Metabolomic profiling of embryo culture media in patients with repeated implantation failure during assisted reproductive technology cycles.

Objective: This study investigated the metabolic status of the spent culture media from embryos of patients with repeated implantation failure (RIF) undergoing in vitro fertilization-intracytoplasmic sperm injection cycles in comparison with the embryos from healthy fertile women. Methods: Metabolite levels in spent culture media were assessed and compared between embryos from RIF patients (n=35) and oocyte donors as controls (n=15). Protein levels of insulin-like growth factor 1 (IGF-1) were determined using Western blotting. Concentrations of glucose, pyruvate, and lactate were measured using spectrophotometry. Ionic colorimetric assay kits were utilized to analyze the concentrations of sodium, chloride, calcium, and magnesium ions. High-performance liquid chromatography was employed to measure the concentrations of glutamic acid, aspartic acid, methionine, phenylalanine, and histidine. Results: Glucose consumption and lactate secretion were higher in the control group than in the RIF group. The magnesium concentration was significantly higher in the control group than in the RIF group, but glutamic acid and aspartic acid concentrations were lower in the control group than in the RIF patients (p<0.05). The levels of IGF-1, sodium, calcium, chloride, methionine, histidine, and phenylalanine did not show statistically significant differences between the two groups. Conclusion: The metabolic profile of the culture medium of the embryos in the RIF group differed from that of the control group. These findings suggest potential factors that may affect implantation capacity in RIF patients and provide a new perspective on embryo selection.

Radiographic assessment of normal parameters of forelimb distal phalanx in Turkmen horses.

The Turkmen horse is one of the oldest breeds in the world with unique characteristics in agility, endurance and jump, being publicly acclaimed. Laminitis affects the limbs and decreases athletic performance. Radiographic evaluation is essential to confirm the sinking diagnosis in every breed of a horse suffering from laminitis. Since no information about distal phalanx radiographic measurements in this breed has been reported, the current study was designed and conducted. In the present study, 24 clinically sound registered Turkmen horses of both sexes (15 mares and nine stallions) were selected. Lateromedial radiographic views of both front distal phalanges were taken, focusing on the distal phalanx. After magnification correction, images were used to measure founder distances (FD) and other distal phalanx radiographic measurements using an image processing program. There was no significant difference between radiographic measurements of the left and right front feet. The FD had a significant positive correlation with weight. The horses under 3 years of age had a significantly greater U angle than the horses over the age of three. Also, palmarocortical length (PCL) in the Turkmen stallions was significantly higher than mares. In this investigation, PCL in horses under the age of three was significantly lower than horses over 3 years of age. The values determined in this study can be used as baseline data of front feet in Turkmen horses.

How to maintain and transport equine adipose tissue for isolating mesenchymal stem cells?

BACKGROUND: Adipose tissue (AT) is one of the most important mesenchymal stem cell (MSC) sources because of its high quantities, availability and ease of collection. After being collected samples, they should be transported to a laboratory for stem cell (SC) isolation, culture and expansion for future clinical application. Usually, laboratories are distant from animal husbandry centers; therefore, it is necessary to provide suitable conditions for adipose tissue transportation, such that adipose-derived MSCs are minimally affected. In the current study, the impact of tissue maintenance under different conditions on MSCs derived from these tissues was evaluated. We aimed at finding suitable and practical transportation methods in which ASCs go through the slightest changes. RESULTS: In the current study, after being collected, equine AT was randomized into eight groups: four samples were maintained in stem cell culture media at 25 οC and 4 οC for 6 and 12 hrs. as transportation via SC media groups. Three samples were frozen at three different temperatures (- 20, - 75 and - 196 οC) as cryopreserved groups; these samples were defrosted 1 week after cryopreservation. Fresh and unfrozen AT was evaluated as a control group. The tissue samples were then initiated into enzymatic digestion, isolation and the culturing of SCs. Cells at passage three were used to evaluate the ability to form colonies, proliferation rate, plotting of the cell growth curve, and viability rate. All experiments were performed in triplicate. Stem cell isolation was successful in all groups, although purification of SCs from the first series of cryopreservation at - 196 οC and two series of - 20 οC was unsuccessful. There was no significant difference between the surface area of colonies in all groups except for - 20 οC. The growth rate of transportation via stem cell media at 25 οC for 6 hrs. was similar to that of the control group. MTT analysis revealed a significant difference between 25 οC 12 hrs. Group and other experimental groups except for control, 4 οC 12 hrs. and - 196 οC group. CONCLUSION: Data have shown freezing at - 75 οC, transportation via stem cell media at 4 οC for 12 hrs. and 25 οC for 6 hrs. are acceptable tissue preservation and transportation methods due to minor effects on MSCs features.

