Seminal plasma contains a large protein component which has been implicated in the function, transit and survival of spermatozoa within the female reproductive tract. However, the identity of the ...majority of these proteins remains unknown and a direct comparison between the major domestic mammalian species has yet to be made. As such, the present study characterized and compared the seminal plasma proteomes of cattle, horse, sheep, pig, goat, camel and alpaca. GeLC–MS/MS and shotgun proteomic analysis by 2D–LC–MS/MS identified a total of 302 proteins in the seminal plasma of the chosen mammalian species. Nucleobindin 1 and RSVP14, a member of the BSP (binder of sperm protein) family, were identified in all species. Beta nerve growth factor (bNGF), previously identified as an ovulation inducing factor in alpacas and llamas, was identified in this study in alpaca and camel (induced ovulators), cattle, sheep and horse (spontaneous ovulators) seminal plasma. These findings indicate that while the mammalian species studied have common ancestry as ungulates, their seminal plasma is divergent in protein composition, which may explain variation in reproductive capacity and function. The identification of major specific proteins within seminal plasma facilitates future investigation of the role of each protein in mammalian reproduction.
This proteomic study is the first study to compare the protein composition of seminal plasma from seven mammalian species including two camelid species. Beta nerve growth factor, previously described as the ovulation inducing factor in camelids is shown to be the major protein in alpaca and camel seminal plasma and also present in small amounts in bull, ram, and horse seminal plasma.
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•Cross species comparison of the seminal plasma proteomes of seven mammalian species•The similarity of seminal proteomes can be related to the phylogeny of the species.•Beta nerve growth factor is the major protein in alpaca and camel seminal plasma.•Bodhesins and zinc alpha glycoprotein are a signature of small ruminant seminal plasma proteomes.•Nucleobindin-1 and RSVP14 proteins are common to the seven species.
Seminal plasma is composed of secretions from the epididymis and the accessory sex glands and plays a critical role in the fertilising ability of spermatozoa. In rams, analysis of seminal plasma by ...GeLC–MS/MS has allowed the identification of more than 700 proteins, including a high abundance of Binder of Sperm family proteins (BSP1, BSP5, SPADH1, SPADH2), the spermadhesin family (bodhesin2), lactoferrin and newly identified proteins like UPF0762 (C6orf58 gene). When spermatogenesis was stopped by scrotal insulation, changes in the proteome profile revealed the sperm origin of 40 seminal proteins, such as glycolysis pathway enzymes, the chaperonin containing TCP1 (CCT) complex and the 26S proteasome complex. Sperm mobility after liquid preservation (24h in milk at 15°C) is male dependent and can be correlated to differences in the seminal plasma proteome, detected by spectral counting. The negative association of zinc alpha-2 glycoprotein (ZAG) with semen preservation was confirmed by the use of recombinant human ZAG, which induced an increase in mobility of fresh sperm, but then decreased sperm mobility after 24h of incubation. Several sperm membrane proteins interacting with the cytoskeleton, glycolysis enzymes and sperm-associated proteins involved in capacitation correlated with better liquid storage and can be considered as seminal biomarkers of sperm preservation.
Extensive analysis of the ram seminal plasma proteome reveals a complex and diverse protein composition. This composition varies between males with different sperm preservation abilities. Several proteins were shown to originate from the spermatozoa and positively correlate with sperm liquid preservation, indicating that these proteins can be traced as sperm biomarkers within the seminal plasma. The zinc alpha-2 glycoprotein (ZAG) was found to have a biphasic effect on sperm mobility, with a short-term stimulation followed by a long-term exhaustion of sperm mobility after a 24h preservation period.
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•The seminal plasma proteome can be associated with the sperm preservation ability.•Several seminal proteins from sperm origin are biomarkers of sperm preservation.•The ZA2G increases mobility of fresh spermatozoa but decreases liquid preservation.
