Oxidative stress, hepcidin and nesfatin-I status in childhood iron and vitamin B12 deficiency anemias.

BACKGROUND: Anemia is a disease that is long and often repetitive and can result in a great burden to the national economy. The most frequent nutritional deficiency anemias in children are related with iron and vitamin B12 deficiencies. OBJECTIVES: The aim of this study was to determine the oxidative stress, hepcidin, and nesfatin-I levels in childhood iron and vitamin B12 deficiency anemias. MATERIAL AND METHODS: The study had 3 groups of 15 children, iron anemia deficiency group, vitamin B12 deficiency group and a control group. RESULTS: The TBARS and nesfatin-I levels were significantly higher in the iron and vitamin B12 deficiency groups and the total antioxidant levels were significantly lower when compared to the control group. In contrast, the plasma hepcidin levels were significantly lower in the iron deficiency group (p < 0.01) when compared to the control group; however, no significant differences were observed in the vitamin B12 deficiency group. Plasma homocysteine levels were significantly higher in the vitamin B12 deficiency group when compared to the control group (p < 0.001), but no differences were determined between the iron deficiency and control groups. CONCLUSIONS: Our results showed that there are high levels of oxidative stress in childhood iron and vitamin B12 deficiency anemias, and we propose that plasma hepcidin and homocycteine levels may be useful in the differential diagnosis of childhood nutritional deficiency anemias. Nesfatin-1 hormone levels were identified for the first time in childhood iron deficiency and vitamin B12 deficiency anemias within this study and this hormone may also be useful in the differential diagnosis of anemias.

Alteration of the fecal microbiota and serum metabolite profiles in dogs with idiopathic inflammatory bowel disease.

Idiopathic inflammatory bowel disease (IBD) is a common cause of chronic gastrointestinal (GI) disease in dogs. The combination of an underlying host genetic susceptibility, an intestinal dysbiosis, and dietary/environmental factors are suspected as main contributing factors in the pathogenesis of canine IBD. However, actual mechanisms of the host-microbe interactions remain elusive. The aim of this study was to compare the fecal microbiota and serum metabolite profiles between healthy dogs (n = 10) and dogs with IBD before and after 3 weeks of medical therapy (n = 12). Fecal microbiota and metabolite profiles were characterized by 454-pyrosequencing of 16 S rRNA genes and by an untargeted metabolomics approach, respectively. Significantly lower bacterial diversity and distinct microbial communities were observed in dogs with IBD compared to the healthy control dogs. While Gammaproteobacteria were overrepresented, Erysipelotrichia, Clostridia, and Bacteroidia were underrepresented in dogs with IBD. The functional gene content was predicted from the 16 S rRNA gene data using PICRUSt, and revealed overrepresented bacterial secretion system and transcription factors, and underrepresented amino acid metabolism in dogs with IBD. The serum metabolites 3-hydroxybutyrate, hexuronic acid, ribose, and gluconic acid lactone were significantly more abundant in dogs with IBD. Although a clinical improvement was observed after medical therapy in all dogs with IBD, this was not accompanied by significant changes in the fecal microbiota or in serum metabolite profiles. These results suggest the presence of oxidative stress and a functional alteration of the GI microbiota in dogs with IBD, which persisted even in the face of a clinical response to medical therapy.

Comparing ethylene glycol with glycerol and with or without dithiothreitol and sucrose for cryopreservation of bull semen in egg-yolk containing extenders.

There are few studies performed for investigating the roles of different ratio and cryoprotectants with dithiothreitol or sucrose on sperm motility characteristics and antioxidant capacities of post-thawed bull spermatozoa. The objectives of this study were to compare glycerol (G) and ethylene glycol (EG) at different concentrations as cryoprotectants and dithiothreitol (D) or sucrose (S) (with/without) as antioxidants in Tris extender for cryopreservation of bull semen. Twenty-four ejaculates obtained from three bulls were included in the study. Each ejaculate was split into four equal aliquots and diluted using both of the Tris extenders with glycerol (5% or 7%) or ethylene glycol (3% or 5%). After that, each extenders were split into three equal aliquots and diluted using both of the dithiothreitol 5mM or sucrose 25 mM, and control (without additives) was cooled to 4 °C and frozen in 0.25-ml French straws. when compared to control, different doses cryoprotectants and antioxidants addition no significantly increased the percentages of post-thaw sperm progressive and motitilities, acrosome abnormality and plasma membrane integrity (P>0.05). However, EG3+S yielded the greatest percentages of the total abnormality (P<0.05). As regard to antioxidant activities G7 and EG5 led to lowest MDA activity with or without D or S but, these results were not supported to the GPx activity (P<0.01). The sperm motion characteristics such as VAP, VCL, ALH and BCF gave significantly different results (P<0.05). When compared the DNA integrity, different doses cryoprotectants without antioxidants addition significantly increased the percentages of the tail intensity and tail moment (P<0.05). There were no significant differences observed in non-return rates among all treatment groups (P>0.05).

Can linoleic acid improve the quality of frozen thawed bull sperm?

