A Study of the Effectiveness of Lidocaine to Treat Severe Pulmonary Vascular Constriction induced by Protamine

Background: Protamine neutralises heparin a?er separa?on from cardiopulmonary bypass. This study aimed to evaluate the effects of lidocaine on protamine induced pulmonary vascular constric?on in paediatric cardiothoracic surgery. Methods: This was a single-centre, prospec?ve, double-blind and randomised study conducted among eighty pediatric pa?ents with acyano?c congenital cardiac disease, scheduled for elec?ve on-pump cardiac surgery under general anaesthesia. In the study, the par?cipants were divided into four groups: Group NPHL- nonpulmonary hypertension with lidocaine precondi?oning, group NPHS- nonpulmonary hypertension with normal saline (as placebo), group PHL- pulmonary hypertension with lidocaine precondi?oning, and group PHS- pulmonary hypertension with normal saline (as placebo). Results: Pulmonary vasoconstric?on occurred in 11.25% of cases a?er protamine administra?on. Both the NPHS and PHS groups exhibited an increase in mean airway pressure (Paw), Respiratory index (RI), alveolar-arterial oxygen difference (A-aDO2), pulmonary artery pressure (PAP) and decreased dynamic pulmonary compliance (Cydn) and oxygen index (OI) a?er protamine administra?on. However, these changes were not observed in the NPHL and PHL groups with lidocaine precondi?oning. Plasma levels of TXB2 in the NPHS and PHS groups were higher than the NPHL and PHL groups, but 6-keto-PGF1 alpha levels were lower in the NPHS and PHS groups than in the NPHL and PHL groups. Conclusion: In congenital heart disease, repair without cardiopulmonary bypass is not possible in most cases. Prior to reversing heparin with protamine, precondi?oning lidocaine reverses protamine-induced pulmonary vasoconstric?on and improves lung func?on.

Influence of different levels of heparin and antithrombin Ⅲ on human coagulation factor Ⅸ activity / 中国输血杂志

【Objective】 To study the effect of different concentrations of heparin, ATⅢ or a mixture of heparin and antithrombin Ⅲ (ATⅢ) (1∶1)on the activity of human coagulation factor Ⅸ (FⅨ). 【Methods】 The heparin or heparin/ATⅢ with different concentrations were added into human coagulation Ⅸ products or human prothrombin complex (PCC) to prepare heparin or heparin/ATⅢ samples, containing 0, 0.1, 0.3, 0.5, 0.8, 1, 2 and 4 IU per unit. ATⅢ with different concentrations were added into FⅨ or PCC to prepare ATⅢ samples containing ATⅢ 0, 0.1, 0.5 and 1 IU per unit. The FⅨ activity of the samples prepared was tested by one-stage coagulation method. Then corresponding amount of protamine sulfate were added to neutralize heparin or heparin/ATⅢ to detect the FⅨ activity again. Their influence of heparin, ATⅢ and heparin/ATⅢ with different concentrations on the activity of FⅨ were analyzed. 【Results】 When the content of heparin or heparin/ATⅢ was 0, 0.1, 0.3 and 0.5 IU per unit of FⅨ, the detection results of FⅨ titer in samples were consistent. When the content of heparin or heparin/ATⅢ per unit of FⅨ was 0.8, 1, 2 and 4 IU, the detection results of FⅨ titer were all lower than those of samples without heparin. When the ATⅢ content was 0, 0.1, 0.5 and 1 IU, the FⅨ titer of the samples was consistent. 【Conclution】 When the content of heparin or heparin/ATⅢ in the product is less than or equal to 0.5 IU per IU of FⅨ, the step of protamine sulfate adding could be omitted as it has little effect on FⅨ activity. When >0.5 IU per IU of FⅨ, however, protamine sulfate adding, to neutralize heparin, is necessary before FⅨ activity testing.

