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The present study aimed to clarify the alternations in alpha-fetoprotein (AFP), phosphorus (Ph), calcium (Ca), and metabolic hormones (Growth hormones (GH), insulin, and thyroid gland hormones) in singleton, twin, and triplet pregnancy in goats. Animals were examined for pregnancy ultrasonographically during mid-pregnancy (6-14 weeks of gestation), and were classified conferring to litter size into singleton (n =55), twin (n=25) and triple (n=6) pregnancy, and non-pregnant status (n=64). Blood samples (5 ml) were collected before ultrasound examination and serum was separated for analysis of alpha-fetoprotein, phosphorus, calcium, and GH, insulin, T3, and T4. The current data showed that the fetal numbers markedly influenced AFP, Ph, Ca, GH, insulin, T3 levels, but did not affect T4 and Ph/Ca ratio. AFP levels were higher in twin (P< 0.05) and triple (P< 0.001) bearing does than in non-pregnant does (56.00±5.00 and 87.00±1.90 vs 3.80±.051 ng/ml). Phosphorus and Ca levels declined with pregnancy, with the substantial decreased in multiple feti compared with non-pregnant does. Growth hormone imminently raised in twine and triple fetuses than non-pregnant does. Insulin levels elevated in triple compared to non- pregnant and singleton (P< 0.005). Serum free T3 levels were lower in single and twin bearing does than non-pregnancy. In conclusion, fetal number markedly influenced the dam metabolic profile and mineral homostatis through altering AFP and metabolic hormones levels during pregnancy in goats.

Obstructive sleep apnea (OSA) is associated with polycystic ovarian syndrome (PCOS), a common cause of infertility. Understanding predictors and outcomes of OSA in women with infertility may guide treatment.A descriptive cross-sectional survey was performed to assess OSA in women presenting to an infertility clinic using validated sleep questionnaires to assess sleep and fertility outcomes. An Infertile-C group (controls with male or tubal factors) and an Infertile-S group (unknown/other infertile causes) were analyzed to assess OSA risk and other sleep disorders (e.g., restless legs syndrome (RLS) and insomnia) with fertility outcomes (time to pregnancy, PCOS, irregular menstruation, and miscarriage).In 258 women, occurrences of OSA diagnosis (6%) and RLS (10%) were reported similar to women of child-bearing age in the general population. PCOS was unassociated with OSA risk. Predictors of OSA risk were BMI, insomnia symptoms, and sleep aid use. Obese women with high OSA risk were more likely to have other comorbidities (e.g., depression). In adjusted models, prior clinical OSA diagnosis was associated with miscarriage (odds ratio: 6.17 (1.24, 30.62), p = 0.026). RLS was associated with irregular menstruation (odds ratio: 3.73 (1.21, 11.53), p = 0.022).Similar to other populations, women with infertility and OSA risk have more health comorbidities and higher BMI and may present with insomnia symptoms. While the data are limited, this study supports the potential associations of OSA and miscarriage. Further work is needed to evaluate OSA in female infertility.

Introduction Kleine Levin Syndrome (KLS), or recurrent hypersomnia, is a rare sleep disorder characterized by recurrent episodes of severe hyper-somnolence lasting several days to several weeks. Because the etiology is unknown, treatment is mostly supportive. Some pharmacologic agents have been attempted to treat symptoms during episodes with limited success. Report of case(s) Patient is a 17 year-old with a history attention-deficit hyperactivity disorder and generalized anxiety disorder who presented to the clinic due to recurrent episodes of severe hypersomnolence associated with derealization and cognitive dysfunction during all episodes, anxiety of being alone, and other symptoms in some episodes such as altered taste affecting appetite, and binge eating. The patient was diagnosed with KLS and tracked episodes which lasted 11-12 days (average) with recurrence every 10-17 days. Labs, 72 hour EEG, MRI and MRA were largely unremarkable. The family declined trial of Lithium. Pharmacogenetic testing showed significant gene-drug interaction with duloxetine requiring higher drug dosage for effects. After psychiatry started duloxetine, the patient experienced a 7 month period without episodes. Then episodes recurred for unclear reason while on duloxetine but with slightly less duration (6-12 days) and shorter time between episodes. Duloxetine was discontinued due to lack of mood effects. Clarithromycin was trialed next for use during episodes without changes. Episodes were tracked for several months with some predictability. Duloxetine was restarted a second time after several months, and the patient showed mild improvement in event duration down to 8-11 days for a few months. Time between episodes also shortened to 8-10 days. Anxiety was only slightly reduced between and during the episodes, though they still continued to struggle with derealization and ability to focus during episodes that effected school and work functioning. Interestingly, they also started to have more insomnia between episodes while on duloxetine. Conclusion KLS is a rare disorder with limited data on effective treatments. To date, this is the first report of a prolonged brief KLS remission with duloxetine. Norepinephrine and serotonin modulation may limit episode duration. More research is needed to determine whether duloxetine can modulate the effects of KLS. Support (if any) None

Introduction The pivotal phase 3 REST-ON trial (NCT02720744) evaluated the efficacy and safety of once-nightly sodium oxybate (ON-SXB; FT218), an investigational extended-release formulation for treatment of adults with narcolepsy. In REST-ON, ON-SXB met its 3 coprimary endpoints: improvement in mean sleep latency on the Maintenance of Wakefulness test, Clinical Global Impression-Improvement rating (% much/very much improved), and number of weekly cataplexy attacks at all doses tested (P< 0.001 vs placebo). The safety profile of ON-SXB was consistent with that of immediate-release (IR) SXB. The ongoing RESTORE trial (NCT04451668) is an open-label/switch study evaluating the safety and tolerability of ON-SXB. Methods Participants aged ≥16 years with narcolepsy type 1 or 2 who completed the REST-ON trial, were on stable-dose (≥1 month) IR oxybate, or were oxybate-naive are eligible for RESTORE. Initial doses were 4.5 g/night or equivalent/closest to the previous total IR oxybate dose/night for those switching; incremental adjustments (1.5 g/week; maximum dose, 9 g/night) were allowed. Safety data for participants receiving ≥1 dose of ON-SXB as of 01 July 2022 are reported here. Results This analysis includes interim data from 180 participants (REST-ON participants, n=15 [8.3%]; oxybate-naive, n=35 [19.4%]; switch, n=130 [72.2%]). Most participants are white (n=150 [83.3%]) and female (n=122 [67.8%]); mean age is 35 years [range, 16–84]). Most participants who reported an adverse event (AE; n=105 [58.3%]) had AEs that were mild (61.9%) or moderate (32.3%) in severity. Three participants had serious AEs (abscess, deep vein thrombosis, rib fracture and pneumothorax); all were deemed unrelated to ON-SXB and all 3 participants continued in the study. Adverse drug reactions (ADRs; ie, AEs related/possibly related to study drug) were reported by 76 (42.2%) participants with 6 (3.3%) participants discontinuing ON-SXB owing to ADRs. ADRs occurring in ≥3% of participants were nausea (11.7%), somnolence (6.7%), headache (5%), enuresis (5%), somnambulism (3.9%), dizziness (3.9%), tremor (3.9%), and vomiting (3.3%). Conclusion Interim data from the RESTORE study indicate that ON-SXB is generally well tolerated with a low rate of discontinuation owing to ADRs. If granted final FDA approval, ON-SXB will offer adults with narcolepsy a once-at-bedtime oxybate treatment option. Support (if any) Avadel Pharmaceuticals

