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Various prenatal factors including the number of littermates (fraternity size) and exposure to male littermate (fraternity sex ratio) during fetal period have been reported to influence postnatal fertility in the mammals. The present research was conducted to study the association of fraternity size and sex ratio with reproductive performance of nulliparous ewes and does. To this end, data associated with number of littermates, exposure to male littermate, birth weight, age at first pregnancy, as well as litter size, sex ratio of offspring, litter weight, and birth weight of female and male offspring after the first parturition retrieved from the database of sheep (n = 536 Romane and 289 Blanche du Massif Central ewes) and goat (n = 174 Alpine and 267 Saanen does) flocks. Fraternity size was negatively associated with birth weight of ewes and does (P < 0.05). Exposure to male littermate during fetal period was associated with younger age at first pregnancy and larger litter size in the does (P < 0.05), but not in the ewes (P > 0.05). Exposure to male littermate during fetal period was positively associated with the odds of male-biased litters in the ewes and does (P < 0.05). Fraternity size was positively associated with litter weight in the does (P < 0.05), but not in the ewes (P > 0.05). In conclusion, the present study showed that the number and sex of littermates during fetal period could impact postnatal reproduction of ewes and does. In this context, some associations, particularly those related to exposure to male littermate during fetal period, were only observed in does, which implicates that the effect of androgens on developmental programming of reproduction may be species-specific., Competing Interests: Declaration of Competing Interest The authors have no conflict of interest to disclose., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Level of dam milk production (DMP) and dam milk fat to protein ratio (DFPR), as an indicator of metabolic status in dairy cows, have been identified to be associated with productive and reproductive performance of the offspring. Yet whether the interaction of DMP by DFPR can be associated with performance of the offspring have not been studied to our knowledge. Therefore, the present study was conducted to investigate the association of the main and interactive effects of DMP and DFPR with offspring's birth weight, survival, milk yield and fertility. To this end, data of birth weight, culling rate, milk yield and reproductive variables of offspring born to lactating dams (n = 14,582) and data associated with DMP and DFPR during 305-day lactation were retrieved. Afterwards, offspring were classified in three categories of DMP, including DMP1 (dams with <10.00 × 10 3  kg of 305-day milk production), DMP2 (dams with ≥10.00 × 10 3  kg and <14.00 × 10 3  kg of 305-day milk production), DMP3 (dams with ≥14.00 × 10 3  kg of 305-day milk production), and three categories of DFPR, including DFPR1 (offspring born to dams with <1.00 FPR), DFPR2 (offspring born to dams with ≥1.00 and < 1.40 FPR) and DFPR3 (offspring born to dams with ≥1.40 FPR). Statistical analysis revealed no association of the interaction effect of DMP by DFPR with investigated variables in the offspring (P > 0.05). However, the main effect of DMP was positively associated with milk yield, but negatively associated with survival, age at first insemination and conception during nulliparity, and transgenerational improvement in milk yield in the offspring (P < 0.05). Moreover, the main effect of DFPR was positively associated with birth weight, survival and first service conception rate during nulliparity, but negatively associated with metabolic status and reproductive performance during primiparity in the offspring (P < 0.05). In conclusion, the present study did not find any interaction effect of DMP by DFPR on productive and reproductive variables in the offspring. This finding implicates the association of DMP with milk production in the offspring was regardless of DFPR. Moreover, this finding implies the association of DFPR with postpartum metabolic status and reproductive performance in the offspring was regardless of DMP., Competing Interests: Declaration of competing interest The authors have no conflict of interest to disclose., (Copyright © 2024 Elsevier Inc. All rights reserved.)

Although the association of maternal milk production with developmental programming of offspring has been investigated, there is limited information available on the relationship of maternal milk components with productive and reproductive performance of the offspring. Therefore, the present study was conducted to analyze the association of maternal milk fat and protein percentage and milk fat to protein ratio with birth weight, survival, productive and reproductive performance and AMH concentration in the offspring. In study I, data of birth weight, milk yield and reproductive variables of offspring born to lactating dams (n = 14,582) and data associated with average maternal milk fat percentage (MFP), protein percentage (MPP) and fat to protein ratio (MFPR) during 305-day lactation were retrieved. Afterwards, offspring were classified in various categories of MFP, MPP and MFPR. In study II, blood samples (n = 339) were collected from offspring in various categories of MFP, MPP and MFPR for measurement of serum AMH. Maternal milk fat percentage was positively associated with birth weight and average percentage of milk fat (APMF) and protein (APMP) and milk fat to protein ratio (FPR) during the first lactation, but negatively associated with culling rate during nulliparity in the offspring (P < 0.05). Maternal milk protein percentage was positively associated with birth weight, APMF, APMP, FPR and culling rate, but negatively associated with milk yield and fertility in the offspring (P < 0.05). Maternal FPR was positively associated with APMF and FPR, but negatively associated with culling rate, APMP and fertility in the offspring (P < 0.05). However, concentration of AMH in the offspring was not associated with MFP, MPP and MFPR (P > 0.05). In conclusion, the present study revealed that maternal milk fat and protein percentage and their ratio were associated with birth weight, survival, production and reproduction of the offspring. Yet it was a preliminary research and further studies are required to elucidate the mechanisms underlying these associations., Competing Interests: Declaration of competing interest The authors have no conflict of interest to disclose., (Copyright © 2024 Elsevier Inc. All rights reserved.)

