Iron levels in the human brain: A post-mortem study of anatomical region differences and age-related changes

The link between brain iron homeostasis and neurodegenerative disease has been the subject of extensive research. There is increasing evidence of iron accumulation during ageing, and altered iron levels in some specific brain regions in neurodegenerative disease patients have been reported.Using graphite furnace atomic absorption spectrometry after microwave-assisted acid digestion of the samples, iron levels were determined in 14 different areas of the human brain [frontal cortex, superior and middle temporal, caudate nucleus, putamen, globus pallidus, cingulated gyrus, hippocampus, inferior parietal lobule, visual cortex of the occipital lobe, midbrain, pons (locus coeruleus), medulla and cerebellum (dentate nucleus)] of n = 42 adult individuals (71 ± 12 years old, range: 53–101 years old) with no known history or evidence of neurodegenerative, neurological or psychiatric disorders.It was found that the iron distribution in the adult human brain is quite heterogeneous. The highest levels were found in the putamen (mean ± SD, range: 855 ± 295 μg/g, 304–1628 μg/g) and globus pallidus (739 ± 390 μg/g, 225–1870 μg/g), and the lowest levels were observed in the pons (98 ± 43 μg/g, 11–253 μg/g) and medulla (56 ± 25 μg/g, 13–115 μg/g).Globally, iron levels proved to be age-related. The positive correlation between iron levels and age was most significant in the basal ganglia (caudate nucleus, putamen and globus pallidus).Compared with the age-matched control group, altered iron levels were observed in specific brain areas of one Parkinson's disease patient (the basal ganglia) and two Alzheimer's disease patients (the hippocampus).     

The ovulatory and LH responses to the male effect in dominant and subordinate goats

The objective of this study was to evaluate the LH and ovulatory response of dominant and subordinate dairy does following the introduction of males. A behavioral study was carried out to determine the individual success index (SI) of 35 anestrous does according to their ability to dominate other females. The 8 highest-ranking (highest SI) and the 8 lowest-ranking does (lowest SI) in terms of dominance were separated from the rest of the herd and placed together in the same pen, where a male was later introduced. Blood samples for plasma LH determinations were obtained from the 16 females at 30 min intervals from 0 to 6, 12 to 18 and 33 to 39 h after the introduction of the male, using an intravenous catheter. After day 8, plasma progesterone was also measured daily in order to determine the occurrence of ovulation. None of the does showed LH pulses during the sampling period prior to the introduction of the male. The dominant does showed significantly (p < 0.05) more LH pulses (2.0 ± 0.18 ng/ml vs. 1.2 ± 0.25 ng/ml) and a higher mean plasma LH concentration (0.25 ± 0.03 ng/ml vs. 0.14 ± 0.03 ng/ml) than the subordinate goats during the first 6 h of exposure to the male. There were no differences in LH pulsatility or concentrations at other times. The frequency of goats that ovulated tended to be greater in high-ranked than low-ranked does (87% vs. 37%). It could be concluded that the immediate LH response to the presence of the male (number of LH pulses) is higher in the dominant, compared to the subordinate goats, and this is associated with a higher number of dominant goats ovulating in response to the male effect.     

Serum progesterone concentrations,follicular development and time of ovulation using a new progesterone releasing device (DICO®) in sheep