Comparison of ART outcomes in men with altered mRNA protamine 1/protamine 2 ratio undergoing intracytoplasmic sperm injection with ejaculated and testicular spermatozoa / 亚洲男科学杂志(英文版)

Assisted reproductive technologies involving the use of spermatozoa and eggs for in vitro fertilization (IVF) have come as the solution for many infertile couples to become parents. However, in some cases, the use of ejaculated spermatozoa delivers poor IVF performance. Some studies have suggested the use of testicular spermatozoa in severe male infertility cases, but no guidelines regarding their utilization are currently available. In the present study, we found the mRNA protamine 1/protamine 2 (P1/P2) ratio to be a valuable biomarker of poor sperm function that could be used as a diagnostic key for the identification of cases that would benefit from the use of testicular spermatozoa. A total of 23 couples undergoing egg donation cycles with at least one previous cycle failure were studied. All couples underwent two consecutive intracytoplasmic sperm injection (ICSI) cycles with either ejaculated or testicular spermatozoa (TESA). The sperm mRNA P1/P2 ratio, fertilization rate, blastocyst rate, and pregnancy and live birth rate were compared. Results showed improved ICSI and clinical outcomes in cycles with testicular spermatozoa in men with altered mRNA P1/P2 ratios. TESA cycles presented significantly higher rates of fertilization (mean ± standard deviation: 76.1% ± 15.1% vs 65.5% ± 18.8%), blastocyst formation (55.0% ± 20.3% vs 30.8% ± 23.8%), and good morphological quality blastocyst (28.9% ± 22.9% vs 13.5% ± 17.9%) and also improvements on pregnancy (60.9% vs 0%) and healthy birth rates (56.5% vs 0%) than EJACULATE cycles. The results described here suggest that in patients with previous IVF/ICSI failures and aberrant mRNA protamine ratios, the use of testicular spermatozoa may be a good alternative to improve clinical outcomes.

Determination of Colistin Resistance by Simple Disk Diffusion Test Using Modified Mueller-Hinton Agar

BACKGROUND: Colistin has become a last-resort antibiotic for the management of multidrug-resistant gram-negative bacteria. The disk diffusion test is cheap and easy to perform but may be unreliable for colistin susceptibility testing due to poor diffusion of the large colistin molecule. An improved agar diffusion test would increase the reliability of colistin susceptibility testing. This study aimed to modify Muller-Hinton agar (MHA) to improve colistin diffusion in agar.METHODS: MHA was modified by reducing the agar concentration from 100% to 30% and supplementing with protamine. We tested 60 gram-negative clinical isolates of Pseudomonas aeruginosa (N=27) and Acinetobacter calcoaceticus-baumannii complex (N=33). Disk diffusion test results were interpreted based on minimum inhibitory concentrations determined by broth microdilution.RESULTS: The modified MHA yielded the best performance metrics, including 94.7% sensitivity, 100% specificity, and an area under the curve of 0.995 (95% confidence interval, 0.982–1.000), P<0.001, at a cut-off point of 13 mm.CONCLUSIONS: A reduction of the agar concentration from 100% to 30% and the addition of protamine improved colistin diffusion in agar and allowed routine colistin susceptibility testing in a clinical microbiology laboratory, but should be handled with caution.

Iodixanol supplementation during sperm cryopreservation improves protamine level and reduces reactive oxygen species of canine sperm

The objective of this study was to analyze the protective effects of iodixanol on dog spermatozoa during cryopreservation. The optimal concentration of iodixanol, 1.5%, was determined using fresh spermatozoa and was applied in the following experiments. The 1.5% iodixanol group showed significantly increased sperm motility from that in the control (p < 0.05). Lower mitochondrial reactive oxygen species (ROS) modulator (ROMO1) and pro-apoptotic gene (BAX) expressions, together with higher expressions of protamine-2 (PRM2), protamine-3 (PRM3), anti-apoptotic B-cell lymphoma-2 (BCL2), and sperm acrosome associated-3 (SPACA3) genes were detected in the iodixanol-treated group. In addition, decreased protamine deficiency and cryocapacitation were observed in the treatment group. Our results show that supplementation with 1.5% iodixanol is ideal for reducing production of ROS and preventing detrimental effects during the canine sperm cryopreservation process, effects manifested as increased motility and reduced cryocapacitation in frozen-thawed spermatozoa.