Although some studies investigated the relationship of dam milk production (DMP) with offspring birth weight and productive performance, limited information is available on the association of level of DMP with reproductive performance in dairy cows. Therefore, the present study was conducted to understand whether dams with various levels of milk production produce offspring with different fertility. In study I, offspring were classified based on the level of DMP into five categories, including DMP1 (dams with <10.00 × 10 3  kg of 305-day milk production), DMP2 (dams with ≥10.00 × 10 3  kg and <12.00 × 10 3  kg of 305-day milk production), DMP3 (dams with ≥12.00 × 10 3  kg and <14.00 × 10 3  kg of 305-day milk production), DMP4 (dams with ≥14.00 × 10 3  kg and <16.00 × 10 3  kg of 305-day milk production) and DMP5 (dams with ≥16.00 × 10 3  kg of 305-day milk production). In study I, data of birth weight, milk yield and reproductive variables of 14,536 offspring born to lactating dams and corresponding data of DMP were retrieved. In study II, blood samples (n = 339) were collected from offspring in various categories of DMP for measurement of serum AMH. Offspring were heavier at birth in DMP4 and DMP5 categories than DMP1 and DMP2 categories (P < 0.05). Milk yield of offspring increased as DMP elevated (P < 0.05); however, offspring in DMP1 and DMP2 categories produced higher milk as compared with their dams during primiparity (P < 0.05) whereas offspring in DMP3, DMP4 and DMP5 categories produced less milk as compared with their dams during primiparity (P < 0.05). Milk fat to protein ratio during the first month of lactation was greater in DMP4 and DMP5 categories than DMP1 category (P < 0.05). Offspring of DMP4 and DMP5 categories were inseminated and conceived at younger ages than offspring of DMP1 category during nulliparity (P < 0.05). Calving to conception interval was longer in DMP5 than DMP1 category in primiparous offspring (P < 0.05), but concentration of AMH did not differ among various categories of DMP (P > 0.05). In conclusion, dams with greater level of milk production produced heavier offspring with higher milk yield but worse transgenerational improvement in milk production and diminished reproductive performance, which were seemingly under higher pressure of negative energy balance during the first month of lactation., Competing Interests: Declaration of competing interest The authors have no conflict of interest to disclose., (Copyright © 2024 Elsevier Inc. All rights reserved.)

Although the negative effect of maternal exposure to heat stress on production and reproduction of offspring has been reported, there are some discrepancies among various studies about which gestational stage is more critical in this regard. Therefore, the present research was conducted to identify during which stage(s) of pregnancy maternal exposure to heat stress would lead to more dramatic decrease in productive and reproductive performance of offspring. To this end, offspring were classified based on the gestational stage they were in utero exposed to heat stress into four categories, including heat stress exposure (HSE) during only the first trimester of gestation (HSE1), HSE during the first and second trimester of gestation (HSE2), HSE during the second and third trimester of gestation (HSE3) and HSE during only the third trimester of gestation (HSE4). In study I, data of birth weight, milk yield and reproductive variables of 11,788 offspring and data of the month they were conceived were retrieved. In study II, blood samples (n = 521) were collected from offspring in various categories of HSE for measurement of serum AMH. Offspring in HSE1 and HSE2 categories were heavier than offspring in HSE3 and HSE4 categories (P < 0.0001). Offspring in HSE1 and HSE3 categories had the lowest and highest milk production, respectively (P < 0.05). First service conception rate was the greatest and worst in HSE1 and HSE4 categories, respectively (P < 0.05). Service per conception and calving to conception interval were greater in HSE2 than HSE4 category (P < 0.05). Concentration of AMH was lower in HSE1 than HSE4 category (P < 0.05). In conclusion, the present study indicated that the early stage of gestation could be a more critical period for the negative impact of in utero heat stress on developmental programming of milk production and ovarian reserves. Yet an evident temporal pattern for the adverse effect of maternal heat stress on developmental programming of reproductive performance in offspring was not found., Competing Interests: Declaration of competing interest The authors have no conflict of interest to disclose., (Copyright © 2023 Elsevier Inc. All rights reserved.)

