle during their fattening stages. The animals were divided into two groups, a Low Vitamin A (VA) group (N=9) with below 30 IU/dl of serum VA and a Control group (N=61). Blood samples were collected for biochemical analysis and examination of the leukocyte population and cytokine mRNA expression. The numbers of CD3(+)WC1(+) T cells and MHC class-ll(+)CD14(-) B cells were significantly lower in the Low VA group than in the Control group (P<0.05). The IFN-gamma/IL-4 rate was significantly lower in the Low VA group, while IL-4 was higher and IFN-gamma was lower in the Low group compared with the Control group. This study indicated that immune function imbalance was present in JB cattle with low serum VA levels during the fattening stage.

Effect of lidocaine hydrochloride on the function of bovine peripheral leukocytes.

To clarify the effect of lidocaine hydrochloride (Lid) on bovine peripheral granulocyte phagocytosis, adhesion molecule expression of leukocytes and peripheral blood mononuclear cell mRNA expression of cytokines were investigated. Lid was added to blood samples at a final concentration of 0 (only PBS; Cont), 0.2 mg/ml or 2.0 mg/ml. Phagocytosis of granulocytes was significantly decreased by addition of 2.0 mg/ml of Lid. CD18 expression of granulocytes and mononuclear cells were significantly reduced by addition of 2.0 mg/ml of Lid. IL-1beta and IL-8 mRNA expressions of mononuclear cells were also significantly reduced by addition of 2.0 mg/ml of Lid other hand. These results suggest that Lid might reduce the protective immunity of cows. On the other hand, reduction of CD18, IL-1beta and IL-8 mRNA expression also indicates that Lid has an anti-inflammatory effect in cows.

Peripheral blood leukocyte subpopulation of dairy cows with digital dermatitis and effect of hoof trimming with antibiotic treatment.

In the present study, 30 cows were used to evaluate the changes in the peripheral blood leukocyte subpopulation of dairy cows with digital dermatitis (DD) following hoof trimming and antibiotic treatment. The cows were divided into two groups; 18 cows (DD group) had DD on both hind feet, and 12 cows (control group) had four feet with no clinical abnormalities. The DD group was further divided into two groups based on the treatment; the antibiotic group (8 cows) was treated with only 2% lincomycin liquid spray once daily for 3 days, and the trimmed group (10 cows) received trimming of hooves as well as treatment with 2% lincomycin liquid spray. The plasma cortisol concentration was significantly higher in both DD groups before treatment than in the control group, and it decreased significantly after hoof trimming in the trimmed group. The number of CD3(+), CD4(+), WC1(+) and CD21(+) cells in both DD groups before treatment was significantly lower than that of the control group. The number of CD3(+), CD4(+), WC1(+) and CD21(+) cells in the trimmed group increased after treatment. These results indicated that cows with DD suffer from stress and reduced number of T and B cells. Treatment of DD with both hoof trimming and 2% lincomycin liquid spray was effective for reducing the stress and bringing the immune cell number back to the normal range.

Effect of body condition score of the dairy cow on the in vitro immune response of peripheral blood mononuclear cells to progesterone stimulation.

Immune function in cows is closely associated with their physical and hormonal conditions. In order to clarify the relationship between the body condition score (BCS) of lactating dairy cows and the immune response to progesterone (P(4)) in vitro, we examined whether lower BCS in dairy cows affects the responsiveness of peripheral blood mononuclear cells (PBMCs) to P(4) added in to culture medium. Forty-two non-pregnant healthy Holstein dairy cows were examined at 61 to 120 days after calving. The cows were divided into the following two groups; Low BCS group (N=20), which had a BCS of less than 2.25, and a Control group (N=22), which had a BCS over 2.75. PBMCs were stimulated with P(4) (1 microg/mL) and/or phytohemagglutinin (PHA), and the levels of cytokine mRNA were analyzed. In the Low BCS group, a significantly lower IFN-gamma level was stimulated by PHA only compared with the Control group. The combination of P(4) and PHA significant decreased the IFN-gamma/IL-4 ratio in the Control group, but this reaction was not found in the Low BCS group. Our data indicated that expression of IFN-gamma mRNA was basically lower in the low BCS dairy cows and that addition of P(4) did not suppress the cellular immune function in these cows. In this study, we observed that P(4) reduced the cellular immune response in the adequate BCS cows, whereas immunosuppression by P(4) was not found in the PBMCs of the low BCS cows, which already had a lower level of immune function.