Four experiments were conducted to evaluate the effectiveness of a new controlled drug releasing device containing 0.3 g progesterone (DICO^®) on ovarian control in sheep. In experiment 1, serum progesterone concentrations induced by a 14 days treatment of DICO^® (n = 9) and CIDR-G^® (n = 9) were compared in ovariectomized ewes. Both devices induced similar responses and no differences were recorded. In experiment 2, the onset of oestrus and the time of ovulation obtained after 14 days treatment with DICO^® (n = 8) and CIDR-G^® (n = 7) were compared in cyclic ewes. Both devices induced oestrus and ovulation in all of the ewes. The onset of oestrus (34.5 ± 2.8 and 30.0 ± 7.7 h), the time of ovulation (60.0 ± 9.1 and 54.9 ± 6.4 h), the ovulation rate (1.3 ± 0.5 and 1.4 ± 0.5), the follicular diameter at ovulation (7.0 ± 0.8 and 7.3 ± 1.1 mm), and the lifespan of the ovulatory follicles (8.6 ± 2.2 and 10.0 ± 2.9 days) were similar for the DICO^® and CIDR-G^® devices, respectively. In Experiment 3, the re-utilization of DICO^® devices inserted for 6 days (i.e. short-term protocol) was evaluated in ovariectomized ewes. The females received a re-used (previously used for 6 days; n = 11) or a new DICO^® (n = 11) for a period of 6 days. The re-used DICO^® devices induced a lower serum progesterone concentration than the new devices (P < 0.05). However, the re-used DICO^® device maintained serum progesterone concentrations above 7.1 nmol/L (i.e. >2 ng/ml) throughout treatment. In Experiment 4, the administration of eCG treatment at DICO^® withdrawal was evaluated in cyclic ewes. The short-term protocol using DICO^® devices for 6 days was applied with (n = 8) or without (n = 7) 300 IU eCG at the time of device withdrawal. The administration of eCG advanced ovarian follicular development, synchronizing the onset of oestrus at 36 h and the time of ovulation at 60 h from device withdrawal. In conclusion, data from these experiments show the use of DICO^® or CIDR-G^® devices containing 0.3 g of progesterone to have a similar efficiency in controlling serum progesterone concentrations, follicular development and the time of ovulation in sheep. The re-use of the devices, associated with the short-term protocol for 6 days is possible, although further studies on induced fertility rates are warranted.     

Postpartum ovarian activity of Santa Inês lactating ewes fed diets containing soybean hulls as a replacement for coastcross (Cynodon sp.) hay

The Santa Inês, a Brazilian hair sheep, has a non-seasonal breeding activity. Data regarding the duration of the postpartum anestrous period in Santa Inês lactating ewes is lacking and the objective of this trial was to determine the effects of replacing neutral detergent fibre (NDF) provided by coastcross (Cynodon sp.) hay with NDF contained in soybean hulls (SH) on the postpartum ovarian activity—as measured by the serum progesterone (P₄) concentration. Fifty-six lactating ewes (body weight 56.1 ± 6.8 kg) were individually penned and used in a randomized complete block design with 14 blocks and four treatments. The SH NDF replaced 33 (SH33), 67 (SH67), or 100% (SH100) of the NDF contributed by coastcross hay in the control diet (SH0). This resulted in a SH inclusion at rates of 0, 25, 54, and 85% of the dietary dry matter (DM). Blood samples were collected twice weekly from the 14th to 84th day postpartum and the serum P₄ concentrations determined by radioimmunoassay (RIA). It was estimated that the 1st postpartum ovulation occurred 6 days before the date that a serum P₄ ≥ 1 ng/ml concentration was first recorded. The mean body condition score (BCS; 0–5 scale) was 3.0 ± 0.19 on day 14 postpartum and the mean BCS at day 56 postpartum increased linearly (P < 0.01) with the inclusion levels of SH (3.09, 3.24, 3.34, and 3.36, respectively). Treatments did not differ significantly in the induction of postpartum days to the resumption of ovarian luteal activity (34.1 ± 15.3 days postpartum). On days 25, 50, and 75 postpartum 36, 80, and 100% of the ewes had resumed ovarian activity, respectively. Non-esterified fatty acid concentration decreased quadratically (P < 0.05) with the SH inclusion, with values of 0.323, 0.244, 0.204, and 0.216 mequiv./l for the SH0, SH33, SH67, and SH100 treatments being recorded, respectively. Replacement of the NDF provided by coastcross hay with the NDF from the SH did not influence the duration of the postpartum anestrous period in Santa Inês lactating ewes. Considering a 150-day gestation period and the 34 days postpartum anestrous demonstrated in the present study, the current production system of a lambing interval of 8 months (3 lambing events in 2 years) may not be optimizing the production potential and a system in which the lambing interval is shortened by at least 1 month may be feasible.     