Multiparous dams have been reported to produce offspring with greater fertility and higher AMH concentration, as a marker of ovarian reserves, as compared with nulliparous and primiparous dams. Yet it has remained to be addressed whether this phenomenon can still be true for old multiparous cows which might experience some geriatric changes in their reproductive system. Therefore, the present study was conducted to evaluate the productive and reproductive performance of offspring with different maternal parity. To this end, offspring were classified based on their maternal parities into four categories, including offspring of nulliparous (no previous parity), primiparous (one previous parity), young multiparous (two to six previous parities) and old multiparous (seven or more previous parities) dams. In study I, data of birth weight, milk yield and reproductive variables of 11,788 offspring and data of their maternal parity were retrieved. In study II, blood samples (n = 521) were collected from offspring with various maternal parity for measurement of serum AMH. Birth weight was the lowest in the offspring of nulliparous dams (P < 0.0001) and it was lower in offspring of primiparous and old multiparous dams than offspring of young multiparous dams (P < 0.05). Milk production was the lowest in offspring of old multiparous dams (P < 0.01), and it was lower in offspring of young multiparous dams than offspring of nulliparous and primiparous dams (P < 0.0001). Offspring of old multiparous dams had greater first service conception rate, less services per conception and shorter calving to conception interval than offspring of nulliparous, primiparous and young multiparous dams (P < 0.05). Furthermore, AMH concentration was higher in offspring of old multiparous dams than offspring of nulliparous and primiparous dams (P < 0.05). In conclusion, the present study revealed greater milk production in offspring resulting from dams with lower parity, probably due to the genetic selection for improvement of milk production in dairy cows which imparts the younger generations greater genetic merits for milk production. Reproductive performance, however, was greater in offspring born to dams with higher parity, particularly those born to old multiparous dams, and this phenomenon might be related to their lower milk production and higher AMH concentration., Competing Interests: Declaration of competing interest The authors have no conflict of interest to disclose., (Copyright © 2023 Elsevier Inc. All rights reserved.)

Unlike GnRH, estradiol could induce emergence of a new follicular wave regardless of the size of follicle. Therefore, the present study was conducted to understand whether replacement of the first GnRH by estradiol in the breeding protocol of Double Ovsynch program could enhance fertility. Cows were randomly assigned to two groups, including Double Ovsynch protocol (Control; n = 120) and Ovsynch-estradiol-PGF2α-GnRH (EPG) protocol (Treatment; n = 120). Cows in both groups were subjected to presynchronization Ovsynch. Seven days later, cows in the control group received GnRH, which was followed by PGF2α and GnRH 7 days and 9 days plus 8 h later, respectively. Cows in treatment group received estradiol 7 days after the second GnRH of presynchronization Ovsynch, which was followed by PGF2α and GnRH 7 days and 10 days plus 8 h later, respectively. Cows were subjected to timed AI (TAI) 16 h after final GnRH in both groups. Pregnancy per AI (P/AI) was greater in cows in treatment than control group (64.17 % vs. 44.17 %, respectively; P = 0.02). Cows with a follicle with diameter ≥ 10 mm (F10) at the beginning of EPG in treatment group had greater P/AI than cows without a F10 at the beginning of breeding Ovsynch in control group (P ≤ 0.05). Pregnancy per AI was greater in cows with a CL at the beginning of EPG in treatment group than cows without a CL at the same timepoint in treatment group, and cows with or without a CL at the beginning of breeding Ovsynch in control group (P ≤ 0.05). In conclusion, inclusion of estradiol in Double Ovsynch protocol as a replacement for the first GnRH of breeding Ovsynch could improve fertility, particularly in cows with a CL at the initiation of EPG., Competing Interests: Conflict of interest statement The authors have no conflict of interest to disclose., (Copyright © 2023 Elsevier B.V. All rights reserved.)

Although the effect of mastitis on reproduction and production of lactating dairy cows has been vastly studied, little information is available about the association of maternal udder health status with offspring reproduction and production. Therefore, the present research was conducted to study the association between maternal average monthly somatic cell count (SCC) with reproduction, anti-Müllerian hormone (AMH) concentration, udder health status and milk production in the offspring. Based on maternal average monthly SCC (MSCC), offspring were classified into five categories including MSCC1 (SCC <200,000; n = 3005), MSCC2 (200,000 ≤ SCC <400,000; n = 252), MSCC3 (400,000 ≤ SCC <600,000; n = 103), MSCC4 (600,000 ≤ SCC <800,000; n = 40) and MSCC5 (SCC ≥800,000; n = 61). Data associated with reproduction, production and udder health status of offspring were retrieved from the herd database. In addition, blood samples were collected from a subset of offspring (n = 136) for measurement of serum AMH, as a reliable marker of ovarian reserves. Offspring in MSCC5 category had more services per conception and longer calving to conception interval than offspring in MSCC1 and MSCC2 categories (P < 0.05). The average number of SCC and risk of clinical mastitis in the offspring were not associated with MSCC (P > 0.05). But offspring in MSCC5 category produced less milk, fat and protein than offspring in MSCC1 category (P < 0.05). In addition, AMH concentration was lower in MSCC5 than MSCC1 category (P < 0.05). In conclusion, the present study showed that elevated maternal average monthly SCC could culminate in birth of offspring with inferior reproductive performance, smaller size of ovarian reserves and lower level of milk production during the first lactation period., Competing Interests: Declaration of competing interest The authors have no conflict of interest to disclose., (Copyright © 2023 Elsevier Inc. All rights reserved.)