This study aimed to (1) identify the status of reproductive tract of repeat-breeder heifers (RBH) ultrasonography, as well as (2) determinethe variations in steroid hormones, oxidative stress biomarkers, and cell-free RNA in serum.These culled animals from400 heifers at the breeding stationwere examined ultrasonography, and whole blood samples were collected. Theultrasonic diagnosed heifers as paraovarian cyst (POC, n=18) associated with genital pathological conditionswere compared with groups of control heifers at follicular(n=6), luteal (n=6), and early pregnant (n=10). The POC appeared as anechoic circumscribed masses located nearby, but separated from the ovary. Follicular number, corpus luteum size, serum cell-free RNA, and ovarian steroids were not affected by POC, although dominant follicle size tended to be lower (P= 0.069)in POC withCL absence (POC-CL). Heifers with POC-CLhad lower MDA levels than pregnant ones. Buffaloes that had POC in the presence of CL (POC+CL) were found to have higher SOD activity than luteal and pregnant cases. Total serum proteinslevels recorded lower valuesin POC+CL compared to that in follicular phase. There was a significant positive relationship between POCsize, and GPx activity and CL size, but negative correlations with dominate follicles size. The POC might interfere with ovarian function, via alterations in antioxidant levels.

Background: Cryptosporidium parvum is a protozoan parasite of medical and veterinary importance that causes neonatal diarrhea in many vertebrate hosts. In this study, we evaluated the efficacy of an affinity-purified antigen as a C. parvum vaccine candidate using ileal and liver tissues of experimentally infected neonatal mice by immunohistochemical profiling and immune scoring of CD4, CD8, Caspase-3, and nuclear factor kappa B (NF-κB). This vaccine was prepared from the C. parvum oocyst antigen using immunoaffinity chromatography with cyanogen bromide-activated Sepharose-4B beads. Thirty neonatal mice were divided into three groups (10 mice/group): (1) nonimmunized noninfected, (2) nonimmunized infected (using gastric tubes with a single dose of 1×105 of C. parvum oocysts in 250 µl PBS solution 1 h before a meal) and (3) immunized (twice with 40 µg/kg of purified C. parvum antigen at 2-week intervals and then infected with 1×105 C. parvum oocysts simultaneously with the second group). After euthanizing the animals on the 10th day, post-infection, their ileal and liver tissues were collected and prepared for immunohistochemistry (IHC) staining to detect CD4, CD8, Caspase-3, and NF-κB levels, which are indicators for T helper cells, cytotoxic T cells, apoptosis, and inflammation, respectively. Results: The IHC results showed that CD4, CD8, Caspase-3, and NF-κB antibodies varied significantly (P <0.001) in both organs in all the groups. We also recorded high CD4 levels and low CD8 antibodies in the nonimmunized noninfected mice tissues, while the opposite was observed in the nonimmunized infected mice tissues. In the immunized infected mice, the CD4 level was higher than CD8 in both organs. While the Caspase-3 levels were higher in the ileal tissue of nonimmunized infected than immunized infected mice ileal tissues, the reverse was seen in the liver tissues of both groups. Furthermore, NF-κB expression was higher in the liver tissues of nonimmunized infected mice than in immunized infected mice tissues. Therefore, the IHC results and immune-scoring program revealed a significant difference (P <0.001) in the CD4, CD8, Caspase-3, and NF-κB antibody levels in both ileal and liver tissues of all mice groups, which might be necessary for immunomodulation in these tissues. Conclusions: The improvement observed in the immunized infected mice suggests that this vaccine candidate might protect against cryptosporidiosis.

Apparently healthy mares with conception failure or embryonic loss may have subclinical endometritis (SE). Our objective was to document evidence of systemic immune responses against SE in mares. In apparently healthy (control) mares as well as those with chronic endometritis (ChE) or subacute suppurative endometritis (SSE), both considered classes of SE, serum concentrations of cytokines (IL-1β, IL-6, IL-10 and TNF-α), chemokines (IL-8), acute phase protein (SAA), and plasma concentrations of prostaglandins (PGF2α and PGE2) were measured using validated enzyme linked immunoassays (EIA). Mixed-breed mares of known reproductive history, were used. Based on an endometrial cytological examination, mares were allocated into the following groups: healthy (control), ChE, and SSE (12, 26 and 11 mares, respectively). Serum concentrations of IL-6, IL-8 and IL-10 and plasma PGF2α concentrations were increased (P

This study aimed at evaluating the litter size influence on fetal growth (marked by biparietal diameter), steroid hormones (estradiol and progesterone), oxidative stress markers [Total antioxidant capacity (TAC), superoxide dismutase (SOD), catalase (CAT), Glutathione peroxidase (GPx), and malondialdehyde (MDA)], total proteins, and serum-free RNA. Goats (n=150) were blood sampled and assessed ultrasonographically during the mid-stage of pregnancy (6th to 14th week) and were classified into non-pregnant (n=64), single (n= 55) twine (n= 25), and triple (n= 6) pregnancy according to a number of feti. The correlation coefficient of caprine fetal growth was R² = 0.9609, 0.9418, and, 0.928 in single, twin, and triple feti, respectively. The area under the curve of the fetal growth was 286.2, 282.1, and 263.4 for single, twin, and triple caprine fetuses. The mean reduction rate in fetal growth compared to singleton pregnancy was 1.65±1.03 and 8.32±2.41 % in twine and triple feti, respectively. Estradiol significantly (P< 0.05) decreased, while progesterone (P< 0.01) and serum-free RNA (P< 0.001) increased in pregnant animals compared to non-pregnant. TAC and MDA increased in multiple pregnancies compared to non-pregnancy in association with the decrease of SOD and catalase activities. GPx activity and total proteins substantially decreased in triple pregnancy than non-pregnancy. Cell-free RNA negatively correlated with estradiol, CAT, GPx, and total proteins, and positively correlated with P4, TAC, and MDA. In conclusion, litter size greatly impacted fetal growth, maternal steroids, and serum-free RNA, and preload to oxidative stressmediated health disorders in pregnant goats.