Relationship between sperm parameters and clinical outcomes of Intra Uterine Insemination (IUI).

BACKGROUND: Intrauterine insemination (IUI) is a widely utilized method for treating the infertile couples. The aim of the present study was to determine the pregnancy and abortion rates after IUI and to examine the relationship of sperm parameters with these rates. METHODS: This retrospective study was performed on 911 infertile couples undergoing IUI treatment in Shahid Akbarabadi IVF Centre from May 2017 to May 2019. To evaluate the correlation of sperm parameters with the clinical pregnancy and abortion rates, odds ratio (OR) with 95% confidence intervals (CI) was calculated. RESULTS: In this study, the pregnancy rate following IUI was 15.7% (143/911), and among women who achieved pregnancy, the abortion rate was 42.0% (60/143). According to the multiple logistic regression analysis, none of the sperm parameters was associated with the pregnancy rate. Couples with either male or female factor infertility etiologies were more likely to get pregnant than those with unexplained infertility. Regarding the abortion rate, multiple logistic regression analysis revealed that normal sperm count was related to a lower abortion rate (adjusted OR=0.25, 95% CI=0.07-0.91). CONCLUSION: The present study did not reveal a significant relationship between none of the sperm parameters and pregnancy rate after IUI treatment. However, among women who got pregnant, continuation of the pregnancy was associated with the normal sperm count. Furthermore, analysis of all semen parameters together in comparison to one parameter alone might be more accurate to predict pregnancy or abortion. Further prospective cohort studies with a large number of couples are required.

The impact of irritable bowel syndrome on health-related quality of life in women with polycystic ovary syndrome.

BACKGROUND: The objectives of this study were to compare the prevalence and quality of life (QOL) of irritable bowel syndrome (IBS) in women with polycystic ovary syndrome (PCOS) compared with healthy women. METHODS: This was a case-control study of 201 women recruited at an infertility clinic in Iran. The control group were healthy women (n = 100) and the comparison group, women with PCOS (n = 101). Data were collected by clinical Rome III criteria to determine the IBS, Bristol scale for stool consistency and IBS QOL. RESULTS: The reporting of IBS symptoms were higher in PCOS (20.7%) than control group (11%) (P = 0.05). The IBS QOL score in the IBS + PCOS group was lower than other groups (IBS+ non PCOS, non IBS + PCOS, non IBS+ non PCOS; scores in food avoidance and worries about health domains were significant (P < 0.01). CONCLUSIONS: We conclude that having PCOS and an increased level of LH/FSH tends to cause IBS symptoms. IBS + PCOS women experience significant impaired quality of life scores particularly in relation to worries about health and food avoidance. These results offer further insights into IBS in PCOS women and their functional status and wellbeing.

Use of bacterial cellulose film modified by polypyrrole/TiO2-Ag nanocomposite for detecting and measuring the growth of pathogenic bacteria.

The aim of this study was to use of bacterial cellulose/polypyrrole/TiO2-Ag (BC/PPy/TiO2-Ag) nanocomposite film to detect and measure the growth of 5 pathogenic bacteria. For this purpose, at first, 13 BC/PPy/TiO2-Ag films were fabricated, then bacterial suspensions were prepared according to McFarland standard. The results showed that by increasing the bacterial concentration, the electrical resistance of sensors was decreased and there was a relation between bacterial concentration and bacterial type with electrical resistance change of sensors. The obtained data showed that the sensitivity of the sensors was increased with increasing the concentration of polypyrrole and TiO2-Ag. FT-IR and SEM tests were performed to investigate the interaction between nanoparticles and determine the size of nanoparticles. The BC/PPy/TiO2-Ag biosensors are portable and the response time of these sensors is very short for target analysis. Therefore, these sensors have the potential to improve biological safety as diagnostic tools.