Comparison of two methods of synchronization of estrus in brown brocket deer (Mazama gouazoubira)

This study aimed to establish a protocol for synchronization of estrus in brown brocket deer (Mazama gouazoubira). Two groups of hinds (n = 3) were submitted to two different protocols: Treatment 1 received an intravaginal progesterone (CIDR^®) device for 8 days, followed by 265 μg injection of cloprostenol at the time of removal; and Treatment 2 received two injections of 265 μg of cloprostenol 11 days apart. After 30 days, each group of three hinds received the other treatment. Treatment efficacy was evaluated by reproductive behavior, fecal progestin and estrogen concentration and the observation of CL by laparoscopy 6 days after the end of estrus. All the hinds (100%) had estrous behavior upon the completion of treatment, but a significant difference occurred between the time of onset, 70.5 ± 5.0 h for Treatment 1 and 52.3 ± 5.6 h for Treatment 2. The mean estrus duration time (34.7 ± 4.50 and 37.0 ± 8.11 h), ovulation rates (5/6 and 4/6), mean CL size (4.85 ± 0.74 and 3.21 ± 0.19 mm) and mean fecal progestin concentration at 6 days after the end of estrus (865.53 ± 76.59 and 1073.35 ± 106.82 ng/g feces) were not significantly different between treatments. There was no difference in fecal estrogen concentrations throughout the treatment and the greatest values of the estrogen:progestin ratio coincided with estrous behavior. Although fertility was not evaluated directly, both treatments were effective in synchronizing estrus in the species M. gouazoubira, with the formation of functional corpora lutea.     

Effect of anti-oxidants and oxidative stress parameters on ram semen after the freeze–thawing process

Oxidative damage to sperm resulting from reactive oxygen species generated by the cellular components of semen is one of the main causes for the decline in motility and fertility of sperm during the freeze–thawing process. The aim of this study was thus to determine the effects of anti-oxidants on standard semen parameters, lipid peroxidation (LPO) and anti-oxidant activities after the freeze–thawing of ram semen. Ejaculates collected from four Akkaraman rams, were pooled and evaluated at 33 °C. Semen samples were diluted in a Tris-based extender containing the anti-oxidants glutathione (GSH) (5 mM), oxidized glutathione (GSSG) (5 mM) or cysteine (5 mM) and an extender containing no anti-oxidants (control), cooled to 5 °C and frozen in 0.25 ml French straws. Frozen straws were thawed individually for 20 s in a water bath (37 °C) for microscopic evaluation. The use of an extender supplemented with cysteine led to the highest (P < 0.01) post-thaw motility (61.0 ± 1.9%), compared to the other treatment groups. No significant differences were observed in viability, acrosome damage and total abnormalities, and following the hypo-osmotic swelling test (HOST), following supplementation with anti-oxidants after the thawing of the semen. Following the thawing process, the levels of malondialdehyde (MDA) did not change with the addition of anti-oxidants, compared to the control. The GSH level and glutathione peroxidase (GSH-PX) activity remained significantly higher upon the addition of GSH (3.33 ± 0.14 nmol/ml and 22.02 ± 1.27 IU/g protein) and GSSG (3.24 ± 0.08 nmol/ml and 20.17 ± 3.38 IU/g protein) compared to the other treatment (P < 0.001) groups. Only cysteine significantly elevated the activity of catalase (CAT, 842.40 ± 90.42 kU/l) following the freeze–thawing process. The Vitamin E (VitE) level was significantly higher, when compared to GSSG, cysteine and the control, when GSH (4.21 ± 0.20 mg/dl) was added to the freezing extender (P < 0.001). It could be concluded that future efforts aimed on improving the efficiency of cryopreservation of ram sperm should concentrate on the use of anti-oxidant additives. The results obtained provide a new approach to the cryopreservation of ram semen, and could positively contribute to intensive sheep production.     

Levels of l-ascorbic acid and cadmium in the saphenous vein of patients with coronary artery disease are negatively correlated

BackgroundThe aim of this study was the simultaneous determination of levels of cadmium and l-ascorbic Acid (AA) in human saphenous vein (SV) used in coronary artery bypass grafting (CABG) and check whether there is a relationship between these levels.MethodsHuman SV were collected from 40 patients (20 men and 20 women; age, 40–75 years) at the time of routine coronary artery surgical revascularization. The concentration of cadmium in the tissue was determined according to the GF AAS—atomic absorption method. The concentration of AA was assayed in supernatant by FIA method with spectrophotometric detection.ResultsAA concentration (mean ± SD); men: 98,7 ± 13,18 μg/g tissue, women: 96,06 ± 11,98 μg/g tissue. Cadmium concentration(mean ± SD); men: 309 ± 103,71 ng/g tissue, women: 348,5 ± 255,71 ng/g tissue. Correlations among concentrations of AA and cadmium were insignificant negative in the group of men (Pearson r = −0,1504, p = 0,5269) and in the group women (Pearson r = −0339, p = 0144).ConclusionsNegative correlations among concentrations of AA and cadmium in human SV obtained in our study may indicate a protective effect of this vitamin in relation to toxic cadmium.     