SummaryThis study aimed to compare the expression of genes regulating follicles development, survival and steroid hormones secretion in oocytes and granulosa cells (GCs) and study the correlation between their expression and follicular fluid (FF) levels of progesterone (P4) in pregnant and non-pregnant camels. In total, 138 ovarian pairs from slaughtered camels were used. Gene expression and hormonal assay were determined using real-time polymerase chain reaction (PCR) and enzyme-linked immunosorbent assay (ELISA), respectively. The obtained results revealed that the number of follicles (3–8 mm) was significantly (P < 0.05) lower in pregnant, compared with non-pregnant, camels. P4 level in the FF was significantly (P < 0.05) higher in pregnant, compared with non-pregnant, camels. However, no significant (P > 0.05) difference was noticed in the oestradiol (E2) level. STAR, PTEN, IGF1 and BCL2 mRNA levels were significantly higher in GCs and significantly lower in oocytes of pregnant, compared with non-pregnant, camels. However, follicle-stimulating hormone receptor (FSHR) mRNA level was significantly lower in GCs and oocytes, and the BMP15 mRNA level was significantly lower in oocytes of pregnant, compared with non-pregnant, camels. P4 level in FF was positively correlated with STAR, PTEN, IGF1 and BCL2 mRNA levels in GCs and negatively correlated with BMP15 mRNA levels in oocytes and FSHR mRNA levels in GCs and oocytes of pregnant camels. It could be concluded that pregnancy-induced variations in oocytes and GC expression of BMP15, IGF1, FSHR, STAR, BCL2, and PTEN genes might be associated with a decrease in the number of follicles and an increase in the FF level of P4.

Fresh semen from fifty bulls was evaluated for ejaculate volume, individual motility, concentration, lives sperm and abnormalities as well as acrosome integrity. Bulls were classified according to semen motility into two groups; good and poor. DNA was extracted from semen of both groups, then the PCR followed by single-strand conformation polymorphism (SSCP) techniques were performed for mutation detection in gonadotropin releasing hormone receptor (GnRHR) gene through multiple sequence alignment. The results showed that the percentages of sperm motility, sperm concentration and live sperm, were significantly higher in good than poor semen quality bulls. However, semen volume, percentage of acrosome integrity and abnormalities did not differ between good and poor. The PCR amplification of 240-bp fragment and the results of SSCP appeared a genetic polymorphism with two patterns. Pattern I was seen in all good bulls, with incidence of 100%. Pattern II appeared only in poor semen quality bulls with a frequency of 31.25%. The sequence analysis of the PCR product for the two patterns showed two single nucleotide polymorphisms (SNPs) as a transversion base substitution mutation at positions 20 (T/A) and 193 (A/T). The GnRHR gene could be used as a genetic marker related to semen quality in buffalo due to the good semen bulls had a unique pattern.

Purpose The recent coronavirus disease (COVID-19) pandemic mainly affects the respiratory system; however, several oral and maxillofacial post-COVID-19 complications have also been observed. This series reports the growing number of osteonecrosis cases associated with post-COVID-19 patients. Materials and methods This is a retrospective, multi-center case series that reports cases with maxillary osteonecrosis after various periods of SARS-CoV-2 infection in the period between January and August 2021 based on the PROCESS guidelines. Results Twelve cases were reported with post-COVID-19 manifestation of spontaneous osteonecrosis of the maxillary jaw. Five patients were hospitalized during COVID-19 management and all of the twelve cases had at least one systematic Co-morbidity, and undertake corticosteroids prescription based on the COVID-19 disease treatment protocol. The mean onset of osteonecrosis symptoms appearance was 5.5 ± 2.43 weeks calculated from the day of the negative PCR test. The management was successfully done through surgical debridement and pre and post-operative antibiotics. No anti-fungal medications were prescribed as the fungal culture and the histopathological report were negative. Conclusion Post-COVID-related osteonecrosis of the jaw (PC-RONJ) could be now considered as one of the potential post-COVID-19 oral and maxillofacial complications that occurs unprovokedly and mainly in the maxilla.

This study was aimed to evaluate the yearly incidence of pediatric narcolepsy prior to and following the 2009 H1N1 pandemic and to evaluate seasonal patterns of narcolepsy onset and associations with H1N1 influenza infection in the United States. This was a multicenter retrospective study with prospective follow-up. Participants were recruited from members of the Pediatric Working Group of the Sleep Research Network including 22 sites across the United States. The main outcomes were monthly and yearly incident cases of childhood narcolepsy in the United States, and its relationship to historical H1N1 influenza data. A total of 950 participants were included in the analysis; 487 participants were male (51.3%). The mean age at onset of excessive daytime sleepiness (EDS) was 9.6 ± 3.9 years. Significant trend changes in pediatric narcolepsy incidence based on EDS onset (p

Supplementation of maturation media with antioxidant (bulk form) improves oocyte maturation. However, the influence of adding antioxidant (nano-particles) on oocyte maturation is not well known. We aimed to evaluate the effect of selenium nano-particles (SeNP) and bulk selenium (Se) on buffalo oocytes maturation, in terms nuclear maturation and molecular level. Oocytes were distributed into four groups; 1st group was control, 2nd group was supplied with Se (10 ng/ml), 3rd and 4th groups were supplied with 1 µg/ml SeNP (67 nm), and SeNP (40 nm), respectively. Matured oocytes were fixed and stained to determine nuclear maturation. Oocytes and COC after IVM were stored at − 80 °C, for RNA isolation and qRT-PCR for selected genes. The Se and seNP (40 nm) had a positive effect on oocytes nuclear maturation rates. Apoptosis-related cysteine peptidase (CASP3) was reduced in all supplemented groups. Anti-Mullerian hormone (AMH) was up-regulated in oocytes supplemented with SeNP (40 nm). In COC, AMH increased in group supplemented with SeNP (67 nm). In oocytes, phospholipase A2 group III (PLA2G3) decreased in all supplemented groups. While in COC, PLA2G3increased in group supplied with Se. In COC, luteinizing hormone/choriogonadotropin receptor (LHCGR) increased in groups supplied with Se or SeNP (40 nm).Glutathione peroxidase 4 (GPX4) increased in all supplemented groups, in oocytes and COC. In oocytes, superoxide dismutase (SOD) was up-regulated in supplemented groups {Se and SeNP (67 nm)}.The DNA methyltransferase (DNMT) in oocytes was reduced in supplemented groups. In oocytes, the POU class 5 homeobox 1 (OCT4) increased in all supplemented groups. In COC, the OCT4 was over-expressed in group supplemented with SeNP (40 nm). Selenium supplementation in bulk or nano-particle improved in vitro buffalo oocytes maturation, viaup-regulation of antioxidant defense and development competence genes. SeNP (smaller size, 40 nm) induced higher expression of antioxidant gene.

