A systematic review of the behaviours associated with depression in people with severe-profound intellectual disability.

The assessment of depression in people with severe to profound intellectual disability (severe-profound ID) is challenging, primarily due to inability to report internal states such as mood, feelings of worthlessness and suicidal ideation. This group also commonly presents with challenging behaviours (e.g. aggression and self-injury) with debate about whether these behaviours should be considered 'depressive equivalents' or are sensitive for, but not specific to, depression in severe-profound ID. We conducted a systematic review exploring behaviours associated with depression and low mood in individuals with severe-profound ID. The review was conducted in accordance with Preferred Reporting Items for Systematic Reviews and Meta-Analyses (2009) guidelines. Three electronic databases were searched (Embase, PsycINFO and Ovid MEDLINE), and 13 studies were included and rated for quality. Few studies were rated as having high methodological quality. Behaviours captured by standard diagnostic schemes for depression (e.g. Diagnostic and Statistical Manual of Mental Disorders and International Classification of Diseases) showed a relationship with depression in severe-profound ID, including the two core symptoms (depressed affect and anhedonia), as well as irritability, sleep disturbance, psychomotor agitation, reduced appetite and fatigue. Challenging behaviours such as aggression, self-injury, temper tantrums, screaming and disruptive behaviour were associated with depression. Challenging behaviours show a robust relationship with depression. Whilst these behaviours may suggest an underlying depression, study limitations warrant caution in labelling them as 'depressive equivalents'. These limitations include not controlling for potential confounds (autism, other affective disorders and pain) and bias associated with comparing depressed/non-depressed groups on the same behavioural criteria used to initially diagnose and separate these groups. Future studies that use depressive measures designed for ID populations, which control for confounds and which explore low mood irrespective of psychiatric diagnosis, are warranted to better delineate the behaviours associated with depression in this population (PROSPERO 2018: CRD42018103244).

A rare disease with a rare presentation: hemi-atrophy caused by fibromuscular dysplasia in a 27 month old girl.

We discuss the case of a 27-month-old girl afflicted with fibromuscular dysplasia. She presented with hemiatrophy of left upper and lower limbs, nail dystrophy, ulcers on the tips of her toes, cold and painful limbs, foot drop, and hypertension. The initial appearance started at 2 months of age and other diagnoses such as complex regional pain syndrome, reflex sympathetic syndrome, vasculitis and coagulation disorders had been considered. Angiography revealed that all the arterial branches of the left lower and upper limbs, from brachial to ulnar and radial, and from iliac and femoral to tibialis arteries were affected. Sural nerve biopsy confirmed the diagnosis. In the follow-up visits until 2 years after the patient's discharge she did not develop any new problem and her blood pressure was controlled by enalapril and amlodipine.

Melatonin as an anti-inflammatory agent in radiotherapy.

Radiotherapy is one of the most relevant treatment options for cancer therapy with or without other treatment modalities including immunotherapy, surgery and chemotherapy. Exposure to heavy doses of ionizing radiation during radiotherapy results in short and long term side effects. It appears that many of these side effects are linked to inflammatory responses during treatment or after prolonged use. Inflammation is mediated by various genes and cytokines related to immune system responses caused by massive cell death following radiotherapy. This phenomenon is more obvious, particularly after exposure to clinical doses of radiotherapy. Inflammation is involved in the amplification of acute responses, genomic instability and also long term pathological changes in normal tissues. Moreover, inflammation attenuates responses of the tumor to radiotherapy through some mechanisms such as angiogenesis. Thus, the management of inflammation is one of the most interesting aims in cancer radiotherapy. Melatonin, known as a natural product in the body, has been of much interest for its anti-inflammatory properties. Some studies have proposed melatonin as a novel anti-inflammation agent. This literature review will concentrate on the anti-inflammatory properties of melatonin that may help the management of different inflammatory signaling pathways in both tumor and normal tissues.

The Anti-apoptotic Mechanism of Metformin Against Apoptosis Induced by Ionizing Radiation in Human Peripheral Blood Mononuclear Cells.