Effects of estradiol and progesterone on circulating LH and FSH secretion,and ovarian antral follicle growth in anestrous ewes

In the ewe, ovarian antral follicles emerge or grow in a wave-like pattern and each wave is preceded by a peak in the serum FSH level. The purpose of the current study was to investigate whether in anestrous Western White Face ewes, a combination of progesterone and estradiol affects the circulating FSH peak secretion and the number of small ovarian follicles. Five ewes were treated with subcutaneous silastic rubber implants (10 cm × 0.47 cm), containing 10% estradiol-17β w/w (controls) and 5 ewes were treated with the same estradiol implant, along with subcutaneous implants (11 cm × 0.48 cm) containing 10% progesterone w/w for 12 days. Daily transrectal ovarian ultrasonography and blood sampling was performed from 5 days before, to 9 days after the period of implantation. Blood samples were also taken every 12 min for a 6 h period on day −2, 6 and 13 prior to or after implant insertion (day 0, day of implant insertion). Pulsatility in the serum LH levels was eliminated by the implants (P < 0.05). During the implantation period, the serum FSH peak amplitude was lower in ewes treated with implants releasing estradiol and progesterone, compared to ewes treated with implants releasing only estradiol (P < 0.05). No follicular waves emerged during implant treatment in both groups (P < 0.05) and the number of serum FSH peaks did not differ during implantation, compared to before implantation. During the implantation period, the number of small follicles did not differ in ewes with implants releasing estradiol and progesterone, compared to ewes treated with implants releasing only estradiol. To conclude, supra-physiological concentrations of estradiol completely eliminated the serum LH pulsatality and suppressed the follicular wave emergence, while the FSH secretory peaks that preceded the follicular waves were not affected. Supra-physiological concentrations of estradiol-17β with physiological concentrations of progesterone decreased the serum FSH peak amplitude, eliminated the serum LH pulses, but did not decrease the size of the small follicle pool in anestrous ewes.     

Active immunization against inhibin-based peptides to increase ovulation rate in non-prolific Malpura ewes

The aim of this study was to explore the possibility of increasing the ovulation rate of Malpura, a non-prolific tropical breed of sheep by immunization against inhibin-based peptide immunogens. Ewes were divided into three groups (n = 5 each) and actively immunized against the synthetic peptides from the α_C [bIα(1–29)-Tyr³⁰] or α_N [bI-₄₃-Tyr¹⁵²(153–167)Cys¹⁶⁸] area of the bovine inhibin α-subunit conjugated to ovalbumin or against ovalbumin (control). Each ewe received a primary immunization of 400 μg immunogen and 3 boosters, each of 200 μg immunogen at 4-week intervals. Estrus was synchronized using a double PGF₂α injection schedule and laparoscopy was performed after each estrus to determine the ovulation response. Immunization against both the peptides did not affect the interval from PGF treatment to the onset of estrus, the duration of estrus and the number of large unovulated follicles. In contrast to the complete absence of multiple ovulations in the controls, all the ewes immunized against α_C or α_N peptides showed multiple ovulations (range 2–7) in all the three estrous cycles evaluated, except for one ewe immunized against the α_N peptide, which exhibited multiple ovulations in only 1 out of the 3 estrous cycles. Compared to that of the controls (1.0 ± 0.9, 1.0 ± 0.0 and 0.6 ± 0.2, respectively), the mean ovulation rate was higher (P < 0.01) in the ewes immunized against the αC (4.8 ± 1.02, 5.0 ± 1.05 and 5.0 ± 0.45, respectively) or against αN (4.5 ± 1.19, 2.5 ± 0.87 and 2.7 ± 0.75, respectively, P < 0.05) peptide in estrous cycles numbers 1, 2 and 3. These results show that active immunization against inhibin-based peptide immunogens is effective in increasing ovulation rate in Malpura, a non-prolific breed of sheep and that it may be an alternative to conventional superovulation regimes.     