SummaryThis study aimed to: (i) characterize cultured granulosa cells (GCs) from different follicle sizes morphologically and molecularly; and (ii) select a suitable model according to follicular size that maintained GC function during culture. Buffalo ovaries were collected from a slaughterhouse and follicles were classified morphologically into: first group ≤ 4 mm, second group 5–8 mm, third group 9–15 mm and fourth group 16–20 mm diameter. GC pellets were divided into two portions. The first portion served as the control fresh pellet, and the secondwas used for 1 week for GC culture. Total RNA was isolated, and qRT-PCR was performed to test for follicle-stimulating hormone receptor (FSHR), cytochrome P450 19 (CYP19), luteinizing hormone/choriogonadotropin receptor (LHCGR), proliferating cell nuclear antigen (PCNA), apoptosis-related cysteine peptidase (CASP3), anti-Müllerian hormone (AMH), and phospholipase A2 group III (PLA2G3) mRNAs. Estradiol (E2) and progesterone (P4) levels in the culture supernatant and in follicular fluids were measured using enzyme-linked immunosorbent assay (ELISA). Basic DMEM-F12 medium maintained the morphological appearance of cultured GCs. The relative abundance of FSHR, CYP19, and LHCGR mRNAs was 0.001 ≤ P ≤ 0.01 and decreased at the end of culture compared with the fresh pellet. There was a fine balance between expression patterns of the proliferation marker gene (PCNA) and the proapoptotic marker gene (CASP3). AMH mRNA was significantly increased (P < 0.001) in cultured GCs from small follicles, while cultured GCs from other three categories (5–8 mm, 9–15 mm and 16–20 mm) showed a clear reduction (P < 0.001). Interestingly, the relative abundance of PLA2G3 mRNA was significantly (P < 0.001) increased in all cultured GCs. E2 and P4 concentrations were significantly (P < 0.001) decreased in all cultured groups. Primary cultured GCs from small follicles could be a good model for better understanding follicular development in Egyptian buffaloes.

Caregivers frequently report poor quality of life (QOL) in children with sleep-disordered breathing (SDB). Our objective is to assess the correlation between caregiver- and child-reported QOL in children with mild SDB and identify factors associated with differences between caregiver and child report.Analysis of baseline data from a multi-institutional randomized trial.Pediatric Adenotonsillectomy Trial for Snoring, where children with mild SDB (obstructive apnea-hypopnea index3) were randomized to observation or adenotonsillectomy.The Pediatric Quality of Life Inventory (PedsQL) assessed baseline global QOL in participating children 5 to 12 years old and their caregivers. Caregiver and child scores were compared. Multivariable regression assessed whether clinical factors were associated with differences between caregiver and child report.PedsQL scores were available for 309 families (mean child age, 7.0 years). The mean caregiver-reported PedsQL score was higher at 75.2 (indicating better QOL) than the mean child-reported score of 67.9 (Caregiver- and child-reported global QOL in children with SDB was weakly correlated, more so for young children. In pediatric SDB, child-perceived QOL may be poorer than that reported by caregivers. Further research is needed to assess whether similar trends exist for disease-specific QOL metrics.

Background No doubt that the corpus luteum (CL) plays a vital role in the regulation of female cyclicity in mammals. The scenarios among microRNAs (miRNAs) and their target genes and steroid hormones {estradiol (E2) and progesterone (P4)} are required for better understanding the molecular regulation of CL during its formation, maturation, and regression. We aimed to (I) study the changes in the relative abundance of miR-205, miR-26a-5p, miR-17-5p, and let-7b-5p and their target genes: LHCGR, CASP3, PCNA, AMH, and PLA2G3, during different stages of corpus luteum in Egyptian buffaloes, and (II) and to address different scenarios between steroid concentrations in the serum and the expression pattern of selected miRNAs and their targets. Methods The paired ovaries and blood samples were collected from apparently healthy 50 buffalo cows at a private abattoir. The ovaries bearing CL were macroscopically divided according to their morphological structure and color into hemorrhagic (CLH), developing (CLD), mature (CLM), regressed (CLR), and albicans (CLA). Small pieces from different stages of CL (CLH, CLD, CLM, CLR, and CLA) were cut and immediately kept at − 80 °C for total RNA isolation and qRT-PCR. The serum was separated for steroid level estimation. Results The LHCGR was expressed during different stages of CL, and the peak of expression was at the mid-luteal stage. The CASP3 revealed a stage-specific response at different stages of CL. The PCNA has an essential role in cellular proliferation in buffaloes CL. Both expression patterns of PLA2G3 and AMH were found over the various developmental and regression stages. It was noticed that miR-205 is conserved to target LHCGR and CASP3 transcripts. Moreover, CASP3 and AMH were targeted via miR-26a-5p. Additionally, the CASP3 and PLA2G3 were targeted via let-7b-5p. The P4 level reached its peak during CLM. There were positive and negative strong correlations between miRNAs (miR-26a-5p and miR-205), target genes (LHCGR and CASP3) during different stages of CL, and steroid hormones in the serum. Conclusions Taken together, the orchestrated pattern among miRNAs, target genes, and steroid hormones is essential for maintaining the proper development and function of CL in buffalo cows.

Background Extracellular vesicles (EVs) are a promising biomarker and play a vital role in cell–cell communication. This study aimed (I) to identify and characterize EVs from low volume uterine lavage (LVL) and serum in mares with endometritis, compared to healthy controls and (II) to measure serum levels of interleukin 6 (IL-6), and prostaglandins (PGF2α and PGE2). Mares were divided into 30 sub-fertile (endometritis) and 20 fertile (controls). Serum and LVL was collected for EV isolation, and determination of serum levels of inflammatory mediators. Characterization and visualization of EVs were done by electron microscopy, dynamic light scattering and flow cytometry. Results Serial ultracentrifugation of LVL and use of a commercial kit for serum were strategies for EVs isolation. Mares with endometritis released higher amounts of larger size EVs. The EVs from mares with endometritis differentially expressed CD9 and CD63, compared to controls. Mares suffering from endometritis evoked higher levels of inflammatory mediators. Conclusions Thus, EVs could be used for a better understanding the regulatory mechanisms associated with developing endometritis in mares.