BACKGROUND: Cryopreservation is known to have a detrimental effect on the motility, viability and membrane integrity of sperm cells. OBJECTIVE: The aim of this study was to investigate the effect of various amount of linoleic acid supplementation to the Tris extender, on bull sperm parameters, DNA integrity and oxidative stress after freeze-thawing. METHODS: Ejaculates were split into five aliquots and extended to a final concentration of 18x10(6) spermatozoa per ml with the base extender containing different doses of linoleic acid 0.125 ml, (L125); 0.250 ml (L250); 0.5 ml (L500), 1 ml (L1000) and no additive (control; L0). The extended samples were equilibrated slowly to 4 degree C for 4 h and then froze using a digital freezing machine. Frozen straws were thawed individually in water bath at 37 degree C for 30 s to analyse progressive motility and sperm motion characteristics as well as membrane integrity. Biochemical assays were performed in a spectrophotometer using commercial kits. DNA damage was evaluated by Comet Assay. RESULT: The addition of various linoleic acid did not improve the sperm subjective, CASA and progressive motilities, sperm motility characteristics and DNA integrity (P>0.05). L500 exhibited the greatest values for membrane integrity than that of the other groups (P<0.001). All supplementation groups led to lower percentages of tail abnormalities in comparison to the control (P<0.001). L500 and L1000 significantly decreased total abnormalities. In conclusion, our findings showed that L500 linoleic acid supplementation in semen extender was of great beneficial effect on frozen-thawed bull semen in terms of morphology and plasma membrane integrity.

Effects of various cryoprotectants on bull sperm quality, DNA integrity and oxidative stress parameters.

The objectives of this study was to compare the effects of type and concentration of cryoprotectants glycerol (G), ethylene glycol (EG) and dimethyl sulfoxide (DMSO) on the plasma membrane and DNA integrity as well as antioxidant activity of cryopreserved Eastern Anatolian red bull sperm. Ejaculates were collected from the three bulls using an artificial vagina twice a week. The ejaculates were pooled to increase the semen volume for replication and to eliminate variability among the evaluated samples. The pooled ejaculates were also split into seven equal experimental groups and diluted with the modified base extender to a final spermatozoa concentration of 15×10(6)/ml. The extended samples were cooled slowly to 4°C and equilibrated for 4h. They were then loaded into 0.25ml French straws and frozen using a digital freezing machine at 3 programmed rates: -3°C/min from +4°C to -10°C, -40°C/min from -10°C to -100°C, and -20°C/min from -100°C to -140°C. Thereafter, the straws were plunged into liquid nitrogen at -196°C. Frozen straws were thawed individually at 37°C for 30s in a water bath to analyse progressive motility and sperm motion characteristics as well as membrane integrity using hypo-osmotic swelling test. Biochemical assays were performed in a spectrophotometer using commercial kits. DNA damage was evaluated by Comet Assay using Image Analysis System. 6% G exhibited the greatest percentages of CASA (43.7±2.92%) and progressive (26.4±2.64%) motilities when compared to the other groups (P<0.001). 6% G and 6% EG showed the greatest values of preserved membrane integrity (P<0.001). 6% DMSO and 3% EG + 3% DMSO resulted in greater chromatin damage than the other groups (P<0.001). The antioxidant activities of GPx, GSH, and CAT as well as the total antioxidant activity were affected by the type of cryoprotectant; notably, 2% G+2% EG+2% DMSO yielded the lowest activities when compared to the other groups (P<0.001). In conclusion, no advantages were found in using EG or DMSO to replace G in bull sperm cryopreservation. Freezing with cryoprotectant 6% G yielded the best post-thaw sperm characteristics for Eastern Anatolian Red bull spermatozoa.

The effect of raffinose and methionine on frozen/thawed Angora buck (Capra hircus ancryrensis) semen quality, lipid peroxidation and antioxidant enzyme activities.

The aim of the present study was to determine the effects of different doses of raffinose and methionine on post-thawed semen quality, lipid peroxidation and antioxidant enzyme activities of Angora buck (Capra hircus ancryrensis) sperm following cryopreservation. Ejaculates collected from three Angora bucks were evaluated and pooled at 37 degrees C. Semen samples, which were diluted with a Tris-based extender containing the additives raffinose (2.5, 5, 10mM) and methionine (2.5, 5, 10mM) and an extender containing no antioxidants (control), were cooled to 5 degrees C and frozen in 0.25 ml French straws. Frozen straws were thawed individually at 37 degrees C for 20s in a water bath for evaluation. The freezing extender supplemented with 2.5 and 5mM methionine led to higher percentages of CASA motility (63.6+/-7.0; 63.4+/-3.1%, respectively), in comparison to the controls (P<0.01) following the freeze-thawing process. The addition of antioxidants did not provide any significant effect on the percentages of post-thaw subjective and CASA progressive motilities as well as sperm motion characteristics (VSL and VCL), compared to the control groups (P>0.05). The freezing extender with raffinose (5 and 10mM) and methionine at three different doses (2.5, 5 and 10mM) led to lower percentages of acrosome abnormalities, in comparison to the controls (P<0.001). In the comet test, raffinose (5 and 10mM) and methionine (10mM) gave scores lower than those of the controls, and thereby reduced DNA damage (P<0.05). Malondialdehyde formation was found to be lower (1.8+/-0.1 nmol/L) in the group of 5mM raffinose, compared to the controls following the freeze-thawing process (P<0.01). The additives did not show any effectiveness on the maintenance of SOD, GSH-PX and GSH activities, when compared to the controls (P>0.05). In conclusion, methionine and raffinose play a cryoprotective role against sperm CASA motility, acrosome abnormality and DNA damage. Raffinose 5mM exhibited antioxidative properties, decreasing MDA levels. Further studies are required to obtain more concrete results on the characterization of microscopic parameters and antioxidant activities in cryopreserved goat sperm with different additives.