Rapid determination of 25-hydroxy vitamin D3 in swine tissue using an isotope dilution HPLC-MS assay

A rapid method for quantification of 25-hydroxy vitamin D3 in different swine tissues based on isotope dilution HPLC-MS has been developed and validated. Six times deuterated analyte is used as internal standard. The method is fast and can be performed with only 1 g sample. Sample preparation for kidney, liver, muscle and spleen requires only homogenisation and extraction with methanol. An additional enzymatic digest is required for skin, and clean-up of the extract by solid-phase extraction (SPE) is used for adipose tissue and skin. The lower limit of detection varies from 1 ng/g (muscle) to 5 ng/g (adipose and skin). The method has been successfully applied to various tissue samples of pigs fed for 119 days either 2000 IU of vitamin D3 or 50 μg of 25-hydroxy vitamin D3 per kg feed. For animals ingesting 25-OH-D3 supplements the highest tissue contents were observed in the skin (24.8 ± 3.5 ng/g), followed by kidney (14.2 ± 1.5 ng/g), liver and muscle (5.7 ± 0.6 ng/g). The 25-OH-D3 content in the skin was significantly higher in animals ingesting 2000 IU/kg of vitamin D3 (39.5 ± 13.4 ng/g). Levels in selected tissues of some animals were below the lower limit of quantification. No measurable amounts of 25-OH-D3 were found in spleen, abdominal fat and subcutaneous fat of the animals of both groups as well as in the liver, kidney and muscle of the animals ingesting 2000 IU/kg of vitamin D3.     

The effect of dietary supplementation with calcium salts of long chain fatty acids and/or l-carnitine on ovarian activity of Rahmani ewes

This study investigated the effect of dietary supplementation with calcium salts of long chain fatty acids with or without of l-carnitine on ovarian activity using 24 Rahmani ewes randomly allocated to four treatments. Control animals (n = 6) were fed a basal diet of hay (64.2%) and barley grain (35.0%) plus minerals and vitamins (0.8%). Ewes on the three treatments received the same basal diet supplemented with calcium salts of long chain fatty acids (CSFA) at 3% of the basal diet dry matter intake (1.4 kg/ewe/d); 250 ppm l-carnitine (LC); or both these supplements (CSFA + LC). All use exhibited natural estrus on one or two occasions and were weighed at the start and the end of the study as well as body condition score was assessed at the end of study. All ewes were then synchronised for estrus using intravaginal sponges for 12 d prior to the start of the nutritional treatments and three weeks after the nutritional treatments began. The nutritional treatments were imposed for a total of 8 weeks. Blood samples were collected prior to the start of treatments and every two weeks thereafter except after sponge removal of first and second synchronisation where the blood samples were collected daily for progesterone assay. The results revealed that Rahmani ewes received basal diet (control) and l-carnitine had significantly decrease final body weight and body condition score (36.3 ± 0.4; 36.8 ± 0.3; 2.2 ± 0.04; 2.1 ± 0.05; p < 0.05, respectively) than those on CSFA and CSFA + LC (38.6 ± 0.9; 39.5 ± 0.6; 3.3 ± 0.07; 3.4 ± 0.06; respectively). At the second ultrasound examination, the control animals had significantly fewer total follicles (7.3 ± 0.8; p < 0.05) than those on the CSFA (8.4 ± 0.8), l-carnitine (8.7 ± 1.5) and CSFA + LC (8.0 ± 0.6) treatments. The increased numbers occurred in the medium and large categories of follicles. In addition, the ovulation rates were significantly lower (p < 0.05) for control (1.3 ± 0.2) and l-carnitine (1.5 ± 0.00) than for CSFA (2.5 ± 0.3) and CSFA + LC (2.3 ± 0.2). Furthermore, serum progesterone concentrations had risen and were significantly higher (p < 0.05) for CSFA (2.5 ± 0.3 ng/ml) and CSFA + LC (2.7 ± 0.1 ng/ml) than for control (1.1 ± 0.7 ng/ml) and l-carnitine (1.5 ± 0.4 ng/ml). It was concluded that supplementation of the basal diet with l-carnitine alone did not improve performance of ewes or the ovarian response. However, the addition of calcium salts of long chain fatty acids to the basal diet alone or in combination with l-carnitine significantly improved the number and size of ovarian preovulatory follicles, and the ovulation rate of Rahmani ewes. Further evidence was required to study their influence on follicular atresia.     