Study on Interaction of Protamine and Single-stranded DNA by Current Pulses Through Single Nanopore / 分析化学

Single nanopore current pulse method is a new, rapid and simple detection method, which is promising for single-molecule DNA sequencing and bio-sensing. Due to the short duration and the low current amplitude of the pulses caused by molecular translocation under normal conditions, pulse detection system with fast response and high sensitivity is required. In this work, based on a lab-established pulse detection system, the effect of protamine in the regulation of single-stranded (ssDNA) current pulses with α-hemolysin (α-HL) single nanopore interface was investigated. Experimental results showed that the pulses positive charged protamine and negative ssDNA probes were both well observed with the established system, and both the pulse amplitude and duration of ssDNA were increased as a result of interaction with protamine. This study provides a way to improve the resolving power of current pulses based on molecular interactions.

Clinical study of heparin protamine anticoagulant regimen optimization in patients undergoing cardiac surgery / 实用医学杂志

Objective To compare the effect of heparin protamine double titration with traditional meth-ods on postoperative bleeding in patients underwent cardiopulmonary bypass(CPB). Methods Ninety adult patients scheduled to elective CPB by open heart surgery were randomly divided into three groups. In the Conven-tional Group(group A,n=30),375 U/kg heparin was administered.The amount of protamine was in proportion to 125 U:1.2 mg base on the initial heparin dose.In the Heparin Titration Group(group B,n=30),heparin was given according to the effect of heparin titration. In the Heparin and Protamine Double Titration Group(group C, n = 30),heparin titration combined with protamine titration were administered. The general data of the patients were recorded. The blood transfusion volume during and after operation,volume of thoracic drainage within 24 hours after operation,operative complications were compared among the 3 groups. Results Group B and group C heparin dosage were less than the group A(P<0.05).The protamine dosage in group C was less than that in group A and group B(P < 0.05). The thoracic drainage volume within 24 hours after operation was higher in the group A than group B and group C(P < 0.05). Conclusion Compared the traditional anticoagulation with heparin and protamine double titration,the latter individualized anticoagulation can reduce the amount of bleeding after CPB.

Cost-minimization Analysis of Four Kinds of Premixed Insulin in the Therapy for Newly Diagnosed Type 2 Diabetes / 中国药师

Objective:To evaluate the short-term economic effects of four kinds of premixed insulin in newly diagnosed type 2 dia-betes mellitus. Methods:A total of 120 newly diagnosed patients with type 2 diabetes mellitus were divided into four groups according to the kind of premixed insulin, group A was treated with insulin aspart 30 injection, group B was treated with insulin lispro 25 injec-tion, group C was treated with isophane protamine biosynthetic human insulin injection and group D was treated with protamine zinc re-combinant human insulin injection. The course of treatment was three months. The therapy efficacy was assessed by the remission rate in three months. The short-term economic effect was evaluated by the cost-minimization analysis method. Results:The remission rate of group A, B, C and D respectively was 48. 39%, 48. 28%, 51. 61% and 51. 72% without significant difference (P>0. 05). The average cost per person of the four groups was 1195. 52, 1202. 41, 1220. 69 and 1258. 84 yuan, and the average medicine cost per person was 750. 52, 689. 41, 754. 69 and 764. 34 yuan, respectively. There was no significant difference in cost among the four groups (P >0. 05). Conclusion:All the four kinds of premixed insulin can be used for the starting treatment with the similar total cost, and in relative terms, aspart 30 injection and insulin lispro 25 injection are better for the initial treatment of diabetes.

Qualities Study of Protamine Sulfate Raw Material from Different Species of Fish / 中国药学杂志

OBJECTIVE: To study the qualities of protamine sulfate raw material from different species of fish. METHODS: The identification and determination of protamine sulfate raw material from different fish species were conducted by HPLC, and the composition of amino acids was tested too. Heavy metals was tested by inductively coupled plasma mass spectrometry. The potency of protamine sulfate was measured by biological assay method. RESULTS: At present, the sources of protamine sulfate on the market were mainly salmon and herring. CONCLUSION: This HPLC method can be applied to effectively identify and determine protamine sulfate raw material in order to improve its quality control.