BACKGROUND: In a previous article, we showed that metformin (MET) can reduce ionizing radiation (IR) induced apoptosis in human peripheral blood mononuclear cells. However, the anti-apoptotic mechanism of MET against IR remains unclear. The present study attempts to investigate the mechanism of action of MET in limiting X-ray induced apoptosis in human peripheral blood mononuclear cells. MATERIAL AND METHODS: Mononuclear cells were treated with MET for 2 hours and irradiated with 6 MV X-rays. The gene expression levels of BAX, CASP3 and BCL2 were determined 24 hours post irradiation using real time quantitative polymerase chain reaction (qPCR) technique. Furthermore, the protein levels of BAX, CASP3 and BCL2 were analyzed by Western blotting assay. RESULTS: Radiation exposure increased the expressions of BAX and CASP3 genes, and decreased the expression of BCL2 gene in mononuclear cells. Conversely, an increase in BCL2 gene expression along with a decrease in BAX and CASP3 genes expression was observed in MET plus irradiated mononuclear cells. It was found that radiation increased BAX/BCL2 ratio, while MET pretreatment reduced these ratios. Also, treatment with MET without irradiation did not change the expressions of BAX, CASP3 and BCL2 genes. On the other hand, downregulated expression of BCL2 protein and upregulated expressions of BAX and CASP3 proteins were found in 2 Gy irradiated mononuclear cells, while pretreatment with MET significantly reversed this tendency. CONCLUSION: These results suggest that MET can protect mononuclear cells against apoptosis induced by IR through induction of cellular anti-apoptotic signaling.Key words: ionizing radiation - metformin - apoptosis - genes - proteins - blood cells.

Association of interleukin-16 polymorphisms with disease progression and susceptibility in endometriosis.

Interleukin-16 (IL-16) is a multifunctional pro-inflammatory cytokine that was previously found in association with complex disorders, and it is now cleared that this cytokine plays a critical role in regulation of cellular functions such as homoeostasis. Due to the complexity of endometriosis and its resemblance to cancer, we designed present case-control study to determine the effects of genetic polymorphisms of the human IL-16 gene on Iranian women's susceptibility to endometriosis. A total of 126 patients with endometriosis (stages I-IV) and 144 healthy women as control group were recruited to the study. We genotyped four single nucleotide polymorphisms of IL-16 gene (rs11556218 T>G, rs4778889 T>C, rs4072111 C>T and rs1131445 C>T). Genotyping was performed using PCR and restriction fragment length polymorphism. Our results showed that genotype distribution in two exonic polymorphisms including rs11556218 and rs4072111 was significantly different between Endometriosis patients and healthy individuals (P < 0.05). We have also found an association between rs4072111 and rs1131445 with progression to the severe stages (III-IV) of endometriosis (P < 0.05). Finally, we may conclude that IL-16 gene polymorphisms are highly associated with increased risk of endometriosis and could be considered as a susceptibility factor for endometriosis.

Expression of bovine Ecat1 gene in immature and in vitro matured oocytes as well as during early embryonic development.

Ecat1 is a maternal effect gene that is exclusively expressed in oocytes and embryonic stem cells, and has an important role in pre-implantation development. This study was designed to investigate the expression of bovine Ecat1 gene in immature and in vitro matured oocytes as well as during early embryonic development, and also Ecat1 protein localization. Samples were obtained from slaughtered animals. RNA extractions were carried out from ovary, immature and in vitro matured oocytes and also different stages of embryonic development (2-, 4-, 8- to 16-cell stages and blastocysts). RT-PCR analysis revealed the expression of Ecat1 in ovary, oocytes and embryos. Analysis in FGENESH online tool predicted three exons and one transcription start site (TSS) in Ecat1 gene, and the 3' RACE-PCR result showed that just one splice variant was amplified. By quantitative real-time PCR technique, we showed that Ecat1 transcript increased at 8- to 16-cell-stage embryos and decreased in blastocyst stage (p < 0.05). Immunofluorescence analysis showed cytoplasmic localization of Ecat1 protein in bovine oocytes. Results demonstrated bovine Ecat1 expression at protein level and also indicated that Ecat1 has a significant higher embryonic expression at 8- to 16-cell stage. This embryonic expression is probably required for further developmental stages.