Liquid chromatography-tandem mass spectrometry method for determination of the pyridinium aldoxime 4-PAO in brain,liver, lung,and kidney

A liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was validated and applied to the in vitro determination of 4-[(hydroxyimino)methyl]-1-octylpyridinium cation (4-PAO), which can penetrate the blood–brain barrier and reactivate acetylcholinesterase (AChE) inhibited by alkylphosphonate in the brain, liver, lung, and kidney. The limit of detection (LOD) was 0.235 μg cation/g wet weight, and the quantification range and linearity of the calibration curve extended over a range of 0.470–941 μg cation/g wet weight. For the proof of applicability, when 4-PAO was administrated intravenously via the rat tail vein at 10% LD₅₀, we were able to quantify the 4-PAO concentration in the tissues: brain 7.60 ± 1.32 μg cation/g wet weight (mean ± SD, n = 5), liver 26.8 ± 2.82 μg cation/g, lung 76.4 ± 24.9 μg cation/g, and kidney 638 ± 266 μg cation/g. In addition, the methods for 4-[(hydroxyimino)methyl]-1-decylpyridinium bromide (4-PAD) and 4-[(hydroxyimino)methyl]-1-(2-phenylethyl) pyridinium bromide (4-PAPE) were partly validated referring to the findings of the 4-PAO full validation. Thus, the LC-MS/MS method described in this study can be useful for quantification of pyridinium aldoxime methiodide (PAM)-type oximes in biological samples.     

Signals from fat after injury: Plasma adipokines and ghrelin concentrations in the severely burned

IntroductionHypermetabolism is universal in the severely burned and is characterized by catabolism of lean mass and body fat with associated insulin resistance. Adipokines are likely to play a role in these changes but have not been identified to date in burn patients.MethodsFrom a single burn ICU, 17 burn patients with an expected stay >14 days were studied. Study period began within 14 days of admission. Over 7 days, plasma samples were collected for measurement of leptin, adiponectin, resistin, ghrelin, insulin, and cortisol by ELISA. For comparison, samples from 15 healthy controls of similar age, BMI, and blood glucose were obtained.ResultsMean age was 33 ± 17 years and BMI 26 ± 3.4. Average burn size was 45 ± 20% TBSA and ISS 32 ± 10 with 72% having inhalation injury; in-hospital mortality was 29%. Estimated energy needs were 3626 ± 710 kcal, of which 84 ± 21% were met by enteral feeding with intensive insulin treatment (glucose 80–110 mg/ml). Using the homeostasis model assessment of insulin resistance, burned subjects were more resistant than controls (17 ± 11.3 and 8 ± 10.0). Insulin levels were elevated (57 ± 35.6 μU/ml in burned subject vs. 26 ± 31.1 μU/ml in controls), and cortisol concentrations increased (50 ± 41.2 μg/dl vs. 12 ± 3.9 μg/dl). These traditional hormone changes were associated with increased resistin (16.6 ± 5.5 ng/ml vs. 3.8 ± 0.9 ng/ml) and decreased leptin (8.8 ± 8.9 ng/ml vs. 19.4 ± 23.5 ng/ml), adiponectin (9 ± 3.5 ng/ml vs. 17 ± 10.2 ng/ml), and ghrelin (0.37 ± 0.14 ng/ml vs.0.56 ± 0.26 ng/ml).ConclusionPatients with burns, who are characteristically hypermetabolic with hypercortisolism and insulin resistant, have significant changes in adipokine levels that appear independent of the magnitude of initial injury or metabolic derangement. In addition, suppression of ghrelin in the presence of decreased leptin and adiponectin levels in combination with increased insulin and resistin levels represent unexpected changes in the metabolic milieu of the injured patient possibly due to dramatic activation of inflammatory pathways, indicating strategies for treatment.     