Evaluation of sperm protamine deficiency and apoptosis in infertile men with idiopathic teratozoospermia / 대한생식의학회지

OBJECTIVE: Sperm morphology plays an important role in infertility, especially in cases of defects in the heads of spermatozoa. Tapered-head or elongated-head spermatozoa are examples of morphological abnormalities. The aim of this study was to compare the semen parameters, levels of protamine deficiency, and frequency of apoptosis between patients with normozoospermia and those with teratozoospermia with tapered-head spermatozoa. METHODS: Fifty-two semen samples (27 patients with tapered-head sperm and 25 fertile men) were collected and semen analysis was performed according to the World Health Organization criteria for each sample. Protamine deficiency and the percentage of apoptotic spermatozoa were evaluated using chromomycin A3 (CMA3) staining and terminal deoxynucleotidyl transferase dUTP nick-end labelling (TUNEL) assays, respectively. RESULTS: Sperm concentration, motility, and normal morphology in the tapered-head spermatozoa (cases) were significantly lower than in the normozoospermic samples (controls). CMA3-reactive spermatozoa (CMA3+) in the case group were more common than in the controls. Apoptotic spermatozoa (TUNEL-positive) were significantly more common in the cases than in the controls. CONCLUSION: This analysis showed that tapered-head spermatozoa contained abnormal chromatin packaging and exhibited a high rate of apoptosis, which can be considered to be an important reason for the impaired fertility potential in teratozoospermic patients with tapered-head spermatozoa.

Effect of Protamine Coated PLA Nanocapsule on the Immune Function of Murine BMDCs In Vitro / 华中科技大学学报(医学版)

Objective To investigate the effect of protamine(PS)coated polylactic acid(PLA)nanocapsule on the immune response of murine bone marrow-derived dendritic cells(BMDCs).Methods PLA nanocapsules were prepared by W1/O/W2 method.Protamine was used to modify the surface of PLA nanocapsule that encapsulated ovalbumin(OVA),a model anti-gen.The prepared OVA/PLA/PS nanocapsules were used to stimulate BMDCs.Flow cytometry was used to analyze the ex-pression of surface molecules CD80,CD83,CD86,MHCⅠ,and MHC Ⅱ,and the uptake of nanocapsule.The levels of IL-12 p70 secreted by BMDCs were detected by ELISA.Results OVA/PLA/PS nanocapsules could significantly up-regulate the expres-sion of CD80,CD83,MHCⅠ,and MHC Ⅱ in BMDCs,and increase the secretion of IL-12 p70 by BMDCs.Furthermore,OVA/PLA/PS nanocapsules could enhance the uptake efficiency of OVA by BMDCs.Conclusion OVA/PLA/PS nanocapsule can en-hance the immune response of BMDCs,and may become a good drug delivery carrier.

Research and Demonstration of DUE Mode in the Clinical Utilization Evaluation of Insulin / 中国药师

Objective:To establish the drug utilization evaluation(DUE)criteria for isophane protamine biosyntheti(30R)to provide reference for the rational drug use. Methods:Referred to the domestic and foreign literatures and the authorized guidelines for diabetes treatment,and combined with the Delphi expert consultation method,the drug utilization evaluation criteria was established,and after the evaluation,intervention and revaluation,the final criteria were determined. Results:The drug utilization evaluation criteria for isophane protamine biosyntheti(30R)contained three parts,including medication indications,medication process and medication results,which showed the clinical feasibility. Conclusion:The established DUE criteria for isophane protamine biosyntheti(30R)can not only be applied in the drug utilization evaluation,but also provide reference for the DUE in medical institutions.

Nose-to-brain delivery of macromolecules mediated by cell-penetrating peptides

Brain delivery of macromolecular therapeutics (e.g., proteins) remains an unsolved problem because of the formidable blood-brain barrier (BBB). Although a direct pathway of nose-to-brain transfer provides an answer to circumventing the BBB and has already been intensively investigated for brain delivery of small drugs, new challenges arise for intranasal delivery of proteins because of their larger size and hydrophilicity. In order to overcome the barriers and take advantage of available pathways (e.g., epithelial tight junctions, uptake by olfactory neurons, transport into brain tissues, and intra-brain diffusion), a low molecular weight protamine (LMWP) cell-penetrating peptide was utilized to facilitate nose-to-brain transport. Cell-penetrating peptides (CPP) have been widely used to mediate macromolecular delivery through many kinds of biobarriers. Our results show that conjugates of LMWP-proteins are able to effectively penetrate into the brain after intranasal administration. The CPP-based intranasal method highlights a promising solution for protein therapy of brain diseases.