Serial progesterone injections followed by human menopausal gonadotropin (hMG), instead of equine chorionic gonadotropin (eCG), were used to synchronize estrus in ewes. Shal ewes (n = 189) were assigned into five groups and each group was divided into two sub-groups to receive gonadotropins including eCG (300 IU; intra-muscular) or hMG (one ampoule; subcutaneously, SC). All ewes received prostaglandin (PG) F 2 α six days after introducing ram (day 0). Ewes received 0 (control), one, two, three or four injections of progesterone (50.00 mg; SC), 72 hr apart. The first progesterone was injected at the time of PG injection. Ewes in treatment groups received gonadotropins 48 hr after the last progesterone injection. Control group ewes received gonadotropins, at the time of PG injection. Mating was recorded after introducing fertile rams. Data were analyzed using GLM and GENMOD procedures in SAS. The incidence of estrus was less in control and ewes received a single progesterone (34.20%) compared to ewes received two (64.10%), three (81.10%) and four injections (68.40%) of progesterone. Time to estrus was earlier in control (45.70 ± 4.41 hr) than progesterone-treated groups (63.60 ± 1.79 hr). Fertility (51.30%) and fecundity (78.40%) of ewes received three progesterone injections were significantly greater than other progesterone-treated groups. There was no significant difference in reproductive indices between eCG and hMG sub-groups. In conclusion, during the non-breeding season, three injections of progesterone, three days apart, starting six days after ram exposure, in association with hMG, 48 hr after the last progesterone injection, could provide a sound reproductive performance in Shal ewes., Competing Interests: The authors have no conflicts of scientific interest with respect to the manuscript., (© 2023 Urmia University. All rights reserved.)

Anogenital distance (AGD) is related to fertility in cows, but there is little information on the association of AGD and reproduction in does. Therefore, the present study was conducted to assess the relationship between AGD and reproductive variables in Murciano-Granadina does. AGD was measured as the distance between anus and clitoris and does (n = 578) were divided into two categories of AGD, including short AGD (AGD ≤ median of AGD in studied does; n = 313) and long AGD (AGD > median of AGD in studied does; n = 265). Data of reproductive variables were retrieved from the herd database and were analyzed using multivariable statistical models, in which the potential confounding factors were included. AGD data were not normally distributed (3.78 ± 0.02 cm) and ranged from 2.70 to 5.10 cm. AGD was longer in multiparous and primiparous does than nulliparous does (P < 0.0001), and was positively associated with age and body weight (P < 0.0001). The risk of pregnancy and kidding, litter size, fecundity, proportion of male offspring and birth weight of kids were higher in long AGD does than short AGD does (P < 0.05). But abortion risk did not differ between does with long and short AGD (P > 0.05). In conclusion, the present study revealed that AGD had individual variation among Murciano-Granadina does, and does with long AGD were more fertile and more likely to produce larger, heavier and male-biased litters as compared with does with short AGD., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Toxocara canis (T. canis) is a common parasitic nematode in dogs and cats. Parasitic worms can cause chronic and long-term infections in their host, due to their ability to neutralize the host's defense mechanisms. They can stimulate immune response-mediated regulatory T (T-reg) cells. The aim of this study is to evaluate the effect of recombinant T. canis C-type lectin protein (TCTL-1) on cell infiltration in the brains of BALB /c mice as well as the number of regulatory T cells. Six-week-old female BALB/c mice received the recombinant C-type lectin protein of T. canis six times intravenously and intraperitoneally. Twenty-eight days after the first injection, the spleen and brain of mice were removed under sterile conditions. The brains of mice were examined by histopathological staining methods. The FOXP3+ regulatory T cell population was determined by flow cytometry. The cell populations of regulatory T cells in spleen mononuclear cell culture of 3 female BALB/c mice injected with recombinant TCTL-1 (group I) were 2.59%, 1.64%, and 1.78 and in spleen mononuclear cell culture of three female BALB/c mice injected with sterile PBS (group II) as a control group were 1.14%, 1.13%, and 1.15%. Also, no cell infiltration was seen around the cerebral arteries of mice receiving this protein. This recombinant protein would increase the population of FOXP3+ regulatory T cells. These results suggest that recombinant C-type lectin protein of T. canis can modulate immune responses, reduce severe inflammatory responses, and induce FOXP3+ regulatory T cells.

There was investigation of whether there were ovulations from co-dominant follicles following eCG administration. In all experiments, there was GnRH injection and CIDR insertion on day 0 (D0), CIDR withdrawal on D8, and cloprostenol administration on D8 (Exp. I and II) or D7 and D8 (Exp. III). Females in the control group were not administered any further treatment. Females in other group(s) were treated with eCG (500 IU) on Day 2 in Exp. I, Day 2 (eCG-2) or 8 (eCG-8) in Exp. II and Day 2 (eCG-2) or Days 2 and 6 (eCG-2-6) in Exp. III. Ovaries were examined using ultrasonography. In Experiments I and II, females had follicle emergence on Day 2. At the time of CIDR removal, more eCG-treated heifers (8/9; Exp. I) and cows (5/6; eCG-2; Exp. II) had co-dominant follicles compared to those in the control group (P < 0.05). Occurrence of ovulations from co-dominant for individual cows was minimal. In Experiment III, the time period from CIDR removal to estrus in cows treated with eCG-2 (68 ± 13 h) was longer compared to cows in the control (37±2 h) and eCG-2-6-treated group (38 ± 5 h; P < 0.05). There was a greater proportion of heifers having ovulations and thus greater progesterone concentration in the eCG-2-6 than eCG-2 group (P < 0.05). Administering eCG twice 4 days apart with the initial administration being two days after GnRH administration, at the time of follicle wave emergence, could induce growth of and ovulation from co-dominant follicles and enhance progesterone production in cattle., (Copyright © 2020 Elsevier B.V. All rights reserved.)

