Chromatin condensation but not DNA integrity of pig sperm is greater in the sperm-rich fraction

Background Protamination and condensation of sperm chromatin as well as DNA integrity play an essential role during fertilization and embryo development.In some mammals,like pigs,ejaculates are emitted in three separate fractions:pre-sperm,sperm-rich(SRF)and post sperm-rich(PSRF).These fractions are known to vary in volume,sperm concentration and quality,as well as in the origin and composition of seminal plasma(SP),with differences being also observed within the SRF one.Yet,whether disparities in the DNA integrity and chromatin condensation and pro-tamination of their sperm exist has not been interrogated.Results This study determined chromatin protamination(Chromomycin A3 test,CMA_(3)),condensation(Dibromobi-mane test,DBB),and DNA integrity(Comet assay)in the pig sperm contained in the first 10 m L of the SRF(SRF-P1),the remaining portion of the sperm-rich fraction(SRF-P2),and the post sperm-rich fraction(PSRF).While chromatin protamination was found to be similar between the different ejaculate fractions(P>0.05),chromatin condensation was seen to be greater in SRF-P1 and SRF-P2 than in the PSRF(P=0.018 and P=0.004,respectively).Regarding DNA integrity,no differences between fractions were observed(P>0.05).As the SRF-P1 has the highest sperm concentra-tion and ejaculate fractions are known to differ in antioxidant composition,the oxidative stress index(OSi)in SP,calcu-lated as total oxidant activity divided by total antioxidant capacity,was tested and confirmed to be higher in the SRF-P1 than in SRF-P2 and PSRF(0.42±0.06 vs.0.23±0.09 and 0.08±0.00,respectively;P<0.01);this index,in addition,was observed to be correlated to the sperm concentration of each fraction(Rs=0.973;P<0.001).Conclusion While sperm DNA integrity was not found to differ between ejaculate fractions,SRF-P1 and SRF-P2 were observed to exhibit greater chromatin condensation than the PSRF.This could be related to the OSi of each fraction.

In‑depth proteome characterization of endometrium and extraembryonic membranes during implantation in pig

Background Proteome characterization of the porcine endometrium and extraembryonic membranes is important to understand mother-embryo cross-communication.In this study,the proteome of the endometrium and cho-rioallantoic membrane was characterized in pregnant sows(PS)during early gestation(d 18 and 24 of gestation)and in the endometrium of non-pregnant sows(NPS)during the same days using LC-MS/MS analysis.The UniProtKB database and ClueGO were used to obtain functional Gene Ontology annotations and biological and functional networks,respectively.Results Our analysis yielded 3,254 and 3,457 proteins identified in the endometrium of PS and NPS,respectively;of these,1,753 being common while 1,501 and 1,704 were exclusive to PS and NPS,respectively.In addition,we iden-tified 3,968 proteins in the extraembryonic membranes of PS.Further analyses of function revealed some proteins had relevance for the immune system process and biological adhesion in endometrium while the embryonic chorion displayed abundance of proteins related to cell adhesion and cytoskeletal organization,suggesting they dominated the moment of endometrial remodeling,implantation and adhesion of the lining epithelia.Data are available via Pro-teomeXchange with identifier PXD042565.Conclusion This is the first in-depth proteomic characterization of the endometrium and extraembryonic mem-branes during weeks 3 to 4 of gestation;data that contribute to the molecular understanding of the dynamic environ-ment during this critical period,associated with the majority of pregnancy losses.

Oxytocin in pig seminal plasma is positively related with in vivo fertility of inseminated sows

Background:Identification of relevant in vivo biomarkers for fertility remains a challenge for the livestock industry.Concentrations of the small peptide hormone oxytocin(OXT),involved in male reproductive function and present in the seminal plasma(SP)of several species could be a robust one.This study characterized concentrations of SPOXT in ejaculates from boars used in artificial insemination(AI)programs aiming to evaluate its relationship with sperm quality variables and in vivo fertility of their liquid-stored AI-semen.Seminal OXT concentrations(ng/mL)were measured in 169 ejaculates from 61 boars of the Duroc,Pietrain,Landrace and Large White breeds using a direct competitive immunoassay test based on AlphaLISA®technology.Ejaculate(ejaculate volume,sperm concentration,total sperm count)and sperm parameters(motility,viability,intracellular generation of reactive oxygen species,plasma membrane fluidity)were assessed at 0 h and 72 h in AI-semen samples stored at 17℃.In vivo fertility included only 18 Large White and Landrace boars whose AI-semen was used to inseminated>100 sows and evaluated both farrowing rate and litter size of 3,167 sows.Results:The results showed that SP-OXT differed between boars and between ejaculates within boar(P<0.05)but not between breeds(Duroc,Pietrain,Landrace and Large White).Ejaculates with higher SP-OXT concentration/mL(hierarchically grouped;P<0.001)had larger volume and came from younger boars(P<0.05).Ejaculates of boars showing positive farrowing rate deviation exhibited higher(P<0.05)SP-OXT concentration/mL than those with negative farrowing rate deviation.Conclusion:The SP concentrations of OXT are boar,ejaculate and age dependent,and positively related with ejaculate volume and farrowing rates of liquid-stored semen AI-doses.