Antioxidant effects of clove bud (Syzygium aromaticum) extract used with different extenders on ram spermatozoa during cryopreservation.

Clove bud (Syzygium aromaticum) extract was added at concentrations of 0, 35, 75, and 115 µg/ml to ovine semen extenders in order to investigate the antioxidant activities of clove bud extract and its effects on semen quality parameters after cryopreservation of ram spermatozoa. The basic extender was composed of Tris, egg yolk, and glycerol. Two other extenders were prepared by substitution of egg yolk with either LDL or egg yolk+SDS. The DPPH inhibition test was employed to assess the antioxidant activity of clove bud extract. Results showed that, compared to vitamin E, clove bud extract had a higher antioxidant activity. Better sperm motility and movement characteristics (P<0.05) were observed in the semen diluted with medium containing egg yolk+SDS than in that containing egg yolk and LDL. Progressive motility and movement characteristics of the sperm were significantly improved (P<0.05) by adding 35 and/or 75 µg/ml of clove bud extract to semen extenders. Sperm viability and plasma membrane integrity were also higher (P<0.05) in the semen exposed to medium containing egg yolk+SDS and 75 µg of clove buds extract after cryopreservation processes. Higher levels of clove bud extract, however, had adverse effects on all the sperm quality parameters and significantly reduced (P<0.05) the motility, movement parameters, viability, and plasma membrane integrity of ovine sperm. It was concluded that the clove bud extract had an antioxidant potential that makes it useful for addition to semen extenders and that the best results are obtained with a maximum clove bud extract of 75 µg/ml. Moreover, the combination of egg yolk and a detergent was found to improve sperm quality after the cooling and freeze-thawing processes.

Comparative evaluation of cervical pulpotomy and pulpectomy for primary molars with irreversible pulpitis: a multicentre randomised controlled trial.

PURPOSE: The objective of this multicentre randomised controlled trial was to compare the clinical/radiographic outcomes of cervical pulpotomy using calcium-enriched mixture cement (PCEM) and pulpectomy using Metapex (PM) in primary molars with irreversible pulpitis (IP). METHODS: A total of 134 primary molars from 94 children were randomly assigned to two intervention groups: the PCEM group (n = 74) and the PM group (n = 60). Baseline characteristics including age/gender/molar type/tooth type/jaw were recorded. The primary outcome measures were clinical/radiographic success rates assessed at the first and second follow-up appointments. Secondary outcomes included reasons for clinical/radiographic failures. Multiple logistic regression analysis was performed to determine the impact of various factors on the success rates. RESULTS: The mean age of the participants in both groups was similar (PCEM group: 5.4 years, PM group: 5.5 years). Gender distribution, molar type, tooth type, jaw, and number of practitioners were comparable between the groups. The clinical success rate at the first follow-up was 98.6% in the PCEM group and 96.4% in the PM group. At the second follow-up, the clinical success rate was 97.1% in the PCEM group and 91.1% in the PM group. The radiographic success rates at the first and second follow-up were 98.6% and 96.4% in the PCEM group and 96.4% and 91.1% in the PM group, respectively. Multiple logistic regression analysis did not reveal any significant association between the success rates and age/gender/molar type/jaw, or treatment groups (P > 0.05). CONCLUSION: In primary molars with IP, both simple/conservative cervical pulpotomy using calcium-enriched mixture cement and pulpectomy using Metapex demonstrated high clinical/radiographic success rates. No significant differences were observed between the two treatment modalities. These findings suggest that both techniques can be considered effective treatment options for managing primary molars with IP. TRIAL REGISTRATION NUMBER: Trial registration number: IRCT20201226049838N1, retrospectively registered on 12 January 2021.

