Tobacco smoking and its impact on the expression level of sperm nuclear protein genes: H2BFWT, TNP1, TNP2, PRM1 and PRM2.

The aim of this current study was to investigate the influence of tobacco smoke on sperm quality determined by standard parameters, on sperm DNA maturity tested by chromomycin A3 (CMA3) staining, on sperm DNA fragmentation tested by TUNEL assay and on the transcript level of sperm nuclear proteins H2BFWT, PRM1, PRM2, TNP1 and TNP2 genes quantified by RT-PCR. One hundred forty-one (141) sperm samples (43 nonsmokers (G.1) and 98 heavy smokers (G.2)) of couples undergoing ICSI were enrolled in this study. In G2, a significant decrease in standard semen parameters in comparison with nonsmokers was shown (p < .01). In contrast, protamine deficiency (CMA3 positivity) and sperm DNA fragmentation (sDF) were significantly higher in G2 than in G1 (p < .01). Furthermore, the studied genes were differentially expressed (p < .01), down-regulated in the spermatozoa of G.2 compared to that of G.1 (fold change <0.5) and were significantly correlated between each other (p < .01). Moreover, in comparison with G1, the protamine mRNA ratio in G2 was significantly higher (p < .01). It can therefore be concluded that smoking alters mRNA expression levels of H2BFWT, TNP1, TNP2, PRM1 and PRM2 genes and the protamine mRNA ratio and consequently alters normal sperm function.

An additional marker for sperm DNA quality evaluation in spermatozoa of male partners of couples undergoing assisted reproduction technique (IVF/ICSI): Protamine ratio.

The aim of this study was to evaluate the relationship between the protamine ratio (P1/P2), DNA fragmentation of spermatozoa and protamine deficiency. Patients were grouped into fertile (G1; n = 151) and sub-fertile (G2; n = 121). DNA fragmentation in spermatozoa was analysed by a TUNEL assay (terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick-end labelling), and the protamination was determined by CMA3 staining, while Western blot was used to measure protamine P1 and P2. While sperm DNA fragmentation (SDF) and protamine ratio were significantly elevated in G2 compared with G1 (12.31 ± 7.01% vs. 17.5 ± 9.5%; p = .001) and (0.91 ± 0.43 vs. 0.75 ± 0.42; p = .003); respectively, the CMA3 positive showed no difference at all between G1 and G2. In G1, the CMA3 positive correlated negatively with the P1/P2 ratio and SDF (r = -.586, r = -.297; p = .001 respectively). In contrast, the protamine ratio correlated positively with SDF (r = .356; p = .001). In G2, no correlation was observed between CMA3 positive, SDF and the P1/P2 ratio but the P1/P2 ratio showed a positive correlation with SDF (r = .479; p = .001). In conclusion, the spermatozoa DNA deterioration was closely associated with abnormal protamination but showed an association with the protamine ratio, more than with CMA3 positive. Therefore, for the evaluation of DNA damage in spermatozoa, the P1/P2 ratio might act as an additional biomarker.

A gap analysis of SDG 3 and MDG 4/5mortality health targets in the six Arabic countries of North Africa: Egypt, Libya, Tunisia, Algeria, Morocco, and Mauritania.

BACKGROUND: The United Nations Assembly adopted the Sustainable Development Goals to succeed the Millennium Development Goals in September 2015. From a European perspective, the development of health in the countries of North Africa are of special interest as a critical factor of overall social development in Europe's Mediterranean partners. In this paper, we address the mortality related SDG-3 targets, the likelihood to achieve them until 2030 and analyze how they are defined. METHODS: We projected mortality trends from 2000-2015 to 2030, based on mortality estimates by inter-agency groups and the WHO in mother and child health, non-communicable diseases, and road traffic mortality. The gap analysis compares the time remaining until 2030 to the time needed to complete the target assuming a linear trend of the respective indicator. A delay of not more than 3.75 years is considered likely to achieve the target. RESULTS: The SDG-3 targets of a Maternal Mortality Ratio below 70 per 100 000 live births and an U5MR below 25 per 1 000 live births have been achieved by Egypt, Libya, and Tunisia. Libya and Tunisia have also achieved the target for Newborn Mortality with Egypt close to achieving it as well. Algeria and Morocco are generally on track for most of the indicators, including deaths from non-communicable diseases and suicide rates; however, all of the countries are lagging when it comes to deadly Road Traffic Injuries for 2030. Mauritania is the only North African country which is not likely to reach the 2030 targets for any of the mortality indicators. CONCLUSIONS: Although mortality statistics may be incomplete there is an impressive gradient from East to West showing Mauritania and deadly road traffic injuries as the most problematic areas. Given the large differences between countries baselines, we consider it preferable to set realistic targets to be achieved until 2030.