Effect of combined vaccination for Pasteurella multocida, Mannheimia haemolytica, and Histophilus somni to prevent respiratory diseases in young Japanese Black calves in the field.

To evaluate the preventive effects of combined vaccination for Pasteurella multocida, Mannheimia haemolytica and Histophilus somni on respiratory diseases in Japanese Black calves, 295 calves at one farm were alternately assigned to two groups; 147 calves received the vaccine at 4 and 8 weeks of age (vaccination group), and the other 148 calves did not receive vaccine (control group). The incidences of respiratory diseases were 25.9 and 70.9% in the vaccination and control groups, respectively, and the odds ratio for comparison between the two groups was 0.143 (95% confidence interval: 0.086-0.238). Administration of the multiple vaccine to Japanese black calves might be one of effective factor for prevention of respiratory diseases.

Effect of hoof trimming before the dry period on productive performance in perinatal dairy cows.

To investigate the effect of hoof trimming before the dry period, the hooves of 10 cows (trimmed group) were trimmed at 79.6 +/- 8.6 days before parturition and the hooves of 52 cows were left untrimmed (control group). Blood biochemistry and hormone concentrations were investigated for 6 cows from each group. The daily milk yield after parturition in the trimmed group tended to be higher than that of the control group. Between 0 and 30 days after parturition, the levels of non-esterified fatty acids were significantly lower stet, and the plasma glucose and glucose disposal rates were significantly higher in the trimmed group. The plasma cortisol concentration was stable before and after parturition in the trimmed group. Hoof trimming before the dry period appears to reduce stress and maintain the nutritional conditions of perinatal dairy cows.

Effects of active egg white product/ Clostridium butyricum Miyairi 588 additive on peripheral leukocyte populations in periparturient dairy cows.

The leukocyte populations of periparturient dairy cows were analyzed after administration of active egg white/Clostridium butyricum Miyairi additive. Sixty-eight Holstein milking cows were divided into 3 groups. Group A was administered active egg white product (AEWP)/Clostridium butyricum Miyairi 588 (Miyairi 588) additive (n=23). Group B was administered Miyairi 588 only (n=23), and Group C was the control group (n=22). The challenged groups were administered 100 g of AEWP + Miyairi 588, or Miyairi 588 alone, daily for 60 days from 1 month before until 1 month after paturition. Blood samples were collected from all groups three times (1 month before, 1 week after and 1 month after parturition) for analysis of the peripheral leukocyte population. The results showed significantly higher numbers of CD4+ cells in Group A compared with Group C 1 week after paturition. AEWP/Miyairi 588 additive may enhance the number of CD4+ T cells in periparturient dairy cows.

Changes in peripheral blood mononuclear cells after calving in lactating cows with serious decrease of body weight before and after calving.

The aim of this study was to clarify the hormonal and immunological changes in periparturient dairy cows with excessive weight loss following calving. We analyzed leukocyte populations and the peripheral blood mononuclear cell (PBMC) mRNA of IFN-gamma, IL-4, growth hormone (GH)-receptor (R) and insulin -R and measured the GH and insulin concentrations. Ten dairy cows fed in one herd were used in this study and were divided into two groups. Five cows that experienced loss of body weight following calving of over 10% comprised the Weight Loss Group, and the remaining five cows, which experienced loss of body weight of 5% or less following calving, served as the Control Group. The Weight Loss Group had high serum cholesterol and low blood urea nitrogen levels compared with the Control Group throughout the period of observation. In regard to the leukocyte populations, there were significantly fewer CD4(+)CD45R-T cells in the Weight Loss Group 1 to 2 months after calving compared with the Control Group. The percentage of IFN-gamma/IL-4 among PBMCs in the Weight Loss Group tended to be lower than in the Control Group throughout the observation period. In the Weight Loss Group, the levels of GH-R during month 2 were lower than in the Control Group. We concluded that excessive loss of body weight in dairy cows following calving is associated with depressed cellular immune function.