New Technique for Medial Patellar Desmotomy in Cattle and Donkeys

The present technique is designed to transect the medial patellar ligament indicated for treatment of upward fixation of patella in cattle and donkeys. A medial patellar desmotomy was performed first on cadavers and then on 21 live cattle or donkeys. In live animals under local anesthesia, a sterile silk strand (USP 1) mounted on a curved needle is inserted through skin at the medial aspect of the medial patellar ligament to the subcutaneous fascia and directed beneath the ligament to emerge at opposite side. The silk is then sawed to cut the ligament. In contrast to other procedures, the desmotomy is less invasive as skin at the surgical site is not incised, pericapsular fat and joint capsules not invaded, and the ligament is fully transected in a one-step procedure. Minimal tissue invasiveness limited infection of surgical site, minimized bleeding, and decreased related postoperative consequences.

Investigations on the Significance of Inductable Cyclooxygenase in Atherogenesis in Cholesterol-Fed Rabbits

Atherosclerosis is the most prevalent disease in middle and later years of human life. Heart attacks and strokes are among the most frequent causes of death. New approaches in therapy are the use of NSAID’s (nonsteroidal antiphlogistical drugs) of the new generation (selective inhibitors of COX-2) and DHEA (Dehydroepiandrostenon). The key enzyme of prostaglandin synthesis, the COX-2 isoenzyme, is predominantly found in inflammatory tissue. Out of this results a possible importance of COX-2-inhibitors in prophylaxis of atherosclerosis. Our study intended to examine the significance of COX-2 and the COX-2-formed prostanoids, as well as the significance of COX-2-independent iso-prostanes on atherosclerosis. For that purpose we tested the effect of the selective COX-2-inhibitor Celecoxib on rabbits fed with cholesterol and compared this with the effect of the steroid hormone DHEA in 4 groups: healthy control, cholesterol-fed control, DHEA-group and Celecoxib group. In the test prostanoids, nitrate and nitrite were measured by gas chromatograpy/tandem-mass-spectrometry (GC-MS/MS) in 24-hours-collected urine. Additionally we measured cholesterol and triglycerides in plasma. The aortas of the examinated animals were measured optically using a planimetrical method. The measurement of prostanoids, isoprostanes, nitrate and nitrite showed considerable variations and particulary significant differences (p < 0.01) even in the initial values. By the treatment with Celecoxib the rate of atherosclerosis was reduced in a highly significant way in comparison to the cholesterol group and the DHEA-group. Consequently the test demonstrated a significant role of COX-2 in the development of atherogenesis.

Induced Monocytes-Derived HSCs (CD34+) with LPS Accelerated Homing Rat Bone Marrow-Mesenchymal Stem Cell (BM-MSCs, CD105) in Injured Pancreas

Investigating the function of combining induced rat monocytes-derived bone marrow-haemopoietic stem cell (rat BM-HSCs) with LPS and rat bone marrow-mesenchymal stem cell (rat BM-MSCs) was to analyze the acceleration of homing process mechanism in injured pancreas. Mononucleated stem cells were isolated from aspirated whole rat BM using ficoll and cultured in α-MEM complete growth medium in 10 cm petridish. After two days, adherent cells after washing twice in petridish were added α-MEM growth medium and then mesenchymal cells were characterized using CD105 marker in third passage and labeled PKH26. Then haemopoietic stem cells (HSCs) were isolated with magnetic beads CD34+ and differentiated in vitro, and then induced monocytes with LPS. Animal experiment used 28 male Wistar rats, and divided them into 4 groups. After transplantation combined, both cells between monocyte derived HSc (mHSCs) and rat BM-MSC were analyzed expression of pair box gen 4 (Pax4), pancreatic and duodenal homeobox (Pdx1), C-peptide using immunohistochemistry, then secretion of insulin and C-peptide analyzed using indirect ELISA. Results showed that the expressions of Pax4, Pdx1, C-peptide found in the surface membrane cell of pancreatic cell, and secreted C-peptide and insulin were shown significant (P < 0.05) in transplanted group 2, 3 and 4, but in group 3 were transplanted with combined cells more dominant than non-combined cells. Conclusions suggested that combining of induced monocytes-derived HSCs and rat BM-MSCs has accelerated homing MSCs into injured pancreatic tissue.