The effect of glucose and maltose concentrations on pyruvate and ascorbate production,antioxidant enzyme activities and LPO levels in Fusarium equiseti

Changes in pyruvate and ascorbate production and antioxidant enzyme activities together with lipid peroxidation levels in Fusarium equiseti were investigated in relation to changes in the concentrations of glucose and maltose as carbon sources in the range of 5–25 g/l in Armstrong Fusarium Medium (AFM). The highest pyruvate concentration obtained at 20 g/l maltose was 67.5 ± 0.69 μg/ml while ascorbic acid reached a maximum value at 25 g/l glucose of 1866±26.1 μg/ml The maximum superoxide dismutas (SOD) activities related to increased pyruvate production were determined in AFM medium containing 20 g/l glucose as 41.49±0.65 and maltose as 61.12±0.8 IU/mg. Catalase (CAT) activity variations showed coherence with SOD activity in a medium containing maltose and reached 219.11±2.8 IU/mg while they were decreased with increasing glucose concentration. glutathione peroxidase (GSH-Px) activities in F. equiseti did not change significantly with glucose and maltose concentration and were determined to be 1.21±0.22 and 1.67±0.15 IU/mg, respectively. Minimum lipid peroxidation levels for each carbon source were determined in both 20 g/l maltose and glucose concentrations as 0.9 and 1.62 nmol MDA/g wet weight.     

Social ranking and plasma progesterone levels in goats

Progesterone is essential for maintaining pregnancy in goats, and embryonic losses may be a consequence of the reduction in circulating progesterone levels close to the time of implantation. Some evidence exists regarding social dominance affecting the plasma progesterone levels in several species—where dominant females conceive earlier. The objective of this research was to determine whether serum progesterone levels differ in goats of different social status. A behavioural study was conducted for 10 days in a herd of 57 does and an individual success index (SI) was calculated according to the result of aggressive interactions. Goats were classified as high (SI: 1–0.67), medium (SI: 0.66–0.34) and low-ranking (SI: 0.33–0.0). Ovulation was synchronized using two injections of prostaglandin 11 days apart, and the plasma progesterone levels determined daily for a period of 20 days. The area under the plasma progesterone curve during the entire study was greater in the high than in the medium and low-ranking does (96.2 ± 5.8, 79.5 ± 5.3 and 81.3 ± 5.3 ng/ml, respectively, P < 0.05). During days 11–17 following prostaglandin synchronization, the plasma progesterone levels were higher in the high-ranking (P < 0.05), compared to the low-ranking does. Plasma progesterone levels were significantly correlated with SI at days 14 and 15 (r = 0.26; P < 0.05). Results suggest a relationship between social ranking of goats and the plasma progesterone production from the corpus luteum and other possible sources.     

Menstrual Disorders and Androgen-Related Traits in Young Women with Type 1 Diabetes Mellitus: a Clinical Study

Objective: To investigate possible causes of menstrual disorders and androgen-related traits in young women with type 1 diabetes mellitus (T1DM).Methods: Fifty-three women with T1DM (duration 8.0 ± 5.6 years), 41 women with (polycystic ovary syndrome) PCOS, and 51 controls matched for age (19.4 ± 4.3 years vs. 21.2 ± 2.7 years vs. 20.8 ± 3.1 years; P>.05) and body mass index (BMI) (22.2 ± 2.7 kg/m² vs. 21.9 ± 2.0 kg/m² vs. 21.4 ± 1.9 kg/m²; P>.05) were prospectively recruited.Results: Two women (3.8%) in the T1DM group had not experienced menarche (at 15.5 and 16.6 years); of the rest, 23.5% had oligomenorrhea, 32.1% hirsutism, and 45.3% had acne. The age at menarche was delayed in the T1DM group compared to controls (12.7 ± 1.3 vs. 12.0 ± 1.0 years; P = .004), while no difference was observed with the polycystic ovary syndrome (PCOS) group (12.4 ± 1.2 years). There were no differences in total testosterone (0.43 ± 0.14 ng/mL vs. 0.39 ± 0.14 ng/mL; P>.05), dehydroepiandrosterone sulfate (DHEA-S) (269 ± 112 μg/dL vs. 238 ± 106 μg/dL; P>.05) or Δ4-androstenedione (2.4 ± 1.3 ng/mL vs. 1.9 ± 0.5 ng/mL; P>.05) concentrations between T1DM and controls. However, patients with T1DM had lower sex hormone binding globulin (SHBG) concentrations than controls (61 ± 17 nmol/L vs. 83 ± 18.1 nmol/L; P = .001), which were even lower in the PCOS group (39.5 ± 12.9 nmol/L; P = .001 compared with T1DM). The free androgen index (FAI) was higher in the PCOS group compared with both other groups (T1DM vs. PCOS vs. controls: 2.53 ± 0.54 vs. 7.88 ± 1.21 vs. 1.6 ± 0.68; P<.001). FAI was higher in patients with T1DM compared to controls as well (P = .038). There was no difference in DHEA-S concentrations between T1DM and PCOS patients (269 ± 112 μg/dL vs. 297 ± 100 μg/dL; P>.05).Conclusion: Menstrual disorders and androgen-related traits in young women with T1DM may be attributed to an increase in androgen bioavailability due to decreased SHBG concentrations.     