A better understanding of the molecular mechanisms in granulosa cells (GC) is warranted, during different follicular and luteal developmental stages in buffalo cows. We aimed to (I) study the expression of selected genes in GC during follicular and luteal phases, (II) evaluate correlations between GC gene expression and steroid concentrations {17-beta estradiol (E2) and progesterone (P4)} in follicular fluid (FF), and (III) study effect of ovarian status on follicular population as well as follicular size frequency. Ovaries were collected in pairs from buffaloes (n = 178). Ovaries bearing corpus luteum (CL) were subdivided into hemorrhagic, developing, mature, and albicans. Follicles from luteal groups were classified only into small (

Introduction Restless Sleep Disorder (RSD) is characterized by frequent nocturnal movements of large muscle groups or a complaint of restless sleep by observers with associated daytime dysfunction.1 RSD, like other movement disorders of sleep, is associated with low iron stores.2 Children with RSD have increased NREM sleep instability and a propensity for parasomnia.2 No reports to date have demonstrated the use of iron therapy for parasomnia in patients with RSD. Furthermore, studies are limited in very young children with this disorder as they currently fall outside proposed diagnostic criteria.1 Report of Cases: A 2 year-old male presented to the sleep clinic with parental complaints of nightly sleep initiation and maintenance difficulty, mild snoring, restless sleep, episodes of sleepwalking and night terrors multiple times per week. Symptoms worsened over 6 months and were associated with daytime irritability and behavioral problems. Sleep quality measures assessed progress between visits: the PROMIS Sleep Disturbance (PROMIS SD) and Sleep Related Impairment (PROMIS SRI). Serum ferritin was initially 20ug/L. Polysmnogram demonstrated no significant sleep related breathing disorder, a large muscle group movement index of 14, periodic limb movement index of 3. Initial treatment included oral iron therapy, scheduled awakenings, and hypnotic dose melatonin. The sleep psychologist addressed limit setting and negative sleep associations with improved insomnia symptoms but parasomnia and restless sleep continued. The parent noted reduction in both movements and parasomnia frequency after one month of iron therapy. After a 3 month period on treatment, the parent reported complete cessation of the parasomnia, drastic reduction in restlessness, and near resolution of insomnia. Sleep quality measure PROMIS SD improved by 11, demonstrating reduced sleep disturbance. PROMIS SRI did not change. Ferritin level increased to 47ug/L. Conclusion RSD in a very young child with NREM parasomnia responded to oral iron therapy improving overall sleep quality. This case demonstrates the relationship between relative iron deficiency, RSD and associated parasomnia. Support (If Any) 1. DelRosso LM, Ferri R, Allen RP, et al. Consensus diagnostic criteria for a newly defined pediatric sleep disorder: restless sleep disorder(RSD).SleepMed.2020;75:335-340. 2. Leung W, Singh I, McWilliams S, Stockler S, Ipsiroglu OS. Iron deficiency and sleep - A scoping review. SleepMedRev.2020;51:101274

Cryptosporidium is an apicomplexan parasite of human and animals and is considered as an important co-factor in neonatal diarrhea. In this study, an explant culture was used as an in vitro model of buffalo intestine to evaluate the effect of Moringa leaves extract on Cryptosporidium parvum (C. parvum) oocysts using light and scanning electron microscopy and measuring IFN-γ, IL-12 and IL-14 in the culture supernatants. C. parvum oocysts were collected from naturally-infected calf feces, isolated, excysted and then co-inoculated with ileal tissue explants culture medium. The prepared Moringa leaves extract was then introduced to the infected tissues in the concentrations of 100 mg/ml and 300 mg/ml. After 24 h, tissues were collected and processed for light and scanning electron microscopy. Also, culture supernatants were collected for cytokines measurement. C. parvum parasitophorous vacuoles were found attached to the surface of tissue in Cryptosporidium-infected ileal tissue explants. High magnification imaging of ileal tissue explants using scanning electron microscopy showed that Moringa leaves extracts had a great effect on Cryptosporidium-infected ileal tissue explants. There was a high significant (P

Objective: To evaluate the teratogenic effects of clarithromycin on developing rabbit.Design: Randomized controlled study.Animals: Twenty one female New Zealand rabbits. Procedure: The pregnant female rabbits (21) were allocated into 4 groups. Control group (G1) (6 rabbits) received placebo. Other groups received clarithromycin (at a dose of 30 mg/kg b.wt orally) for ten days during the first stage (group2, n=6), the second stage (group3, n=6) or the third stage (group 4, n=3) of pregnancy. Three rabbits from each group were slaughtered on 20th and 30th days of pregnancy. Teratogenic effects were evaluated morphologically and histopathologically. Results: The therapeutic dose of clarithromycin exhibited morphological malformations, significant decrease (P≤0.05) in viable foeti mean, the mean values of the body weight and the fetal crown-rump length. However there was significant increase (P≤0.05) in dead foeti, fetal resorption rate, pre and post-implantation death in different stages of pregnancy. Visceral fetal abnormalities revealed dilatation of lateral ventricles of brain, thickening of the ventricular wall of the heart, and slight unilateral or bilateral widening of the renal pelvis. Skeleton fetal malformations showed incomplete ossification of 5th sternipri with absence of last ribs. Histopathological examination confirm teratogenic effects of clarithromycin. Conclusion and clinical relevance: The use of clarithromycin during pregnancy should be prohibited.

A better understanding of the molecular mechanisms in granulosa cells (GC) is warranted, during different follicular and luteal developmental stages in buffalo cows. We aimed to (I) study the expression of selected genes in GC during follicular and luteal phases, (II) evaluate correlations between GC gene expression and steroid concentrations {17-beta estradiol (E2) and progesterone (P4)} in follicular fluid (FF), and (III) study effect of ovarian status on follicular population as well as follicular size frequency. Ovaries were collected in pairs from buffaloes (n = 178). Ovaries bearing corpus luteum (CL) were subdivided into hemorrhagic, developing, mature, and albicans. Follicles from luteal groups were classified only into small ( 4 mm) and large (9-20 mm), while follicles from follicular groups were classified into three subgroups: small ( 4 mm), medium (5-8 mm), and large (9-20 mm). The FF and GC were collected for steroid concentrations measurement and gene expression, respectively. In the follicular phase, luteinizing hormone/choriogonadotropin receptor (LHCGR) and cytochrome P450 aromatase (CYP19) in small follicles decreased compared to medium ones. Large follicle showed an increase in LHCGR and CYP19 compared to medium ones. Follicle-stimulating hormone receptor (FSHR) decreased in large compared to medium size follicles. Proliferating cell nuclear antigen (PCNA) increased in small and large follicles. Meanwhile, anti-Mullerian hormone (AMH) and phospholipase A2 group III (PLA2G3) decreased in small and large follicles. The different stages of luteal phase had a profound impact on GC gene expression. There were strong (positive and/or negative) correlations between gene expression and steroid hormones. The different scenarios between expressed genes in GC and steroid concentrations are required for the proper growth and development of follicles and CL.