National guidelines for outbreak investigation: an evaluation study.

This paper reports an independent epidemiological study to evaluate the validity of the results of an official investigation into an outbreak of gastroenteritis at a university campus in Yasuj, central-south Islamic Republic of Iran. The official report of the outbreak by the Department for Disease Control at the provincial health centre found only 65 cases over a 5-day period, all females, living in the student halls of residence. This contrasts with a questionnaire survey of 963 students at the same university, which found 395 students (192 males and 203 females), living in residences and at home, who reported at least 1 gastrointestinal symptom over a 12-week period. Within this period at least 2 outbreaks occurred. Such a large discrepancy between the official report and the current study suggests that the health services and the public may have been misled about the proper response to the outbreak.

Crystal structure of human prion protein bound to a therapeutic antibody.

Prion infection is characterized by the conversion of host cellular prion protein (PrP(C)) into disease-related conformers (PrP(Sc)) and can be arrested in vivo by passive immunization with anti-PrP monoclonal antibodies. Here, we show that the ability of an antibody to cure prion-infected cells correlates with its binding affinity for PrP(C) rather than PrP(Sc). We have visualized this interaction at the molecular level by determining the crystal structure of human PrP bound to the Fab fragment of monoclonal antibody ICSM 18, which has the highest affinity for PrP(C) and the highest therapeutic potency in vitro and in vivo. In this crystal structure, human PrP is observed in its native PrP(C) conformation. Interactions between neighboring PrP molecules in the crystal structure are mediated by close homotypic contacts between residues at position 129 that lead to the formation of a 4-strand intermolecular beta-sheet. The importance of this residue in mediating protein-protein contact could explain the genetic susceptibility and prion strain selection determined by polymorphic residue 129 in human prion disease, one of the strongest common susceptibility polymorphisms known in any human disease.

Modified Heider balance on Erdös-Rényi networks.

The lack of signed random networks in standard balance studies has prompted us to extend the Hamiltonian of the standard balance model. Random networks with tunable parameters are suitable for better understanding the behavior of standard balance as an underlying dynamics. Moreover, the standard balance model in its original form does not allow preserving tensed triads in the network. Therefore, the thermal behavior of the balance model has been investigated on a fully connected signed network recently. It has been shown that the model undergoes an abrupt phase transition with temperature. Considering these two issues, we examine the thermal behavior of the structural balance model defined on Erdös-Rényi random networks within the range of their connected regime. We provide a mean-field solution for the model. We observe a first-order phase transition with temperature for a wide range of connection probabilities. We detect two transition temperatures, T_{cold} and T_{hot}, characterizing a hysteresis loop. We find that with decreasing the connection probability, both T_{cold} and T_{hot} decrease. However, the slope of decreasing T_{hot} with decreasing connection probability is larger than the slope of decreasing T_{cold}. Hence, the hysteresis region gets narrower until it disappears in a certain connection probability. We provide a phase diagram in the temperature-tie density plane to accurately observe the metastable or coexistence region behavior. Then we justify our mean-field results with a series of Monte Carlo simulations.

Effect of pre-treatment of ovine sperm on male pronuclear formation and subsequent embryo development following intracytoplasmic sperm injection.