Does myoinositol supplement improve sperm parameters and DNA integrity in patients with oligoasthenoteratozoospermia after the freezing-thawing process?

Sperm cryopreservation is a routine method in andrology and IVF laboratory. However, the sperm quality and its fertilizing capacity have been decreased during this process. The purpose of this experiment was to determine the role of myoinositol as a supplement in amelioration of total and progressive sperm motility, DNA fragmentation, total antioxidant capacity (TAC), reactive oxygen species (ROS), and lipid peroxidation after the freezing-thawing process on patients with oligoasthenoteratozoospermia (OAT) syndrome. Semen samples obtained from 40 patients were divided into two aliquots and freezed with simple and 2 mg/mL myoinositol (MYO) supplemented freezing media. All samples were thawed and assessed after one month. Semen parameters were analyzed in terms of the motility by CASA, the level of total ROS by fluorimetry, TAC and MDA by colorimetric assay and finally DNA fragmentation by TUNEL assay. Our results clearly showed that MYO could improve total (37.46 vs. 12.91, p < 0.001) and progressive motility (21.92 vs. 6.49, p < 0.001) in experimental group compared to control group. A higher TAC level was observed in the MYO treated group in comparison to control group (1.11 vs. 0.91, p = 0.05). While MYO supplementation could not be effective on ROS level, it reduced DNA fragmentation of sperm after freeze-thaw process (p = 0.01). Therefore, MYO could be a good supplement for sperm freezing to reduce the detrimental effects of freezing process especially on DNA integrity, which is an important factor in the success of ART, in OAT suffered patients.

The effect of Myo-inositol on sperm parameters and pregnancy rate in oligoasthenospermic men treated with IUI: A randomized clinical trial.

BACKGROUND: In about 40% of the couples, the cause of infertility problems is attributed to men because of low sperm production and disturbed motility of sperm. Pieces of evidence show that Myo-inositol has a potential role for the treatment of sperm morphology and male fertility. OBJECTIVE: This study aimed to determine the effect of Myo-inositol on the sperm parameters and fertility rate in patients with oligoasthenospermia treated by intrauterine insemination (IUI). MATERIALS AND METHODS: This study was a randomized clinical trial conducted on 37 patients with oligoasthenospermia treated by IUI during 2016-2017. In this study, the patients were randomly divided into two groups of oligoasthenospermia treated with (Case group) and without Myo-inositol (Control group). The case group received 0.5 ml of Myo-inositol with a concentration of 2 mg/ml and incubated at 37°C incubator for 2 hr, but the control group had no interventions. RESULTS: The results of this study showed that although there was no significant difference in sperm parameters including sperm motility and concentration before processing with Myo-inositol in the case group, but there was a significant increase in sperm motility during the treatment with Myo-inositol. The therapeutic effect of this method was confirmed on induction of pregnancy in 18% of the treated patients, in such a way that was about twice greater than those who did not receive the drug. CONCLUSION: According to the results of this study, the use of Myo-inositol is efficient enough to change sperm parameters to increase the chance of fertility.

Application of mesenchymal stem cells to enhance non-union bone fracture healing.

ECM components include a number of osteoinductive and osteoconductive factors, which are involved in bone fracture healing. In this study, a combination of adipose derived mesenchymal stem cells (Ad-MSCs), cancellous bone graft (CBG), and chitosan hydrogel (CHI) was applied to the non-union bone fracture and healing effects were evaluated for the first time. After creation of animal models with non-union fracture in rats, they were randomly classified into seven groups. Radiography at 0, 2, 4, and 8 weeks after surgery, indicated the positive effects of Ad-MSCs + CBG + CHI and Ad-MSCs + CBG in treatment of bone fractures as early as 2 weeks after the surgery. These data were confirmed with both biomechanical and histological studies. Gene expression analyses of Vegf and Bmp2 showed a positive effect of Ad-MSCs on vascularization and osteogenic differentiation in all groups receiving Ad-MSCs, as shown by real-time PCR. Immunofluorescence analysis and RT-PCR results indicated existence of human Ad-MSCs in the fractured region 8 weeks post-surgery. In conclusion, we suggest that application of Ad-MSCs, CBG, and CHI, could be a suitable combination for osteoinduction and osteoconduction to improve non-union bone fracture healing. Further investigations are required to determine the exact mechanisms involved in this process before moving to clinical studies. © 2018 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 107A: 301-311, 2019.