Degradation of 3D-printed magnesium phosphate ceramics in vitro and a prognosis on their bone regeneration potential

Regenerative bone implants promote new bone formation and ideally degrade simultaneously to osteogenesis.Although clinically established calcium phosphate bone grafts provide excellent osseointegration and osteoconductive efficacy,they are limited in terms of bioresorption.Magnesium phosphate(MP)based ceramics are a promising alternative,because they are biocompatible,mechanically extremely stable,and degrade much faster than calcium phosphates under physiological conditions.Bioresorption of an implant material can include both chemical dissolution as well as cellular resorption.We investigated the bioresorption of 3D powder printed struvite and newberyite based MP ceramics in vitro by a direct human osteoclast culture approach.The osteoclast response and cellular resorption was evaluated by means of fluorescence and TRAP staining,determination of osteoclast activities(CA II and TRAP),SEM imaging as well as by quantification of the ion release during cell culture.Furthermore,the bioactivity of the materials was investigated via SBF immersion,whereas hydroxyapatite precipitates were analyzed by SEM and EDX measurements.This bioactive coating was resorbed by osteoclasts.In contrast,only chemical dissolution contributed to bioresorption of MP,while no cellular resorption of the materials was observed.Based on our results,we expect an increased bone regeneration effect of MP compared to calcium phosphate based bone grafts and complete chemical degradation within a maximum of 1.5-3.1 years.

Corrigendum to“Degradation of 3D-printed magnesium phosphate ceramics in vitro and a prognosis on their bone regeneration potential”[Bioact.Mater.19(2023)376-391/22]

The authors regret the reversal of the order of first and last names in the original article.The correct order of the authors’first and last names are Eugen Gefel,Claus Moseke,Anna-Maria Schmitt,Niklas Dümmler,Philipp Stahlhut,Andrea Ewald,Andrea Meyer-Lindenberg,Elke Vorndran.

Dog sperm head morphometry： its diversity and .=volution

Dogs have been under strong artificial selection as a consequence of their relationship with man. Differences between breeds are evident that could be reflected in seminal characteristics. The present study was to evaluate differences in sperm head morphometry between seven well-defined breeds of dog： the British Bulldog, Chihuahua, German Shepherd, Labrador Retriever, Spanish Mastiff, Staffordshire Terrier, and Valencian Rat Hunting dog. Semen samples were obtained by masturbation and smears stained with Diff-Quik. Morphometric analysis （CASA-Morph） produced four size and four shape parameters. Length, Ellipticity, and Elongation showed higher differences between breeds. MANOVA revealed differences among all breeds. Considering the whole dataset, principal component analysis （PCA） showed that PC1 was related to head shape and PC2 to size. Procluster analysis showed the British Bulldog to be the most isolated breed, followed by the German Shepherd. The PCA breed by breed showed the Chihuahua, Labrador Retriever, Spanish Mastiff, and Staffordshire Terrier to have PC1 related to shape and PC2 to size, whereas the British Bulldog, Valencia Rat Hunting dog, and German Shepherd had PC1 related to size and PC2 to shape. The dendrogram for cluster groupings and the distance between them showed the British Bulldog to be separated from the rest of the breeds. Future work on dog semen must take into account the large differences in the breeds＇ sperm characteristics. The results provide a base for future work on phylogenetic and evolutionary studies of dogs, based on their seminal characteristics.

The Prevalence and Surgical Repair of Canine Medial Patellar Luxation in Southern Taiwan

Medial patella luxation(MPL)is most common orthopedic disease in toy and miniature breeds.The breed predisposition in the United States has been reported for the Boston Terrier,Chihuahua,Pomeranian,Miniature Poodle,and Yorkshire Terrier[1].In Japan,beside Pomeranian and toy poodle,the Japanese Chin and Shiba are mostly affected breeds[2].Multiple surgical techniques have been applied successfully to correct such disorders including

Effect of honey on oxidation, chlorination and nitration by purified equine myeloperoxidase

Objective: To evaluate the antioxidant effect of honey using two classical methods generally used, and for the first time to test the effect of honey on the oxidation, chlorination and nitration by purified equine myeloperoxidase (MPO). Methods: The antioxidant activity of three Algerian honey samples (nectar honey, mixed honey and honeydew honey) was evaluated by two classical methods, the ferric- reducing/antioxidant power (FRAP) assay and 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical-scavenging capacity. Results: Honeydew honey had the highest reducing power and DPPH radical-scavenging activity, whereas nectar honey showed the lowest reducing power and DPPH radical-scavenging activity. All honey samples showed a significant inhibitory effect on the chlorination activity of equine MPO, but honeydew honey was the weakest inhibitor. The three samples were poorly inhibitor on the MPO oxidation and nitration activities, except for nectar honey that exerted an inhibitory effect at the highest tested concentration of 10%. These later results seem to contradict those obtained with DPPH and FRAP. Conclusions: The antioxidant capacity of honey is mainly due to the phenolic compounds and flavonoids it contained. It has been suggested that MPO might be involved in the antioxidant, not pro-oxidant, activity of phenolic compounds.