Effect of acupotomy versus electroacupuncture on ethology and morphology in a rabbit model of knee osteoarthritis.

OBJECTIVE: To evaluate the treatment effect of acupotomy (Apo) in a rabbit model of knee osteoarthritis (KOA), compare the results of Apo versus electroacupuncture (E-Apu) on ethology, morphology, and structure of the articular cartilage surface in a rabbit model of KOA, and analyze the differences in the treatment effects of Apo versus E-Apu. METHODS: Twenty-eight male New Zealand white rabbits were randomized into four groups: normal control, blank model, Apo, and E-Apu (n = 7 in each group). Except for the normal control group, the left hindlegs of all rabbits were fixed in an extended position for 5 weeks to establish the KOA model. The passive range of motion (PROM) and Lequesne index were measured before and after the establishment of the KOA model to assess the ethology in all groups. Safranin O-fast green staining and the Mankin score were used to assess the morphological cartilaginous changes to compare the effect of Apo versus E-Apu on the degeneration of articular cartilage, and to identify which therapy was superior in treating KOA. RESULTS: Compared with before the establishment of the KOA model, the Lequesne index of the KOA model rabbits was significantly increased (P < 0.01), while the PROM was significantly decreased (P < 0.01). The articular cartilaginous tissue in the three model groups exhibited pathological variations in the form of laminar derangement of cartilage cells, and so the Mankin score was significantly increased compared with the control group (P < 0.01). At 1 week after the final treatment session, compared with the blank model group, both the Apo and E-Apu groups showed a significant decrease in the Lequesne index (P < 0.01), and attenuation in the degree of morphologic pathological changes (P < 0.05); The Apo improved the PROM significantly compared with the blank model group (P < 0.05), while the E-Apu had no effect (P > 0.05). Furthermore, compared with the E-Apu group, the Apo group had a significantly lower Lequesne index (P < 0.05), and a significantly greater PROM (P < 0.05). CONCLUSION: In a rabbit model of KOA, both Apo and E-Apu reduce disorders of ethology and morphology, and improve the condition of the articular cartilage. The results suggest that Apo is more effective than E-Apu in improving the PROM and alleviating symptoms resulting from cartilage damage in a rabbit model of KOA.

The relationship between preimplantation morphokinetics of human embryos and sex chromosome pattern.

The aim of this study was to determine the relationship between the gender of human embryos and chronological changes during the two pronuclear (2PN) embryonic stage and blastocyst formation on day five after injection using time-lapse imaging and preimplantation genetic testing. A total of 120 couples who underwent intracytoplasmic sperm injection with preimplantation genetic testing were included in the study. Only normal embryos (n = 416) were enrolled in this study. Time-lapse imaging data of male (n = 227) and female (n = 189) embryo progression was carried out to estimate the times of initial appearance of the 2PN stage, fading times and day five blastulation rates. The results revealed that the 2PN stage (fertilisation) was reached significantly earlier in female embryos (9.09 ± 1.31 h) than in male embryos (9.52 ± 1.48 h, p-value = 0.0044). Conversely, the fading time of 2PN was significantly faster in male embryos (22.13 ± 2.02) than in female embryos (24.16 ± 2.61, p-value < 0.001). The blastulation rate was significantly higher in female than male embryos (68.25% vs. 57.71%; p-value = 0.025).

Relationships between bacteriospermia, DNA integrity, nuclear protamine alteration, sperm quality and ICSI outcome.

The Aim of this study was to evaluate the effects of bacteriospermia on human sperm parameters, nuclear protamines, DNA integrity and ICSI outcome in patients enrolled for ICSI treatment. 84 unselected couples consulting in infertility and obstetrics clinic and enrolled for ICSI treatment were included in this study. The semen specimens were screened bacteriologically; semen and sperm parameters were also evaluated according to WHO guidelines. DNA integrity, protamines concentration and protamine deficiency were estimated by TUNEL assay, AU-PAGE and Chromomycin (CMA3) respectively. The results of this study revealed that 34.52% of studied semen samples were infected with bacteria. The isolated bacteria were identified as Staphylococcus aureus, Staph. epidermidis, Staph. haemolyticus, Escherichia coli, Enterococcus faecalis and Streptococcus agalactiae. Bacteriospermia had a significant (p < .010) negative effect on sperm parameters; concentration, motility, progressive motility and chromatin condensation. Moreover, high DNA fragmentation with low P1 and P2 concentrations were noticed in infected patients in comparison to non-infected patients but non-significant. Also, the fertilization rate decreased significantly (p < .05) with infected patients. IN CONCLUSION: bacteriospermia has significant negative effect on sperm quality and fertilization rate in patients who underwent ICSI treatment.