In cattle, results of some but not other studies indicated there has been an association of circulating anti-Müllerian hormone (AMH) concentration with reproductive variables. Considering the inconsistency in results among previous studies, there was evaluation of associations between circulating AMH and reproductive performance in dairy cows, and to this end, the prevailing data of serum AMH concentration and reproductive variables were used (n = 172). Preliminarily, association of AMH concentration with reproductive indices were analyzed using linear and quadratic models. Subsequently, cows were categorized based on AMH concentrations in four equal quartiles, including cows with the least (Q 1 ; n = 43), moderately lesser (Q 2 ; n = 43), moderately greater (Q 3 ; n = 43) and greatest (Q 4 ; n = 43) AMH concentrations, and reproductive variables were compared among the respective AMH quartiles. Initially, assessments indicated reproductive variables were not linearly associated with serum AMH (P > 0.05); however, for all reproductive variables except days to first breeding service (DFS) there was a quadratic association with serum AMH (P < 0.05). Results from analysis of reproductive performance in various AMH quartiles indicated there was a longer period of DFS for cows in Q 3 than Q 1 (P < 0.05). First postpartum insemination to conception and calving to conception intervals were shorter for cows in Q 3 than Q 1 and Q 4 (P < 0.05) and for cows in Q 2 than Q 1 (P < 0.05). In conclusion, results from the present study indicate cows with intermediate AMH concentrations had greater reproductive performance., Competing Interests: Declaration of Competing Interest The authors affirm that there is no conflict of interest., (Copyright © 2020 Elsevier B.V. All rights reserved.)

The objective of this study was to investigate the effect of equine chorionic gonadotropin (eCG) on ovarian follicles at three stages of development (emergence, dominance and early static phases) during the first follicular wave (FFW) in Holstein heifers. Heifers (n=20) were randomly assigned into four experimental groups (n=5 in each group). Heifers received eCG (500 IU; Folligon ® ; Intervet, Holland; i.m) a) on the day of follicle emergence (day of ovulation; group 1), b) on the dominant phase (dominant follicle (DF): the first day in which follicle was observed at ≥10 mm; group 2, and c) on the early static phase (group 3) of the FFW. Control group heifers did not receive any treatment. Daily ultrasonography was conducted to monitor ovarian structure throughout estrous cycle. All treatment group heifers, regardless of the stage of follicle development, displayed follicle growth after eCG injection. Administration of eCG, in group 1, hastened DF detection and induced co-dominant follicles; whereas, in groups 2 and 3, it delayed DF regression, and increased cycle length compared to control. In all treatment group heifers, DF was present 84 h after eCG injection. Maximum diameter of corpus luteum was larger in eCG treated groups compared to control (P<0.05). In conclusion, depending on the time of eCG administration throughout the FFW (emergence, dominant and early static phases), co-dominancy, maintenance of DF, enhancement of follicle and corpus luteum growth and increase in estrous cycle length could be observed in Holstein heifers.

Nutritional partitioning to fetal development differ among nulliparous, primiparous and multiparous cows, leading to birth of smaller calves to nulliparous and primiparous than multiparous dams. The prenatal nutritional state could influence ovarian reserve and fertility in the adult offspring. The effect of maternal parity on ovarian reserve and reproduction of female offspring, however, has not been investigated in cattle. Accordingly, a retrospective research was conducted to study reproductive variables in nulliparous- (n = 310), primiparous- (n = 236) and multiparous-born (n = 323) female offspring during the first four lactation periods in dairy cows. Additionally, anti-Müllerian hormone (AMH), as a reliable marker of ovarian reserve, was measured in a subset of nulliparous- (n = 51), primiparous- (n = 51) and multiparous-born (n = 49) female offspring. Birth weight in multiparous-born calves was greater than that in nulliparous- and primiparous-born calves (P <  0.01). Days to first service was shorter in multiparous-born female offspring as compared with nulliparous- and primiparous-born female offspring (P <  0.05). First service conception rate, proportion of repeated breeders, service per conception and calving to conception interval were greater in multiparous-born than nulliparous-born cows (P <  0.05). Cumulative culling rate was greater in primiparous-born cows than nulliparous- and multiparous-born cows (P <  0.05). The AMH concentration was greater in multiparous-born than nulliparous- and primiparous-born cows (P <  0.05). In conclusion, the present study revealed that in comparison with multiparous cows, nulliparous and primiparous cows, particularly nulliparous cows, produce female offspring with lesser reproductive performance and smaller size of ovarian reserves., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Regulatory T cells (Tregs) play a major role in the prevention of autoimmune diseases. Transfer of Foxp3 gene into conventional T cells converts their phenotype to regulatory T cells. Therefore, the question arises as to whether adoptively transferred in vitro differentiated Treg cells specific for a locally expressed antigen might have better inhibitory effects on the progression of the disease as compared with antigen-nonspecific T reg cells. Herein, we investigated the therapeutic potential of primed and unprimed retrovirus mediated Foxp3-overexpression T cells following intravenously injected of these cells into affected rats with collagen-induced arthritis (CIA), an animal model of rheumatoid arthritis. Our analyses demonstrate that systemic administration of collagen II primed Foxp3-transduced T cells could markedly ameliorate CIA inflammatory responses at clinical (p<0.0014) and pathological exchanges including cellular infiltration (p=0.002), bone erosion (p=0.0013) and synovial hyperplasia (p=0.002). In contrast, collagen II unprimed Foxp3-transduced T cells like as collagen II primed or unprimed GFP-transduced T cells did not reveal any beneficial effects on arthritis features as compared with untreated group (p>0.05). Therefore, we believe that collagen II primed Foxp3-transduced T cells are interacting locally and systemically with immune cells which reveled with decreasing of T cells infiltration into joints along with specific CII IgG production. Considering the results described here, it appears that the using patients' T cells which previously exposed to specific antigens may have more effective therapeutic advantage in the production of induced regulatory T cells in the treatment of arthritis.