Calix2benzo4pyrrole m‐6 and p‐6, each containing two dipyrromethane moieties and two m‐phenylene or p‐phenylene units, respectively, were readily synthesised from pyrrole, 1,3‐ and ...1,4‐bis(1,1′‐dimethylhydroxymethyl)benzene, (m‐4 and p‐4, respectively) and acetone. Macrocycles m‐6 and p‐6 were tested as receptors for a selection of anions, such as acetate, dihydrogenphosphate and fluoride. The X‐ray structures of m‐6 and p‐6 and those of the complexes m‐6⋅F−, m‐6⋅Cl− and m‐6⋅CH3COO− (with an nBu4N+ counterion) were also determined.
Novel expanded and hybrid calixpyrroles 1 and 2 were synthesised and tested as hosts for anions. Compound 2 formed 1:1 inclusion complexes with fluoride, chloride, dihydrogenphosphate and acetate (X‐ray structure of 2⋅AcO− is shown here), and it was generally a better host than 1 and the smaller, related calix4pyrrole.
The structures of poly(1,4-phenylenesulfone), 1,4-ArSO2 n (Ar = 1,4-phenylene), and its tetrasulfone oligomer ArSO2ArSO2ArSO2ArSO2Ar (Ar = phenyl or 1,4-phenylene) have been determined from X-ray ...powder diffraction data interfaced to molecular simulation and diffraction modeling. An initial model for the tetrasulfone oligomer was developed using conformational and packing information obtained from a single-crystal X-ray determination of the structure of the trisulfone ArSO2ArSO2ArSO2Ar, in which the aromatic ring planes lie essentially orthogonal to a plane defined by the bridging C−S−C groups. Energy-minimization and crystallographic refinement of the tetrasulfone structure by Pawley and Rietveld methods yielded an agreement factor (R wp) of 15.3%., and an analogous approach led to solution of the crystal structure of poly(1,4-phenylenesulfone) (R wp = 5.5%). In all three structures, the diaryl sulfone units adopt near-perfect open-book conformations and the molecules crystallize with the aromatic rings of laterally adjacent chains essentially parallel. The polymer unit cell is C-centered orthorhombic (two chains per cell), space group Cmcm, with a = 10.79, b = 5.05, and c = 9.97 Å. The polymer chain adopts 21 helical geometry, and crystal packing is dominated by SO···H−C interactions (O···H = 2.68 Å).
Irinotecan and 5-fluorouracil (5-FU) are used to treat metastatic colorectal cancer. Irinotecan's active metabolite is inactivated by UDP-glucuronosyltransferase 1A1 (UGT1A1), which is deficient in ...Gilbert's syndrome. Irinotecan and metabolites are transported by P-glycoprotein, encoded by ABCB1. 5-FU targets folate metabolism through inhibition of thymidylate synthase (TYMS). Methylenetetrahydrofolate reductase (MTHFR) generates active folate necessary for haematopoiesis. We retrospectively genotyped 140 Swedish and Norwegian irinotecan and 5-FU-treated colorectal cancer patients from the Nordic VI clinical trial for selected variants of UGT1A1, ABCB1, TYMS and MTHFR. We found an increased risk of clinically relevant early toxicity in patients carrying the ABCB1 3435 T/T genotype, Odds ratio (OR)=3.79 (95% confidence interval (CI)=1.09-13.2), and in patients carrying the UGT1A1(*)28/(*)28 genotype, OR=4.43 (95% CI=1.30-15.2). Patients with UGT1A1(*)28/(*)28 had an especially high risk of neutropenia, OR=6.87 (95% CI=1.70-27.7). Patients who had reacted with toxicity during the first two cycles were in total treated with fewer cycles (P<0.001), and less often responded to treatment (P<0.001). Genetic variation in ABCB1 was associated with both early toxicity and lower response to treatment. Carriers of the ABCB1 1236T-2677T-3435T haplotype responded to treatment less frequently (43 vs 67%, P=0.027), and survived shorter time, OR=1.56 (95% CI=1.01-2.45).