COP1 plays a prominent role in drought stress tolerance in Arabidopsis and Pea.

Constitutively photomorphogenic 1 (COP1) is an E3 ubiquitin ligase that has been studied extensively in the photomorphogenesis- and light-related processes in Arabidopsis. However, the possible role of COP1 in plant drought stress response remains unknown. Hence, in the present study, the stomatal behavior as one of the key elements in plant dehydration response was investigated in Arabidopsis cop1-4 and pea light-independent photomorphogenesis (lip1) mutants. We observed that water loss rate in the cop1-4 and lip1 detached leaves was significantly much faster than wild-type, resulting from failing to reduce the stomatal aperture by the mutants. But, interestingly, the cop1-4 and lip1 isolated leaves treated with abscisic acid (ABA) as well as cop1-4 and lip1 soil-grown under drought stress could close their stomata as wild-type. Hence, COP1 plays a fundamental role in the regulation of stomatal movements in response to dehydration and its function was conserved during evolution in both Arabidopsis and pea. Further evaluations showed the cop1-4 mutant was not significantly damaged from the oxidative stress derived from soil water limiting conditions when compared to wild-type. Similarly, the up-regulation level of several key stress-responsive genes was relatively lower in cop1-4 than wild-type. Therefore, COP1 might be considered as a potential key regulator of both short-and long-term dehydration response. Multiple stress-related cis-elements were also detected in the COP1 promoter region, which supported its up-regulation in response to drought, salt, and cold stresses. Besides, we figured out the constitutively open stomata of cop1-4 in darkness can be as a result of the reduced AtMYB61 expression.

In vitro study of matrix metalloproteinases 1, 2, 9, 13 and serum amyloid A mRNAs expression in equine fibroblast-like synoviocytes treated with doxycycline.

Application of synthetic matrix metalloproteinases (MMPs) inhibitors, such as doxycycline is one of the possible therapeutic options for osteoarthritis. However, little is known about the protective mechanism of doxycycline in equine models on MMPs inhibitors as well as on serum amyloid A (SAA) gene expression. This study investigated the effects of doxycycline on mRNA expression of MMP-1, MMP-2, MMP-9, MMP-13, and SAA of equine fibroblast-like synoviocytes (FLSs). The FLSs were established from synovial fluids of clinically normal metacarpophalangeal joints of 6 skeletally mature horses. The cells were treated with either 10 or 100 µg/mL of doxycycline for 48 h. The mRNA expression of MMP-1, MMP-2, MMP-9, MMP-13, and SAA were assessed using real-time polymerase chain reaction (PCR). Treatment with doxycycline resulted in significantly decreased mRNA expression of MMP-1 in FLSs at both concentrations (P = 0.001). No significant differences were detected among groups for MMP-2, MMP-9, and MMP-13 (P > 0.05). Only a tendency towards a decrease in mRNA expression level of SAA in the presence of doxycycline could be detected. Doxycycline inhibits MMP-1 gene expression at the transcript level. These findings indicate that doxycycline can protect the articular environment through inhibition of MMP-1 at transcript level.

L'application d'inhibiteurs synthétiques des métalloprotéinases de la matrice (MMP), telle que la doxycycline, est une des options thérapeutiques possibles pour l'ostéoarthrite. Toutefois, peu de choses sont connues sur le mécanisme protecteur de la doxycycline dans les modèles équins des inhibiteurs des MMP, de même que sur l'expression génique de l'amyloïde sérique A (SAA). La présente étude visait à déterminer les effets de la doxycycline sur l'expression de l'ARNm de MMP-1, MMP-2, MMP-9, MMP-13, et SAA des synoviocytes équins apparentés aux fibroblastes (FLS). Les FLS ont été établis à partir du liquide synovial provenant d'articulations métacarpo-phalangiennes cliniquement normales de six chevaux squelettiquement matures. Les cellules ont été traitées avec 10 ou 100 µg/mL de doxycycline pendant 48 h. L'expression d'ARNm de MMP-1, MMP-2, MMP-9, MMP-13, et SAA a été évaluée par réaction d'amplification en chaîne par la polymérase en temps réel. Le traitement avec la doxycycline a causé une diminution significative de l'expression de MMP-1 par les FLS et ce pour les deux concentrations (P = 0,001). Aucune différence significative ne fut détectée parmi les groupes MMP-2, MMP-9, et MMP-13 (P > 0,05). Seulement une tendance à la diminution de l'expression d'ARNm de SAA en présence de doxycycline pouvait être notée. La doxycycline inhibe l'expression génique de MMP-1 à l'étape de la transcription. Ces informations indiquent que la doxycycline peut protéger l'environnement articulaire en inhibant MMP-1 à l'étape de la transcription.(Traduit par Docteur Serge Messier).