Background: Recent studies have revealed that multiple sclerosis (MS) lesions have distinct microRNA (miRNA) expression profiles. miR-181 family members show altered expression in MS tissues although their participation in MS pathogenesis remains uncertain. Herein, we investigated the involvement of miR-181a and miR-181b in the pathogenesis of MS and its animal model, experimental autoimmune encephalomyelitis (EAE)., Methods: miR-181a and -b levels were measured in the central nervous system (CNS) of patients with MS and mice with EAE as well as relevant leukocyte cultures by real-time RT-PCR. To examine the role of the miRNAs in leukocyte differentiation and function, miR-181a and -b mimic sequences were transfected into cultured primary macrophages and purified CD4 + T cells which were then analyzed by RT-PCR and flow cytometry. Luciferase reporter assays were performed to investigate the interaction of miR-181a and -b with the 3'-UTR of potential target transcripts, and the expression of target genes was measured in the CNS of EAE mice, activated lymphocytes, and macrophages., Results: Expression analyses revealed a significant decrease in miR-181a and -b levels in brain white matter from MS patients as well as in spinal cords of EAE mice during the acute and chronic phases of disease. Suppression of miR-181a was observed following antigen-specific or polyclonal activation of lymphocytes as well as in macrophages following LPS treatment. Overexpression of miR-181a and -b mimic sequences reduced proinflammatory gene expression in macrophages and polarization toward M1 phenotype. miR-181a and -b mimic sequences inhibited Th1 generation in CD4 + T cells and miR-181a mimic sequences also promoted Treg differentiation. Luciferase assays revealed Suppressor of mothers against decapentaplegic 7 (Smad7), as a direct target of miR-181a and -b., Conclusion: Our data highlight the anti-inflammatory actions of miR-181a and -b in the context of autoimmune neuroinflammation. miR-181a and -b influence differentiation of T helper cell and activation of macrophages, providing potential therapeutic options for controlling inflammation in MS.

MicroRNAs are small noncoding RNAs, which regulate the expression of protein coding transcripts through mRNA degradation or translational inhibition. Numerous reports have highlighted the role of miRNAs in regulating cell death pathways including the expression of genes involved in the induction of apoptosis. Tumor necrosis factor alpha (TNF-α) is a proinflammatory cytokine which can send pro-death signals through its receptor TNFR1. Diverse adaptor molecules including DENN/MADD adaptor protein have been shown to modulate TNF-α pro-death signaling via recruitment of MAP kinases to TNFR1 and activation of pro-survival NFκB signaling. Herein, we investigated the role of microRNA-181 (miR-181) in regulating DENN/MADD expression levels and its subsequent effects on TNF-α-induced cell death. Using bioinformatics analyses followed by luciferase reporter assays we showed that miR-181 interacts with the 3' UTR of DENN/MADD transcripts. miR-181 overexpression also led to decreased endogenous DENN/MADD mRNA levels in L929 murine fibroblasts. Flow cytometric analysis of miR-181 transfected cells showed this miRNA accentuates mitochondrial membrane potential loss caused by TNF-α. These findings were associated with enhanced apoptosis of L929 cells following TNF-α treatment. Overall, these data point to the potential role of miR-181 in regulating TNF-α pro-death signaling, which could be of importance from pathogenesis and therapeutic perspectives in inflammatory disorders associated with tissue degeneration and cell death.

