Dynamics of heat shock proteins and heat shock factor expression during heat stress in daughter workers in pre-heat-treated (rapid heat hardening) Apis mellifera mother queens.

Honeybees have a limited capacity to control their body temperature when exposed to thermal changes in the atmosphere. Environmental changes, such as thermal climate change, have an adverse effect on honeybee survival. Insects can be pre-heat-treated (rapid heat hardening) with a mild heat stressor to induce Hsp gene expression and protect them from future stresses. Sixty accepted mother queen (MQ) larvae at the age of 7 days were selected and divided into two incubation treatment groups (n = 30). The control group (non-heat-treated mother queens, nH-T MQ) was maintained at 34.5 °C for 15 min and 70% relative humidity (RH), and the pre-heat-treated (pre-heat-treated mother queens, pH-T MQ) group was subjected to 41 °C for 15 min and 70% RH. To evaluate the effect of larval pre-heat-treatment on thermotolerance acquisition of daughter workers, about 500 workers were collected from brood combs of each treatment group. The worker bees in their cages were incubated in digital-controlled thermo-incubators for 1 h under each of the following temperatures: 35, 40, 45, 50, 55, and 60 °C. The expression of Hsp40, Hsp60, Hsp70, HSC70-3, HSC70-4, HSC70-5, Hsp83, and Hsp90 genes in both head and thorax were evaluated by relative quantitative real-time qPCR (RT-qPCR). In general, the pH-T MQ workers exhibited a higher ability to tolerate temperature than nH-T MQ workers under extreme conditions. Furthermore, we reported for the first time that pre-heat treatment of the mother queen's larvae alters the basal and dynamic expression of heat shock proteins (Hsp40, Hsp60, Hsp70, and Hsp90) and heat shock factor (HSF) during thermoneutral conditions and during heat stress of daughter workers, probably indicating a substantial improvement of honeybees' thermotolerance acquisition in arid and semi-arid regions, and improvement of honeybee longevity and management strategies.

Association of single-nucleotide polymorphisms in the ESR2 and FSHR genes with poor ovarian response in infertile Jordanian women.

OBJECTIVE: Poor ovarian response (POR) refers to a subnormal follicular response that leads to a decrease in the quality and quantity of the eggs retrieved after ovarian stimulation during assisted reproductive treatment (ART). The present study investigated the associations of multiple variants of the estrogen receptor 2 (ESR2) and follicle-stimulating hormone receptor (FSHR) genes with POR in infertile Jordanian women undergoing ART. METHODS: Four polymorphisms, namely ESR2 rs1256049, ESR2 rs4986938, FSHR rs6165, and FSHR rs6166, were investigated in 60 infertile Jordanian women undergoing ART (the case group) and 60 age-matched fertile women (the control group), with a mean age of 33.60±6.34 years. Single-nucleotide polymorphisms (SNPs) were detected by restriction fragment length polymorphism and then validated using Sanger sequencing. RESULTS: The p-value of the difference between the case and control groups regarding FSHR rs6166 was very close to 0.05 (p=0.054). However, no significant differences were observed between the two groups in terms of the other three SNPs, namely ESR2 rs1256049, ESR2 rs4986938, and FSHR rs6165 (p=0.561, p=0.433, and p=0.696, respectively). CONCLUSION: The association between FSHR rs6166 and POR was not statistically meaningful in the present study, but the near-significant result of this experiment suggests that statistical significance might be found in a future study with a larger number of patients.

A study on the role of FMR1 CGG trinucleotide repeats in Jordanian poor ovarian responders.

The expansion of trinucleotide CGG repeats in the promoter of fragile X mental retardation 1 (FMR1) gene is associated with fragile X and fragile X associated tremor/ataxia syndromes. While the expansion of CGG repeats has been associated with such neuro/psychiatric diseases, the contraction of CGG repeats has been recently suggested as an indication of ovarian dysfunction. This study aimed to evaluate a possible association of the short CGG repeats with poor ovarian responders (POR) and to test for a possible correlation between the CGG size and different known markers of the ovarian reserve, namely FSH, AMH, and the number of retrieved oocytes from Jordanian females. We found a significant difference between the CGG median allele size between the cases and the controls (p < 0.001), where poor ovarian responders had shorter CGG repeats compared to the healthy controls. Also, females with alleles <26 had twice the odds to be presented in the POR compared to the controls. However, we did not find a significant correlation between CGG sizes and the markers of ovarian reserve. We conclude that although low CGG repeats appear to be linked to POR, the clinical utility of FMR1 for predicting ovarian response needs further investigation.

Genetics of Female Infertility: Molecular Study of Newborn Ovary Homeobox Gene in Poor Ovarian Responders.

BACKGROUND: Newborn ovary homeobox (NOBOX) gene plays a critical role in the transcriptional regulation of oocyte-specific genes. Previous studies have demonstrated a pathogenic effect of NOBOX variants on premature ovarian insufficiency (POI) patients. Poor ovarian response (POR) is a risk factor for POI. Therefore, genetic variants in the NOBOX gene may also be studied as risk factors for POR development. AIMS: The aim of the study is to investigate the association between seven known NOBOX single-nucleotide polymorphisms (SNPs) and POR in Jordanian females. SETTINGS AND DESIGN: This was a case-control study of 60 females with POR for controlled ovarian hyperstimulation and 59 healthy females with no history of reproductive problems. Blood samples were collected from the participants and seven SNPs of NOBOX gene were screened. SUBJECTS AND METHODS: DNA was extracted from blood samples. Polymerase chain reaction with primers specific for seven known SNPs in NOBOX gene was used to amplify the specified region within the gene followed by Sanger sequencing. RESULTS: The seven SNPs investigated in this study, namely, rs77587352 (c.271G>T, p. Gly91Trp), rs7800847 (c.349C>T, p. Arg117Trp), rs193303102 (c.907C>T, p. Arg303X), rs193303103 (c.1025G>C, p. Ser342Thr), rs193303104 (c.1048G>T, p. Val350Leu), rs201947677 (c.1064G>A, p. Arg355His), and rs146227301 (c.1856C>T, p. Pro619Leu), only represent the wild-type allele in both females with POR and healthy participants. CONCLUSIONS: The results show that only monomorphic genotype of the NOBOX variants was found in Jordanian females studied.