Comparison and analysis of two potency assay methods of protamine sulfate / 中国生化药物杂志

Objective To analyze and compare two potency assay methods of protamine sulfate.Methods Heparin titration method was according to foreign pharmacopoeia,while biological assay method was according to China pharmacopoeia(2015).Results The absorbance at the end-point was stable at 0.10 mg/mL and 0.15 mg/mL test solutions.The relative standard deviations(RSDs)of potency results were all less than 5%.Three test solutions and volumes of the titrant had a good linear relationship(R2 =1.000).The results of titration method were significantly related to those of biological assay(P=0.013),with similar RSD(P>0.05).However,the potency of titration method were significantly lower than those of biological assay (P=0.045).Conclusion Heparin titration method is good-using,convenient,and need to be applied widely at home.

Effect of insulin injection and protamine biosynthetic human insulin injection on basal insulin level in pregnant women with diabetes mellitus / 中国生化药物杂志

Objective To explore the effect of insulin injection and protamine biosynthetic human insulin injection on basal insulin level in pregnant women with diabetes mellitus.Methods Retrospective analysis of 89 cases of pregnant women with diabetes mellitus from January 2013 to May 2016 in department of obstetrics and gynecology,tianjin red bridge hospital,the patients were divided into group A (n=38 cases) and group B (n=51 cases),the group A treatment with insulin injection,the group B treatment with protamine biosynthetic human insulin injection,compare the two groups of patients before and after treatment of three meals a day rate of blood glucose compliance, treatment compliance and satisfaction.Results Before treatment, there was no significant difference in the blood glucose compliance rate between the two groups before and after treatment;After treatment,the blood glucose compliance rate of two groups was significantly higher than before treatment ( P<0.05 ) , there was no significant difference in the compliance rate of fasting blood glucose between two groups,the compliance rate of blood glucose before dinner in group A was 81.58%,significantly higher than that in group B 60.78%(P<0.05).Conclusion Both insulin injection and protamine biosynthetic human insulin injection can maintain the basic insulin levels of pregnant women with diabetes mellitus,insulin injection can better control the blood glucose levels before dinner,with higher compliance and satisfaction.The compliance rate and satisfaction rate of pregnant women in group A were 97.37% and 97.37%,which were significantly higher than those in group B 82.35% and 80.39%(P<0.05).

Association study of PRM2 gene polymorphism with male infertility in the 386 Chinese Han population / 重庆医学

Objective To determine the association of one single nucleotide polymorphism loci G 398C in PRM2 with male in‐fertility in Chinese Han population .Methods A total of 386 infertile men were recruited as the observation group and 255 fertile men were recruited as the control group .Routine semen analysis as well as sperm functional parameters such as DNA integrity and nucleoprotein maturity rates were analyzed .Direct sequencing of G398C in PRM2 gene of infertile and fertile men was also conduc‐ted to evaluate the association of G398C SNP loci with male infertility .Results Statistical analysis showed that the frequencies of CC genotype of PRM2 G398C was significantly different between the infertile (11 .92% ) and fertile men (6 .67% ) and it was asso‐ciated with increased risk of male infertility (OR= 2 .002 ,95% CI= 1 .097 -3 .653 ,P< 0 .05) .Moreover ,it was discovered that sperm DNA integrity as well as nucleoprotein maturity rate of CC genotype were dramatically decreased compared with other geno ‐types (P<0 .05) ,which would probably lead to infertility .Conclusion Our results gave the first evidence that PRM 2 G398C poly‐morphism was associated with male infertility in Chinese Han population .