Background: MicroRNAs have emerged as an important class of modulators of gene expression. These molecules influence protein synthesis through translational repression or degradation of mRNA transcripts. Herein, we investigated the potential role of miR-142a isoforms, miR-142a-3p and miR-142a-5p, in the context of autoimmune neuroinflammation., Methods: The expression levels of two mature isoforms of miR-142 were measured in the brains of patients with multiple sclerosis (MS) and the CNS tissues from mice with experimental autoimmune encephalomyelitis (EAE), an animal model of MS. Expression analyses were also performed in mitogen and antigen-stimulated splenocytes, as well as macrophages and astrocytes using real-time RT-PCR. The role of the mature miRNAs was then investigated in T cell differentiation by transfection of CD4 + T cells, followed by flow cytometric analysis of intracellular cytokines. Luciferase assays using vectors containing the 3'UTR of predicted targets were performed to confirm the interaction of miRNA sequences with transcripts. Expression of targets were then analyzed in activated splenocytes and MS/EAE tissues., Results: Expression of miR-142-5p was significantly increased in the frontal white matter from MS patients compared with white matter from non-MS controls. Likewise, expression levels of miR-142a-5p and miR-142a-3p showed significant upregulation in the spinal cords of EAE mice at days 15 and 25 post disease induction. Splenocytes stimulated with myelin oligodendrocyte glycoprotein (MOG) peptide or anti-CD3/anti-CD28 antibodies showed upregulation of miR-142a-5p and miR-142a-3p isoforms, whereas stimulated bone marrow-derived macrophages and primary astrocytes did not show any significant changes in miRNA expression levels. miR-142a-5p overexpression in activated lymphocytes shifted the pattern of T cell differentiation towards Th1 cells. Luciferase assays revealed SOCS1 and TGFBR1 as direct targets of miR-142a-5p and miR-142a-3p, respectively, and overexpression of miRNA mimic sequences suppressed the expression of these target transcripts in lymphocytes. SOCS1 levels were also diminished in MS white matter and EAE spinal cords., Conclusions: Our findings suggest that increased expression of miR-142 isoforms might be involved in the pathogenesis of autoimmune neuroinflammation by influencing T cell differentiation, and this effect could be mediated by interaction of miR-142 isoforms with SOCS1 and TGFBR-1 transcripts.

Maternal testosterone has been indicated to affect sex ratio of offspring. The present study was conducted to elucidate the role of androgen receptor in this regard by blockade of androgen receptor using flutamide in female mice. Mice were randomly assigned to two experimental groups. Mice in the control (n = 20) and treatment (n = 20) groups received 8 IU equine chorionic gonadotropin (eCG) followed by human chorionic gonadotropin (hCG) injection (8 IU) 47 hr later. In addition, mice in the control and treatment groups received four injections of ethanol-saline vehicle and flutamide solution (2.50 mg), respectively, started from 1 hr before eCG injection until hCG injection at 12-hr intervals. Conception rate was not different between the treatment (18/20: 90.00%) and control (19/20: 95.00%) groups (p > 0.05). Litter size was higher in the treatment (8.22 ± 0.26) than control (7.21 ± 0.28) group (p < 0.05). Male sex ratio was lower in the flutamide-treated mice (67/148: 45.30%) as compared with the untreated ones (80/137: 58.40%; odds ratio = 1.69; p < 0.05). In conclusion, the results showed that androgen receptor blockade could skew sex ratio of offspring toward females implying that the effect of testosterone on sex ratio might be through binding to androgen receptor. In addition, the blockade of androgen receptor using flutamide appeared to enhance litter size.

Dissimilar distribution of male and female calves within left and right uterine horns has been observed in beef cows. A retrospective study was conducted to investigate the effect of side of pregnancy on secondary sex ratio in Holstein dairy cows. Data associated with sex of calves, side of pregnancy, sire, dam, parity number of dam, AI technician, season and year were retrieved from the database of a Holstein dairy farm. In total, data consisted of 6515 birth records from 3155 dams and 244 sires across years 2001-2010. Data were analyzed using logistic regression. There was no difference in proportion of male and female calves between left (52.9% and 47.1%, respectively) and right (53.2% and 46.8%, respectively) uterine horns (P>0.05). AI technician, year, season and parity of dam did not affect secondary sex ratio (P>0.05). Secondary sex ratio of left and right uterine horns, and consequently, overall secondary sex ratio (53.1%) were skewed toward males as compared with hypothetical secondary sex ratio of 50% (P<0.05). Incidence of right pregnancy (60.5%) was higher than hypothetical 50% incidence of right pregnancy. In conclusion, the present study revealed similar secondary sex ratio of calves between left and right uterine horns in Holstein dairy cows., (Copyright © 2013 Elsevier B.V. All rights reserved.)

Various factors including synchronization treatments have been reported to influence sex ratio of offspring in cattle. The present study was conducted to investigate the effect of treatment with Presynch-Ovsynch protocol on sex ratio of offspring in Holstein dairy cows. Healthy Holstein cows (N = 1102) were randomly assigned to Presynch-Ovsynch (N = 564) or control (N = 538) group by parity. Cows in Presynch-Ovsynch group received two administrations of PGF(2)α 14 days apart started at Days 23 to 27 postpartum. Twelve days after the second PGF(2)α treatment, Ovsynch protocol, consisting of an administration of GnRH followed by administration of PGF(2)α 7 days later and a final administration of GnRH given 48 hours after the third PGF(2)α treatment, began. Cows were subjected to fixed-time AI 24 hours after the last administration of GnRH. Cows in control group received no treatment and were inseminated 12 hours after standing estrus. Sex of calves conceived by the first service postpartum was determined after parturition and used for calculation of sex ratio. Parity, season, sire, and calving to conception interval were considered as covariates in statistical analysis. Sex ratio of calves in Presynch-Ovsynch group (1.64) was higher than that in control group (1.09; odds ratio = 1.51; P < 0.05). Moreover, male to female ratio was higher in cows conceived in summer, fall, and winter than in cows that conceived in spring (P < 0.05). In conclusion, cows treated with Presynch-Ovsynch protocol and inseminated 24 hours after the last GnRH administration of Ovsynch had a higher sex ratio than cows inseminated 12 hours after standing estrus., (Copyright © 2013 Elsevier Inc. All rights reserved.)