Background: So far the intimate link between serum microRNA (miRNA) and uterine inflammation in mares is unknown. We aimed (I) to investigate the expression profile of eca-miR-155, eca-miR-223, eca-miR-17, eca-miR-200a, and eca-miR-205 (II) and to measure the concentrations of interleukin 6 (IL-6), and prostaglandins (PGF2α& PGE2) in serum of Arabian mares with healthy and abnormal uterine status (endometritis).Methods and Results: This study was conducted on 80 Arabian mares; young (4-7 years), and old (8-14 years). These animals were divided into 48 sub-fertile including 16 young and 32 old mares suspected of endometritis and 32 fertile as control (24 young and 8 old) at stud farms. Serum samples were collected for measuring IL-6, PGF2α, and PGE2 concentrations, as well as serum miRNA isolation and qRT-PCR. Serum concentrations of IL-6, PGE2, and PGF2α were higher (P≤0.001) in mares with endometritis (young and old) compared to the control ones. Age of mares had a remarkable effect(0.001≤P≤0.01) onIL-6, PGE2, and PGF2αconcentrations. The relative abundance of eca-miR-155, eca-miR-223, eca-miR-17, eca-miR-200a, and eca-miR-205 was higher (P≤0.001) in both young and old mares with endometritis. We noticed that eca-miR-155, eca-miR-223, eca-miR-200a, and eca-miR-205 revealed higher (0.001≤P≤0.01) expression level in old than young mares with endometritis. Conclusions: To the best of our knowledge, this is the first study revealed that serum miRNA and serum inflammatory mediators (IL-6, PGE2, and PGF2α) could be used as non-invasive gold standard biomarkers, and therefore might be served as an important additional diagnostic tool for endometritis in Arabian mares.

We aimed (I) to simulate an in vivo milieu, through establishing an in vitro paradigm to study sperm-oviductal interactions using different segments of oviduct, as well as different incubation media, and (II) to investigate spatial changes of oviductal gene expression. Two experiments were designed; one was to investigate the yield of oviduct aggregates from different oviduct segments; in the second experiment, we observed effects of different incubation media on sperm-oviductal binding. Oviduct cell pellets before (control) and after sperm binding were collected for RNA isolation and gene expression. Isthmus resulted in a higher aggregate yield and possessed the highest affinity towards spermatozoa. The different segments of oviduct showed clear changes in gene expression after sperm binding. TALP medium promoted formation of a higher number of oviduct aggregates towards spermatozoa. Different media resulted in profound alterations in isthmus gene expression. Collectively, isthmus segment in TALP media showed the highest binding affinity to spermatozoa. At the molecular level, our in vitro model was successful for simulation in vivo milieu. Thus, our findings could be used as a simple tool to gain more insights into the molecular regulation of sperm movement, selection and affinity for oviductal binding in buffaloes.

We review the child-friendly and family-centered approaches to conducting pediatric sleep studies. While the majority of sleep laboratories are geared toward adults, there is a need and opportunity to offer pediatric services. Sleep laboratories that can accommodate children are better positioned to attract a broad range of new referrals. In this chapter, we discuss indications for pediatric polysomnography (PSG), implementation of a family-centered approach, how to prepare a sleep lab for children, engaging children and families with the PSG procedure, and pediatric-specific strategies for success and delivery of high-quality care. Experience and evidence-based recommendations about policies and procedures; equipment and sensors; atmosphere, staff training, and the importance of preparation are also included. With these tools and strategies for success, a sleep laboratory can provide “best practice” care for the pediatric patient and optimize patient experience.

Supplementation of maturation media with antioxidant (bulk form) improves oocyte maturation. However, the influence of adding antioxidant (nano-particles) on oocyte maturation is not well known. We aimed to evaluate the effect of selenium nano-particles (SeNP) and bulk selenium (Se) on buffalo oocytes maturation, in terms nuclear maturation and molecular level. Oocytes were distributed into four groups; 1st group was control, 2nd group was supplied with Se (10 ng/ml), 3rd and 4th groups were supplied with 1 µg/ml SeNP (67 nm), and SeNP (40 nm), respectively. Matured oocytes were fixed and stained to determine nuclear maturation. Oocytes and COC after IVM were stored at - 80 °C, for RNA isolation and qRT-PCR for selected genes. The Se and seNP (40 nm) had a positive effect on oocytes nuclear maturation rates. Apoptosis-related cysteine peptidase (CASP3) was reduced in all supplemented groups. Anti-Mullerian hormone (AMH) was up-regulated in oocytes supplemented with SeNP (40 nm). In COC, AMH increased in group supplemented with SeNP (67 nm). In oocytes, phospholipase A2 group III (PLA2G3) decreased in all supplemented groups. While in COC, PLA2G3increased in group supplied with Se. In COC, luteinizing hormone/choriogonadotropin receptor (LHCGR) increased in groups supplied with Se or SeNP (40 nm).Glutathione peroxidase 4 (GPX4) increased in all supplemented groups, in oocytes and COC. In oocytes, superoxide dismutase (SOD) was up-regulated in supplemented groups {Se and SeNP (67 nm)}.The DNA methyltransferase (DNMT) in oocytes was reduced in supplemented groups. In oocytes, the POU class 5 homeobox 1 (OCT4) increased in all supplemented groups. In COC, the OCT4 was over-expressed in group supplemented with SeNP (40 nm). Selenium supplementation in bulk or nano-particle improved in vitro buffalo oocytes maturation, viaup-regulation of antioxidant defense and development competence genes. SeNP (smaller size, 40 nm) induced higher expression of antioxidant gene.

To date, there is no an established protocol for total RNA isolation in Egyptian buffalo spermatozoa. The present study aimed (I) to establish a defined protocol for total RNA isolation from fresh and frozen spermatozoa, (II) to evaluate RNA quality and quantity from different extraction methods and studying gene expression. Warm and standard room temperature modified QIAzol Lysis Reagents were used for total RNA extraction. The quality and quantity of extracted RNA were checked, and subsequently qRT-PCR was performed using androgen receptor-like and three reference gene primers (GAPDH, ACTB and 18S). The warm modified QIAzol Lysis Reagents resulted higher yield of good quality RNA from fresh (569.54 ± 18.83 ng/μl) and frozen spermatozoa (110.59 ± 4.43 ng/μl), compared to standard room temperature modified QIAzol (421.26 ± 7.18 ng/μl) and (29.07 ± 5.25 ng/μl), for fresh and frozen semen samples respectively. The 260/280 ratio was 1.90 and 1.89 for fresh and frozen isolated semen by warm method respectively. The integrity of RNA was good and appeared as a sharp band on 2% agarose gel. The most stable reference gene was 18S. Reliable extraction method of high quality RNA yield could be a step forward for understanding mechanisms of spermatogenesis for improving male fertility.