The objective of this study was to determine the effects of various methods of sperm pre-treatment on male pronuclear (MPN) formation and subsequent development of ovine embryos derived from in vitro-matured oocytes and intracytoplasmic sperm injection (ICSI). The effect of treatment of injected oocytes with dithiothreitol (DTT) on embryo development was also assessed. In Exp. 1, the injected oocytes with non-treated sperm were activated with three different procedures. The cleavage and blastocyst rates in those activated with DTT was lower (p<0.05) than those activated with either ionomycin (Io) +6-dimethylaminopurine (6-DMAP) or DTT + I + 6-DMAP. In Exp. 2, the effects of sperm pre-incubated with DTT, sodium dodecyl sulphate (SDS) or DTT + SDS as well as two-time frozen/thawed sperm (without cryoprotectant) on MPN formation and oocyte activation were examined. The non-treated sperm served as controls. The MPN formation in DTT + SDS group was higher (p<0.05) than other groups except for freeze-thaw group. No difference in the rate of activated ICSI oocytes was observed among groups. In Exp. 3, the effect of pre-treatment of sperm on subsequent development of ICSI embryos and blastocyst cell numbers were examined. The rates of cleavage and blastocyst formation as well as the blastocyst cell numbers were similar among the pre-treated and control groups. In conclusion, pre-treatment of sperm with DTT + SDS positively affected MPN formation, although the subsequent development capacity of the resulting embryos remained limited. Moreover, DTT was not effective on oocyte activation compared with Io + 6-DMAP after ICSI.

Heider balance under disordered triadic interactions.

The Heider balance addresses three-body interactions with the assumption that triads are equally important in the dynamics of the network. In many networks, the relations do not have the same strength, so triads are differently weighted. Now, the question is how social networks evolve to reduce the number of unbalanced triangles when they are weighted? Are the results foreseeable based on what we have already learned from the unweighted balance? To find the solution, we consider a fully connected network in which triads are assigned with different random weights. Weights are coming from Gaussian probability distribution with mean µ and variance σ. We study this system in two regimes: (I) the ratio of µ/σ≥1 corresponds to weak disorder (small variance) that triads' weight are approximately the same; (II) µ/σ<1 counts for strong disorder (big variance) and weights are remarkably diverse. Investigating the structural evolution of such a network is our intention. We see disorder plays a key role in determining the critical temperature of the system. Using the mean-field method to present an analytic solution for the system represents that the system undergoes a first-order phase transition. For weak disorder, our simulation results display the system reaches the global minimum as temperature decreases, whereas for the second regime, due to the diversity of weights, the system does not manage to reach the global minimum.

Vitrification of in vitro produced ovine embryos at various developmental stages using two methods.

This study was conducted to evaluate the effects of developmental stage of in vitro produced (IVP) ovine embryos and the type of vitrification procedure used on embryo cryotolerance. The IVP embryos were vitrified at five different developmental stages: 4-, 8- and 16-cell, morula, and blastocyst. For each stage, half of the embryos were vitrified in either 30 microl 3.4M glycerol+4.6M ethylene glycol in straw (method 1) or in <0.1 microl 2.7 M ethylene glycol+2.1 M Me(2)SO+0.5M sucrose placed on the inner surface of a straw (method 2) of vitrification solution, based on two different procedures. After warming embryo viability was determined by assessing the rates of re-expansion, survival, and blastocyst formation. The quality of surviving embryos was evaluated by their hatching rate and blastocyst cell numbers. In both vitrification methods, embryo survival progressively increased as the developmental stage progressed. In method 1 few of the early cleavage stage embryos (4-, 8- and 16-cell) could reach to the blastocyst stage following warming. There was no significant difference in blastocyst cell numbers (total, ICM, and trophectoderm cells) or hatching rate of blastocysts derived from vitrified embryos at different developmental stages. The number of dead cells in vitrified blastocysts in method 1 was higher than for non-vitrified blastocysts (P<0.05). The number of apoptotic cells in vitrified blastocysts was higher than for non-vitrified counterparts (P<0.05). In conclusion, both the developmental stage of IVP ovine embryos and the method of vitrification have a significant effect on the viability and developmental competence of sheep embryos.

Effect of donor cell age on development of ovine nuclear transfer embryos in vitro.