Agranulocytosis is a serious, although rare, adverse reaction to sulfasalazine, which is used to treat inflammatory joint and bowel disease. We performed a genome‐wide association study comprising ...9,380,034 polymorphisms and 180 HLA alleles in 36 cases of sulfasalazine‐induced agranulocytosis and 5,170 population controls. Sulfasalazine‐induced agranulocytosis was significantly associated with the HLA region on chromosome 6. The top hit (rs9266634) was located close to HLA‐B, odds ratio (OR) 5.36 (95% confidence interval (CI) (2.97, 9.69) P = 2.55 × 10−8). We HLA‐sequenced a second cohort consisting of 40 cases and 142 treated controls, and confirmed significant associations with HLA‐B*08:01, OR = 2.25 (95% CI (1.02, 4.97) P = 0.0439), in particular the HLA‐B*08:01 haplotype HLA‐DQB1*02:01‐DRB1*03:01‐B*08:01‐C*07:01, OR = 3.79 (95% CI (1.63, 8.80) P = 0.0019), and with HLA‐A*31:01, OR = 4.81 (95% CI (1.52, 15.26) P = 0.0077). The number needed to test for HLA‐B*08:01 and HLA‐A*31:01 to avoid one case was estimated to be 1,500. We suggest that intensified monitoring or alternative treatment should be considered for known carriers of HLA‐B*08:01 or HLA‐A*31:01.
Calix6pyrrole 2 and the “hybrid systems” calix3furan3pyrrole 12, calix2furan4pyrrole 13, and calix1furan5pyrrole 14, have been synthesized by increasing conversion of the furan units present in the ...readily accessible calix6furan 3 to pyrroles. The host–guest chemistry of these novel macrocycles towards a number of anions, including halogen ions, dihydrogen phosphate, hydrogen sulfate, nitrate, and cyanide has been investigated in solution by 1H NMR titration techniques and/or phase transfer experiments. The solid‐state structures of the free receptors 2, 12, and 13, the 1:1 complexes of calix6pyrrole 2 with chloride and bromide, and the 1:1 complex of 14 with chloride are also reported.
A number of “hybrid” calixnfuranmpyrrole analogues (m + n=6) and calix6pyrrole (1) have been synthesized from calix6furan 2 (see picture). Their host–guest chemistry towards a number of anions (F−, Cl−, Br−, I−, H2PO4−, HSO4−, NO3−, and CN−) has been investigated in solution by NMR spectroscopy and, where possible, in the solid‐state by X‐ray crystallography. The association constants for many of the macrocycles with the screened anions have also been determined.
Polymorphisms of the CYP2C9 (cytochrome P450, family 2, subfamily C, polypeptide 9) gene (CYP2C9*2, CYP2C9*3) and the VKORC1 (vitamin K epoxide reductase complex, subunit 1) gene (-1639G>A) greatly ...impact the maintenance dose for the drug warfarin. Prescreening patients for their genotypes before prescribing the drug facilitates a faster individualized determination of the proper maintenance dose, minimizing the risk for adverse reaction and reoccurrence of thromboembolic episodes. With current methodologies, therapy can be delayed by several hours to 1 day if genotyping is to determine the loading dose. A simpler and more rapid genotyping method is required.
We developed a single-nucleotide polymorphism (SNP)-detection assay based on the SMart Amplification Process version 2 (SMAP 2) to analyze CYP2C9*2, CYP2C9*3, and VKORC1 -1639G>A polymorphisms. Blood from consenting participants was used directly in a closed-tube real-time assay without DNA purification to obtain results within 1 h after blood collection.
We analyzed 125 blood samples by both SMAP 2 and PCR-RFLP methods. The results showed perfect concordance.
The results validate the accuracy of the SMAP 2 for determination of SNPs critical to personalized warfarin therapy. SMAP 2 offers speed, simplicity of sample preparation, the convenience of isothermal amplification, and assay-design flexibility, which are significant advantages over conventional genotyping technologies. In this example and other clinical scenarios in which genetic testing is required for immediate and better-informed therapeutic decisions, SMAP 2-based diagnostics have key advantages.
The cyclic hexamer obtained by the condensation of furan and acetone can be used as a convenient starting material for the syntheses of calix6pyrrole 1 and of several “hybrid systems” containing both ...furan and pyrrole units. The solid‐state structure of 1 shows a water molecule held inside the cavity by six hydrogen‐bonding interactions, while that of the “alternating” calix3furan3pyrrole 2 contains no water molecules, despite the macrocycles having a similar structure.
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