Preparation and Quality Control of Coenzyme Q10 Long-circulating Liposomes and Lyophilized Prepara-tion / 中国药房

OBJECTIVE:To prepare Coenzyme Q10 long-circulating liposomes,establish the determination method of content and entrapment efficiency,and prepare it into lyophilized preparation to improve its stability. METHODS:Coenzyme Q10 long-cir-culating liposomes were prepared by film dispersion method. Particle size and Zeta potential of liposomes were determined,and HPLC assay was used to determine the content of coenzyme Q10. Free drugs and liposomes were separated using protamine aggre-gation method,and the encapsulation efficiency was calculated. Lyophilized preparation was prepared by coenzyme Q10 long-circu-lating liposomes,and the changes of content and encapsulation efficiency of drugs were determined 0,30 and 90 days after lyophi-lization. RESULTS:The liposomes were homogeneous in size with mean diameter of(166.0±5.3)nm and Zeta potential of(-22.2± 1.4)mV. Average content(the percentage of content accounted for labeled amount)and entrapment efficiency of 3 batches of sam-ple were 98.2%(RSD=2.8%) and 93.2%(RSD=4.6%),respectively. Compared with 0 d after lyophilization,coenzyme Q10 long-circulating liposomes had no obvious change in the content and encapsulation efficiency 90 d after lyophilization. CONCLU-SIONS:Coenzyme Q10 long-circulating liposomes with high quality and entrapment efficiency and lyophilized preparation being stored stably for 90 d have been prepared successfully.

Efficient isolation of sperm with high DNA integrity and stable chromatin packaging by a combination of density-gradient centrifugation and magnetic-activated cell sorting / 대한생식의학회지

OBJECTIVE: This study was carried out to investigate the correlations of the sperm DNA fragmentation index (DFI) with semen parameters and apoptosis, and to investigate the effects of density-gradient centrifugation (DGC) and magnetic-activated cell sorting (MACS) on reducing the proportion of sperm with DNA fragmentation and protamine deficiency. METHODS: Semen analysis and a sperm DNA fragmentation assay were performed to assess the correlations between semen parameters and the DFI in 458 semen samples. Sperm with progressive motility or non-apoptosis were isolated by DGC or MACS, respectively, in 29 normozoospermic semen samples. The effects of DGC or MACS alone and of DGC and MACS combined on reducing the amount of sperm in the sample with DNA fragmentation and protamine deficiency were investigated. RESULTS: The sperm DFI showed a significant correlation (r=–0.347, p<0.001) with sperm motility and morphology (r=–0.114, p<0.05) but not with other semen parameters. The DFI (11.5%±2.0%) of semen samples was significantly reduced by DGC (8.1%±4.1%) or MACS alone (7.4%±3.9%) (p<0.05). The DFI was significantly further reduced by a combination of DGC and MACS (4.1%±1.3%, p<0.05). Moreover, the combination of DGC and MACS (1.6%±1.1%, p<0.05) significantly reduced the protamine deficiency rate of semen samples compared to DGC (4.4%±3.2%) or MACS alone (3.4%±2.2%). CONCLUSION: The combination of DGC and MACS may be an effective method to isolate high-quality sperm with progressive motility, non-apoptosis, high DNA integrity, and low protamine deficiency in clinical use.

Impaired protamination and sperm DNA damage in a Nellore bull with high percentages of morphological sperm defects in comparison to normospermic bulls / Deficiências de protaminação e danos de DNA no sêmen de um touro Nelore com elevada porcentagem de patologias espermáticas em comparação a touros normospérmicos

The routine semen evaluation assessing sperm concentration, motility and morphology, does not identify subtle defects in sperm chromatin architecture. Bulls appear to have stable chromatin, with low levels of DNA fragmentation. However, the nature of fragmentation and its impact on fertility remain unclear and there are no detailed reports characterizing the DNA organization and damage in this species. The intensive genetic selection, the use of artificial insemination and in vitro embryo production associated to the cryopreservation process can contribute to the chromatin damage and highlights the importance of sperm DNA integrity for the success of these technologies. Frozen-thawed semen samples from three ejaculates from a Nellore bull showed high levels of morphological sperm abnormalities (55.8±5.1%), and were selected for complementary tests. Damage of acrosomal (76.9±8.9%) and plasma membranes (75.7±9.3%) as well as sperm DNA strand breaks (13.8±9.5%) and protamination deficiency (3.7±0.6%) were significantly higher compared to the values measured in the semen of five Nellore bulls with normospermia (24.3±3.3%; 24.5±6.1%; 0.6±0.5%; 0.4±0.6% for acrosome, plasma membrane, DNA breaks and protamine deficiency, respectively) (P<0.05). Motility and percentage of spermatozoa with low mitochondrial potential showed no differences between groups. This study shows how routine semen analyses (in this case morphology) may point to the length and complexity of sperm cell damage emphasizing the importance of sperm function testing.(AU)