This study aimed to: (i) characterize cultured granulosa cells (GCs) from different follicle sizes morphologically and molecularly; and (ii) select a suitable model according to follicular size that maintained GC function during culture. Buffalo ovaries were collected from a slaughterhouse and follicles were classified morphologically into: first group ≤ 4 mm, second group 5-8 mm, third group 9-15 mm and fourth group 16-20 mm diameter. GC pellets were divided into two portions. The first portion served as the control fresh pellet, and the secondwas used for 1 week for GC culture. Total RNA was isolated, and qRT-PCR was performed to test for follicle-stimulating hormone receptor (FSHR), cytochrome P450 19 (CYP19), luteinizing hormone/choriogonadotropin receptor (LHCGR), proliferating cell nuclear antigen (PCNA), apoptosis-related cysteine peptidase (CASP3), anti-Müllerian hormone (AMH), and phospholipase A2 group III (PLA2G3) mRNAs. Estradiol (E2) and progesterone (P4) levels in the culture supernatant and in follicular fluids were measured using enzyme-linked immunosorbent assay (ELISA). Basic DMEM-F12 medium maintained the morphological appearance of cultured GCs. The relative abundance of FSHR, CYP19, and LHCGR mRNAs was 0.001 ≤ P ≤ 0.01 and decreased at the end of culture compared with the fresh pellet. There was a fine balance between expression patterns of the proliferation marker gene (PCNA) and the proapoptotic marker gene (CASP3). AMH mRNA was significantly increased (P0.001) in cultured GCs from small follicles, while cultured GCs from other three categories (5-8 mm, 9-15 mm and 16-20 mm) showed a clear reduction (P0.001). Interestingly, the relative abundance of PLA2G3 mRNA was significantly (P0.001) increased in all cultured GCs. E2 and P4 concentrations were significantly (P0.001) decreased in all cultured groups. Primary cultured GCs from small follicles could be a good model for better understanding follicular development in Egyptian buffaloes.

Ameloblastic carcinoma (AC) is an extremely rare malignant odontogenic tumour arising from odontogenic epithelium. It was classified into primary type and secondary type. A previous study revealed that primary ameloblastic carcinoma cases were associated with more favourable prognosis than secondary cases. The aim of the present work was: to report the clinical, histopathological, immunohistochemical, and ploidy status, and therapeutic details of four cases of primary AC, and to review the literature with regard to clinical, follow-up, prognosis, histopathological, and immunohistochemical information of primary AC. The Medline database was searched using the term ameloblastic carcinoma and primary type. The review of English literature revealed that primary ameloblastic carcinoma favours the posterior mandible with profound male predilection and appears as an ill-defined radiolucency. Metastasis and invasion are more likely to occur in maxillary cases. The treatment of choice is wide surgical resection with or without cervical lymph node dissection. Adjuvant postoperative radiotherapy is beneficial in incomplete resection cases and advanced soft tissue invasion. The most specific diagnostic methods of AC, as concluded from review, are α-SMA in epithelial cells in conjunction with Ki-67 index value and SPF more than 11.5%.

A total of 138 pairs of slaughterhouse ovaries were collected from pregnant (n = 46) and non-pregnant (n = 92) she-camels. Oocytes (n = 150) and related granulosa cells (GCs) were retrieved from the growing follicles (3–8 mm) to study the effect of pregnancy status on the expression pattern of genes regulating follicles population and follicular fluid (FF) levels of estrogen (E2) and progesterone (P4). The mRNA fold changes of STAR, PTEN, and BCL2 genes were greater (p

This preliminary study investigated the tolerability and efficacy of a novel mattress technology-the Sound-To-Sleep (STS) system-in the treatment of sleep problems in children with autism.After screening, 45 children, ages 2.5 to 12.9 years, were randomized to order of mattress technology use (On-Off vs. Off-On). Treatment conditions (On vs. Off) lasted two weeks with immediate crossover. Tolerability, including study discontinuation and parent-report of mattress tolerance and ease of use, was tracked throughout the study. Efficacy assessments were obtained at baseline, prior to crossover, and end of study and included measures of autism traits, other psychopathology symptoms, sensory abnormalities, communication difficulties, quality of life, sleep diary parameters, and single-blinded actigraphy-derived sleep parameters. Statistical analyses evaluated differences in tolerability and efficacy when the STS system was on versus off.STS system use was well tolerated (n = 2, 4.4% dropout) and resulted in parent-reported sleep quality improvements (STS off mean = 4.3, 95% CI = 4.05-4.54 vs. on mean = 4.9, 95%CI = 4.67-5.14). The technology was described by parents as very easy to use and child tolerance was rated as good. Parent-diary outcomes indicated improvements in falling asleep and reduced daytime challenging behavior. Actigraphy-derived sleep parameters indicated improved sleep duration and sleep efficiency. Improvements in child and family quality of life were identified on parent questionnaires.A future large sample phase 2 trial of the STS system is warranted and would benefit from extended study duration, an objective primary efficacy outcome, and careful attention to methodological issues that promote compliance with the intervention and study procedures.

This chapter describes a case of alternating leg muscle activation (ALMA) followed by discussion of this condition and the related hypnagogic foot tremor (HFT). HFT and ALMA are benign sleep-related movements of unknown etiology, typically found incidentally on polysomnography. Both HFT and ALMA are characterized by bursts of tibialis anterior activity occurring in sequences with an inter-burst interval shorter than found in periodic limb movements in sleep. There is no specific treatment for HFT and ALMA. Rather, treatment should focus on the other comorbid sleep disorders such as obstructive sleep apnea, as HFT and ALMA motor activity associated with respiratory events may be responsive to positive airway pressure therapy. In this chapter the diagnostic criteria and clinical features are summarized, and polysomnographic tracings and video samples are provided.

This chapter illustrates the impact of sleep disorders in pregnancy by demonstrating a case of obstructive sleep apnea (OSA) associated with preeclampsia. Maternal-fetal health complications are well documented in association with maternal OSA. OSA in pregnancy is a risk for preeclampsia, eclampsia, gestational hypertension, and gestational diabetes mellitus. A growing body of literature supports the importance of detection and diagnosis of OSA in pregnant women. Pregnancy-related complications may be mitigated by OSA treatment. Treatment with continuous PAP therapy is well tolerated in pregnancy and can be offered during pregnancy. Further investigation is needed to understand the effects of treatment on outcomes in pregnancy.