The effects of the age of cell donor animal on in vitro development of ovine nuclear transfer (NT) embryos were investigated. Somatic donor cells were obtained from two different sources: (1) adult cells (adult fibroblast cells; AFC and adult cumulus cells; ACC); and (2) fetal fibroblasts (40-day-old; FFC-40 and 65-day-old; FFC-65). The fibroblast cell lines were used for NT procedures within 4-13 subpassages. While the cumulus cells were used as non-cultured (fresh) cells. The in vitro matured abattoir-derived oocytes were considered as recipients. No differences in the rates of fusion (75.7, 77.7, 76.3 and 86.7%) and cleavage (80.1, 84.3, 77.8 and 74%) were detected among couplets reconstructed with FFC-40, FFC-65, AFC and ACC, respectively. Blastocyst formation rate of those oocytes reconstructed with FFC-40 was higher (18%; p < 0.001) than those reconstructed with FFC-65 (13%) and AFC (10.9) and comparable with those reconstructed with ACC (17.5%). When the effect of passage number was analysed within groups (FFC-40, FFC-65 and AFC) there were no significant differences in fusion, cleavage and blastocyst rates between reconstructed oocytes. The present study demonstrates that the fetal and adult fibroblasts as well as fresh cumulus cells are comparable in their ability to attain cell fusion and embryonic cleavage. Moreover, the blastocyst formation rate is influenced by the age of the donor animal and the fresh cumulus cells have similar remodelling potential to that of fetal fibroblasts in term of blastocyst formation rate.

Effects of growth hormone on nuclear maturation of ovine oocytes and subsequent embryo development.

The objective of this study was to determine the effect of the presence of recombinant ovine growth hormone either alone or together with follicle stimulating hormone (FSH) during ovine oocyte in vitro maturation (IVM) on nuclear maturation and subsequent embryo development. Moreover, the effect of growth hormine (GH) on embryo development whether influenced by the presence of foetal bovine serum (FBS) was assessed. The abattoir-derived oocytes were randomly divided into four treatment groups and cultured in maturation medium supplemented with: (i) 0.05 IU/ml FSH; (ii) 300 ng/ml roGH; (iii) FSH + roGH; and (iv) no FSH and GH (control). The percentages of germinal vesicle-stage oocytes in GH-treated group after 8 h of culture was significantly higher than the FSH and FSH + GH groups and lower than control (22.4%, 8.7%, 9.1%, and 32% respectively). The percentage of MII-stage oocytes was significantly increased in the presence of GH after 16 and 24 h of culture compared to the control (44.7% and 83.1% vs 32.6% and 73.6% respectively). There was no significant synergism between GH and FSH in terms of nuclear maturation. The blastocyst rates in serum-supplemented groups were enhanced by the presence of FSH and GH compared to the control (35.4% and 31.3 vs 11.4% respectively). Compared with either GH or FSH alone, the subsequent embryo development (blastocyst rate), however, was negatively influenced by co-presence of both hormones (22.8%). In contrast, the corresponding values were not affected in the absence of serum. In conclusion, GH had positive effect on nuclear maturation of sheep oocytes. Moreover, the pattern of the effect of GH on embryo development was influenced by the presence of FBS during IVM.

Morphological and molecular identification of Corynosoma caspicum, and its histopathological effect on the intestinal tissue of a Caspian seal (Pusa caspica).

BACKGROUND: Corynosoma is a parasite from the Acanthocephala phylum with worldwide distribution. Corynosoma parasites are found in pinnipeds as their definitive host. AIMS: This study aimed to investigate the morphological and molecular characteristics of Corynosoma, and its histopathological effect on the intestinal tissue of Pusa caspica. METHODS: A severe Corynosoma infection was observed in the small intestine of a juvenile male Caspian seal (P. caspica). The morphological descriptions were done using light microscopy, and scanning electron microscopy (SEM). The molecular diagnosis was performed using partial sequences of internal transcribed spacer 1 (ITS1) and 5.8S ribosomal RNA (rRNA) gene. RESULTS: According to the results, the Corynosoma specimens were identified as Corynosoma caspicum. The histopathological inspection of intestinal tissue revealed lesions in epithelial cells, mucosa, submucosa and muscle layers, destruction of intestinal glands, and infiltration of inflammatory cells. CONCLUSION: Presence of such a severe infection in one of the individual Caspian seals can suggest the possibility of morbidity among other seals in the landlocked Caspian Sea. Thus, further research on their parasite infections is required for understating the status of the Caspian seal population and conserving this endangered species.