There are growing numbers of recombinant proteins that have been expressed in milk. Thus one can consider the placement of any gene of interest under the control of the regulatory elements of a milk protein gene in a dairy farm animal. Among the transgene introducing techniques, only nuclear transfer (NT) allows 100 % efficiency and bypasses the mosaicism associated with counterpart techniques. In this study, in an attempt to produce a transgenic goat carrying the human coagulation factor IX (hFIX) transgene, goat fetal fibroblasts were electroporated with a linearized marker-free construct in which the transgene was juxtaposed to β-casein promoter designed to secret the recombinant protein in goat milk. Two different lines of transfected cells were used as donors for NT to enucleated oocytes. Two transgenic goats were liveborn. DNA sequencing of the corresponding transgene locus confirmed authenticity of the cloning procedure and the complementary experiments on the whey demonstrated expression of human factor IX in the milk of transgenic goats. In conclusion, our study has provided the groundwork for a prosperous and promising approach for large-scale production and therapeutic application of hFIX expressed in transgenic goats.

The serum levels of four important and well characterized inflammatory chemokines including MCP-1/CCL2, RANTES/CCL5, IL-8/CXCL8 and Fractalkine/CX3CL1 were evaluated in sulfur mustard (SM) exposed Iranian population 20 years after exposure. In this historical cohort study 372 SM exposed participants from Sardasht, and 128 unexposed participants as controls were studied. The serum concentrations of chemokines were measured by a sandwich ELISA technique. The serum concentrations in the exposed comparing to the control group were 201.86 vs 180.60 pg/ml (p=0.002), for MCP-1/CCL2, 1182.6 vs 1393.1pg/ml (p=0.021) for RANTES/CCL5, 12.61 vs 15 pg/ml (p=0.002) for IL-8/CXCL8 and 0.696 vs 0.0648 (p=0.413) for Fractalkine/CX3CL1. In conclusion, elevated levels of MCP-1/CCL2 may suggest an anti inflammatory response and decreased levels of IL-8/CXCL8 and RANTES/CCL5 may represent a different pathophysiology and diverse molecular mechanisms involved in long term clinical manifestations of SM exposure. However, further prospect into their role in the pathogenesis of SM remains to be done.

Background: Fasting during the month of Ramadan is one of the essential religious practices of Muslims. The aim of this study was to evaluate opsonisation, phagocytosis, and nitroblue tetrazolium (NBT) reduction by white blood cells in normal, healthy, male subjects under non-fasting (before Ramadan) and fasting (after Ramadan) conditions., Methods: In this study, 13 Muslim men, aged 28-54 years, whose health was confirmed by health application forms, gave blood samples one week before the beginning of the holy month of Ramadan and during the last week of Ramadan. Blood samples were tested for neutrophil phagocytosis, serum opsonisation power, and NBT reduction., Results: Despite a decline in the neutrophil phagocytic index and serum opsonisation index, the percentage of neutrophils participating in phagocytosis increased with fasting. In addition, there was an increase in the percentage of neutrophils demonstrating NBT reduction. Although there was a decrease in opsonisation of the serum, the increased percentage of opsonisation compensated for this defect., Conclusion: This study demonstrates the beneficial effect of fasting during Ramadan on neutrophil phagocytic function.

Background: The aim of this study was to investigate the effects of Ramadan fasting on neutrophil's respiratory burst and circulating immune complex (CIC) level., Methods: The effects of Ramadan fasting on neutrophil's respiratory burst and CIC was studied in 21 normal young fasting Muslim individuals using standardized chemiluminescence and poly ethylene glycol methods respectively, the results obtained and statistically analysed., Results: It was shown that in 11 cases out of 21 (52%) both of the chemiluminescence (CL) activity and CIC levels measured before and after Ramadan fasting were in normal range in spite of a insignificant decrease or increase in CL activity or CIC level. Therefore, the changes of the immunological parameters were not significant and the levels remained in the range of normal. In four cases out of 21 (24%), the CL activity and CIC levels were higher than normal range measured just before Ramadan, however after month of Ramadan the CL activity and CIC level decreased reaching to the normal level of these parameters. In four cases out of 21 (24%) there were an increase in CL activity and CIC levels after Ramadan fasting., Conclusion: There were no significant changes of CL activity of circulating neutrophils and CIC levels comparing the results obtained before and after Ramadan. More over there was a good correlation between these two immunological parameters measured in the present study.

Background: Fasting during Ramadan represent one of the five pillars of the Islamic religion. So it is important to know the effect of this kind of fasting on people's life especially on their health. The purpose of this study was to show the effect of fasting during Ramadan on circulating immune complex (CIC) level and immune system., Methods: The CIC levels were measured before and after Ramadan by polyethylenglycole method. The blood samples were examined using quantitation chemiluminesence and circulating immune techniques. The results were analyzed using t-test and calculated coefficient correlation between CIC before and after Ramadan., Results: There was no significant difference between the CIC level before and after Ramadan., Conclusion: According to the results, fasting during Ramadan does not have bad effect on immune system of healthy people.