The patient presented in this chapter had uncontrolled obstructive sleep apnea (OSA) and was intolerant of positive airway pressure (PAP). Previous conservative and surgical interventions failed. The patient was successfully treated with maxillomandibular advancement (MMA) surgery. Although many patients benefit from first-line therapy with PAP for OSA, some are intolerant of PAP and require discussion of alternative treatments. Maxillofacial surgical approaches are an option for the treatment of OSA in patients whose craniofacial anatomy is anatomically appropriate. With MMA, patients can achieve significant improvement and in many cases can achieve cure of OSA. Multiple studies support the benefits of MMA in those who are candidates for this treatment.

Cryptosporidium is an apicomplexan parasite of human and animals and is considered as an important co-factor in neonatal diarrhea. In this study, an explant culture was used as an in vitro model of buffalo intestine to evaluate the effect of Moringa leaves extract on Cryptosporidium parvum (C. parvum) oocysts using light and scanning electron microscopy and measuring IFN-γ, IL-12 and IL-14 in the culture supernatants. C. parvum oocysts were collected from naturally-infected calf feces, isolated, excysted and then co-inoculated with ileal tissue explants culture medium. The prepared Moringa leaves extract was then introduced to the infected tissues in the concentrations of 100 mg/ml and 300 mg/ml. After 24 h, tissues were collected and processed for light and scanning electron microscopy. Also, culture supernatants were collected for cytokines measurement. C. parvum parasitophorous vacuoles were found attached to the surface of tissue in Cryptosporidium-infected ileal tissue explants. High magnification imaging of ileal tissue explants using scanning electron microscopy showed that Moringa leaves extracts had a great effect on Cryptosporidium-infected ileal tissue explants. There was a high significant (P

Introduction Narcolepsy is a central disorder of hypersomnia with a bimodal peak incidence at ages 15 and 35. Onset in early childhood has been reported, but rarely under the age of 5. We present a case of Narcolepsy Type 1 in a 2.5-year-old with dropping spells. Report of case(s) A 2.5-year-old African American healthy male presented with acute onset sudden drops, unsteady gait, and staring spells. These episodes lasted a few seconds, occurred multiple times a day, and were associated with eye fluttering and dysarthric speech. History was notable for increased daytime sleep/sleep attacks, disrupted nocturnal sleep with confusional arousals, nightmares and sleepwalking, weight gain and mood instability with aggressive behavior. No preceding head injury or illness. Physical examination demonstrated frequent atonic episodes with loss of deep tendon reflexes. A clinical suspicion of myoclonic/atonic epilepsy prompted video- electroencephalogram (EEG) which showed frequent bursts of generalized intermittent slowing representative of NREM 1 sleep lasting several seconds. Magnetic resonance imaging of the brain and infectious workup was unremarkable. Cerebrospinal fluid (CSF) Orexin was Conclusion This case demonstrates an unusual presentation of Narcolepsy Type 1 in a toddler initially thought to have epilepsy. Video-EEG and low CSF orexin led to the diagnosis. His age, severity of cataplexy, mood changes, and parental concerns made treatment challenging. Furthermore, this case highlights the importance of early recognition of Narcolepsy in young children to provide appropriate treatment during critical developmental stages. Support (if any)

Maize is the one of important crops in Egypt. Aflatoxins (AFs) are the foremost cancer present compounds by Aspergillus flavus (A. flavus) and cause health risks to human and animals. This study aimed to suppression of aflatoxins production by A. flavus by using different concentrations of natural substances (carnation oil, lemongrass oil, propolis and beewax) and chemical substances (salicylic acid and potassium sorbate) on maize grains. The strains of A. flavus were isolated from local maize grains on Potato Dextrose Agar (PDA) and detect its ability of aflatoxins production on coconut agar media. Samples (100g) of sterilized maize grains were treated individually with different concentration of carnation oil, lemongrass oil, salicylic acid and potassium sorbate (0.25, 0.5, 1.0, 2.0, 4.0 and 6.0%) and at concentration 1% and 4% for propolis and beeswax each, then inoculated with A. flavus and stored for 30 days at 28±2˚C. AFs reduction was determined by using High Performance liquid Chromatography (HPLC). All the tested substances had active effect in inhibition of AFs production by A. flavus in stored maize grains. The production of Aflatoxin B1 and B2 (AFB1 and AFB2) was decreased to about 93% and 99% at concentration of 0.25% carnation oil. Lemongrass oil almost completely inhibited AFB1and AFB2 production (99.12% - 99.98% and 99.98% -99.99%, respectively) at concentration of 2% - 6%. Potassium sorbate and salicylic acid (0.25%- 6%) that were significantly effective controlling aflatoxins production on maize grains compared with control. While, the propolis and beewax found to be the most active to protect maize grains against fungi. Natural substances, carnation oil, lemongrass oil, beewax and propolis had higher active effect at low concentration on aflatoxin production more than salicylic acid and more safe for human used.

Bacterial infections of the genital tract are the major cause of reproductive failure in the mares. MiRNAs are important regulators of gene expression, mostly through transcriptional and translational regression. We hypothesized that LPS induced aberrant expression of miRNAs and their targets, which are involved in regulation of uterine homeostasis. Three groups of primary endometrial epithelial and stromal cells, and endometrial tissue explants were cultured. The 1st group was kept as control, while the 2nd and 3rd groups were challenged with low (0.5 μg/mL) or high (3.0 μg/mL) doses of Lipopolysaccharides (LPS). Cell pellets and tissue explants were collected after 24 and 48 h, for total RNA isolation and qRT-PCR of the selected miRNAs and their targets. Culture media and cell lysates were collected after 24 and 48 h, for cytokines (IL6 and TNFα) and prostaglandins (PGE2 & PGFα2) measurement. Both endometrial cells expressed TLR4 and its accessory molecules (MyD88 & CD14) that are required for triggering inflammatory immune response after LPS, via up-regulation of TRAF6, TNFα, IL6 and IL8, compared to the respective control. After both doses of LPS challenge, miR-155, miR-223 and miR-17 were significantly increased; miR-181b, miR-21 and let-7a were significantly decreased compared to respective controls. Interestingly, miR-24 and miR-532-5p were clearly up-regulated after only the low LPS dose. TNFα, IL6 and PGs in culture media and from cell lysates revealed dose- and time-dependent patterns, after LPS. Results indicated that both epithelial and stromal cells have a primary role in innate immune response after LPS challenge, while this recognition occurred via TLR4 and its accessory molecules. Dysregulation of miRNAs and their targets expression after LPS might affect normal uterine function through perturbation of PG and cytokine secretion.