O exame de rotina de sêmen, o qual avalia a concentração de espermatozoides, a motilidade e a morfologia, pode não identificar defeitos sutis na arquitetura da cromatina de espermatozoides. Os touros parecem ter cromatina estável com baixos níveis de fragmentação do DNA. No entanto, a natureza da fragmentação e o seu impacto sobre a fertilidade ainda não estão claros e não há relatos que caracterizam a organização do DNA e os danos nessa espécie com mais detalhes. A seleção genética intensiva e o uso da inseminação artificial e da produção in vitro de embriões, além do processo de criopreservação, podem contribuir para o dano da cromatina, e sabe-se a importância da integridade do DNA espermático para o sucesso dessas tecnologias. Amostras de sêmen de três ejaculados de um touro Nelore com altos níveis de alterações morfológicas (55,8±5,1%) foram selecionadas para realização de exames complementares. Os danos de acrossoma (76,9±8,9%) e das membranas plasmáticas (75,7±9,3%), bem como quebras de fita de DNA de espermatozoides (13,8±9,5) e deficiência de protamina (3,7±0,6) foram significativamente maiores em comparação aos valores avaliados no sêmen de cinco touros Nelore com normospermia (24,3±3,3%; 24,5±6,1%; 0,6±0,5%; 0,4±0,6% para acrossoma, membrana plasmática, quebras de DNA e deficiência de protamina, respectivamente) (p<0,05). Motilidade e porcentagem de espermatozoides com baixo potencial mitocondrial não diferiram estatisticamente. Essas avaliações mostram que análises de sêmen de rotina (neste caso, morfologia) podem apontar para a extensão e a complexidade dos danos na célula espermática, o que indica que a deficiência de protamina e os danos no DNA podem ocorrer simultaneamente a defeitos morfológicos. Tal ocorrência enfatiza a importância das análises de sêmen clássicas e dos testes complementares.(AU)

Protective Effects of CYP2E1 Inhibitors on Metabolic Syndrome-induced Liver Injury in Guinea Pigs.

The present work reports the effects of CYP2E1-inhibitors (quercetin, 4- methylpyrazole and disulfiram) on the indices characterizing state of the liver in guinea pigs with metabolic syndrome (MS) induced by protamine sulfate repeated administrations. The investigation of quercetin, 4-methylpyrazole and disulfiram effects on hepatic cytochrome P450 2E1 (CYP2E1) protein and activity changes was conducted. Simultaneously, the content of reactive oxygen species (ROS) and markers of liver damage were determined in experimental animals’ blood. The link between increased hepatic expression of CYP2E1, prominent ROS generation and liver damage in animals with MS has been discovered. It has been demonstrated that CYP2E1 protein content and activity in guinea pigs with MS rose almost 3 times compared to intact animals. These events were accompanied by increase in ROS generation and metabolism and liver disturbances symptoms: increase in serum glucose and cholesterol contents (2 and 2.6 times respectively), alanine aminotransferase (2.8 times), aspartate aminotransferase (6.4 times), and alkaline phosphatase (1.8 times) elevations. Our investigation suggests that administration of quercetin, 4-methylpyrazole, and disulfiram in guinea pigs with MS caused decrease in this isoenzyme protein expression (2.5, 1.7 and 2.4 respectively) as well as its enzymatic activity in liver (6.6, 1.1, and 1.7 respectively). The content of blood ROS was partially restored or normalized by all three CYP2E1 inhibitors. In turn, suppression of CYP2E1 activity and ROS generation led to decrease in hepatic MS manifestation. It is apparent from the present observation that quercetin has the highest efficiency among the investigated substances. Further studies on various quercetin doses and administration regimens could provide relevant information for the development of MSrelated nonalcoholic fatty liver disease treatment.