Using of Ileucecal Interposition as a Neo-stomach in Dogs with Total Gastrectomy (Presentation of a Novel Technique).

Total gastrectomy and R&Y reconstruction is the standard treatment of gastric cancer except distal form one. Malnutrition is a common adverse effect of this technique, and in this study, we preset a novel technique in order to help to reduce the rate of it. We use seven dogs as participants and after total gastrectomy; we used the 10 cm of the ileum and its concomitant cecum as a neo-stomach in them. For 3 months, the dogs were evaluated in nutrition status. Three of the dogs died-one due to endocarditis and two due to internal hernia. No anastomosis leakage or stricture was seen and there were no unusual neo-stomach dilation or food stasis. This technique can help the nutrition status of total gastrectomy patients but needs more comparative human studies.

Intra- and Postoperative Complications of Lateral Maxillary Sinus Augmentation in Smokers vs Nonsmokers: A Systematic Review and Meta-Analysis.

PURPOSE: This meta-analysis and systematic review focused on the following question: Does tobacco smoking increase the risk of intra- or postoperative complications of lateral maxillary sinus floor elevation? MATERIALS AND METHODS: The following electronic databases were searched up to and including November 2015 without language restriction: CENTRAL, MEDLINE, Google Scholar, Scopus, Sirous, and Doaj. Studies were included if rates of intra-or postoperative complications of sinus floor elevation in smokers and nonsmokers were recorded separately. The following complications were assessed: sinus membrane perforation, bleeding, wound dehiscence, wound infection, sinusitis, hematoma, and oroantral fistula. The Critical Appraisal Skills Programme was used to assess the risk of bias in included studies. Random-effects meta-analyses were used to assess the number of each complication in smokers and nonsmokers. RESULTS: Out of 929 eligible publications, 11 articles were included. Meta-analysis of the studies revealed a significantly increased risk of developing wound dehiscence after sinus floor elevation among smokers compared with nonsmokers (Risk Ratio [RR]: 7.82; 95% confidence interval [CI]: 2.38, 25.74; P = .0007). Moreover, risk of developing wound infection was greater in smokers when prospective studies were included in the meta-analysis (RR: 5.33; 95% CI: 1.34, 21.25; P = .02). However, the meta-analysis of included studies did not show significant differences between smokers and nonsmokers concerning risk of sinus membrane perforation and bleeding during sinus floor elevation (P = .46 and P = .33, respectively). CONCLUSION: Considering the lack of randomized controlled trials and the small number of included studies, the results indicate that smoking seems to be associated with increased risk of wound dehiscence and infection after the sinus augmentation procedure.

A case report of partial bilateral hind limb adactyly in a male lamb.

Hemimelia as a congenital anomaly is a failure of development of extremities formation in embryonic period. This anomaly is defined as complete absence of the part of extremities and different forms were explained for hemimelia. Adactyly is an alternative name for transverse hemimelia and is a rare disorder in the most of animal species. A two months old male lamb with normal vital signs was referred to clinic due to both hind limbs shortness and absence of hooves from the birth day. Clinical and radiological examinations were performed and partial hemimelia was confirmed radiographically in both hind limbs. In left hind limb, total absence of the toe indicated presence of adactyly in this limb. No other congenital deformities were diagnosed in skeletal system based on clinical and radiological examinations. According to our knowledge, this is the first report of such rare conditions in a lamb. Clinical findings and radiological signs of this rare anomaly in a lamb were described in this report.