Effect of culture system on survival rate of vitrified bovine embryos produced in vitro.

This study was designed to evaluate the effect of in vitro culture system on bovine blastocyst yield and quality after vitrification. In Experiment 1, IVM/IVF zygotes were allocated to three culture conditions: (I) Oviductal cells-SOF (OCM-SOF); (II) Oviductal cells-TCM (OCM-TCM); and (III) SOF for 8 days. There was no significant difference between blastocyst rates among groups. In Experiment 2, the IVP-blastocysts in three above culture conditions were vitrified within groups segregated according to age (Day 7 and 8) and blastocoelic cavity size (early and expanded blastocysts). A trend of higher survival rate was obtained in vitrified/warmed early blastocysts compared with expanded ones, so that the difference in OCM-TCM group was significant (P<0.001). Higher survival and hatching rates (P<0.001) were obtained in OCM-SOF and OCM-TCM groups (co-culture) compared with SOF group and the age of blastocyst had no effect on post-thaw survival and hatching rates. In Experiment 3, after staining of blastocysts, in fresh blastocysts the highest number of trophectoderm cells was observed in OCM-TCM group and the number of inner cell mass (ICM) was higher in co-culture groups than SOF group (P<0.001). In vitrified/warmed blastocysts the number of ICM and trophectoderm cells in co-culture groups was higher than SOF group (P<0.001) except for the ICM of expanded blastocysts. In conclusion, in our culture conditions, the blastocyst yield is not influenced by culture system, while the cryotolerance of IVP-blastocysts is positively influenced by the presence of somatic cells. Moreover, the expanded blastocysts are more susceptible to cryoinjury than early blastocysts.

In vitro developmental competence of ICSI-derived activated ovine embryos.

The present study was conducted to determine the necessity for activation after intracytoplasmic sperm injection (ICSI) in sheep. The effect of chemical stimulation with either 5microM ionomycin (I) for 5min or ionomycin+2mM 6-dimethylaminopurine (6-DMAP) for 3h on the efficiency of ICSI, was compared in six experimental groups: (1) ICSI, (2) ICSI+I, (3) ICSI+I+6DMAP, (4) Sham, (5) Sham+I, and (6) parthenogenetics (Sham and parthenogenetic groups were used as controls). In the present study, ovine oocytes needed additional chemical stimulation, after conventional ICSI, to activate (female pronucleous formation) and to form zygotes with male and female pronuclei (2PN). The percentage of cleaved embryos obtained and developed to blastocyst stage was higher (P<0.001) for ICSI-derived zygotes, followed by activation (I and I+6DMAP; 18.2 and 22.5%, respectively) than ICSI and Sham injection without activation (3.0 and 0.0%, respectively). There was, however, no significant difference between activation protocols I or I+6DMAP. Furthermore, there was no significant difference among chemically activated, ICSI-derived zygotes in term of hatchability rate; however, the percentage was significantly higher in parthenogenetic and IVF groups than ICSI and Sham injection. In conclusion, neither sperm alone nor mechanical activation was sufficient for ovine oocyte activation and pronuclei formation. Therefore, in our study conditions for in vitro embryo development, chemical activation of oocytes must be considered an essential part of the ICSI procedure in sheep.

Sirtuin inhibition and anti-cancer activities of ethyl 2-benzimidazole-5-carboxylate derivatives.

New benzimidazoles were synthesized based on the previously identified sirtuin inhibitor BZD9L1. The compounds were screened for their sirtuin (SIRT1, SIRT2 and SIRT3) inhibitory activities. Compound BZD9Q1 was determined to be a pan-SIRT1-3 inhibitor. Furthermore, the proliferation of various cancer cells was inhibited by BZD9Q1. It was shown that BZD9Q1 elicits a cytostatic effect by inducing cell cycle arrest at the G2/M phase while also showing a prominent induction of apoptosis against oral cancer cells.