Production of glucaric acid from myo-inositol in engineered Pichia pastoris

A potential myo-inositol oxygenase (ppMIOX) was identified as a functional enzyme and a glucaric acid synthetic pathway was firstly constructed in Pichia pastoris. Coexpression of the native ppMIOX and the urinate dehydrogenase (Udh) from Pseudomonas putida KT2440 led to obvious accumulation of glucaric acid (90.46 ± 0.04 mg/L) from myo-inositol whereas no glucaric acid was detected from glucose. In comparison, coexpression of the heterologous mouse MIOX (mMIOX) and Udh resulted in higher titers of glucaric acid from glucose and myo-inositol, 107.19 ± 11.91 mg/L and 785.4 ± 1.41 mg/L, respectively. By applying a fusion expression strategy with flexible peptides, the mMIOX specific activity and the glucaric acid concentration were significantly increased. Using glucose and myo-inositol as carbon substrates, the production of glucaric acid was substantially enhanced to 6.61 ± 0.30 g/L in fed-batch cultures. To the best of our knowledge, this is the highest reported value to date.     

Phenotypic and genetic relationships between udder morphology and udder health in dairy ewes

The aim of this paper was studying the phenotypic and genetic relationships between udder morphology traits and udder health in dairy sheep. From 2000 to 2008 udder traits, somatic cell count (SCC) and clinical mastitis cases were recorded in an experimental flock consisting each year of around 900 ewes. A logistic regression was performed to evaluate the risk for a ewe of showing either a mastitis or at least 2 daily SCC records greater than 1 × 10⁶ cells/ml in one of its lactations as function of udder traits scored in 1st lactation. 1587 individual udder scores and 39,950 SCC daily records were used for the analysis. Secondly, genetic correlation between lactation mean of somatic cell score and udder traits were estimated by a REML method applied to four bi-trait animal models, using data from 2251 primiparous ewes. Logistic regression results indicated that the risk of mastitis or high SCC values during the productive life increased as the cistern height increased and the degree of udder suspension and udder depth decreased. This suggests that the appraisal of the udder is a useful tool for culling decision aimed at increasing the sanitary status of the flock. The genetic correlation between lactation SCS and udder traits were favourable for udder depth (−0.50 ± 0.12), teat placement (0.39 ± 0.011) and degree of udder suspension (−0.42 ± 0.011) and closed to zero for the degree of separation of the two halves. Thus selection for udder morphology, already implemented in some dairy sheep breeds with the aim of improving machine milkability, will lead to a favourable correlated genetic response on SCC.     

Effects of lambing season,sex and birth type on growth performance in Norduz lambs

The aim of this study was to evaluate effects of two production systems (spring and winter), sex and birth type on growth performance in Norduz lambs. Data were collected using 103 and 140 lambs born during the winter and spring, respectively. Lambs born in the winter were heavier (P < 0.01) than those born in the spring at birth, 90 and 180 days of age by 0.5, 1.6 and 1.7 kg, respectively. Ram lambs were heavier (P < 0.01) than ewe lambs at birth, 90 and 180 days of age by 0.5, 1.0 and 2.3 kg, respectively, while lambs born as singles were heavier (P < 0.01) than lambs born as twins at birth, 90 and 180 days of age by 0.9, 1.6 and 2.3 kg, respectively. Lambs born in the winter recorded a higher (P < 0.01) pre-weaning ADG (15 g/day) than spring-born lambs, while ram lambs recorded a higher (P < 0.01) pre-weaning ADG (11 g/day) than ewe lambs. The effect of birth type was not significant on pre-weaning ADG. The influence of lambing season and sex were also not significant on post-weaning ADG. Lambs born as singles recorded a higher (P < 0.01) post-weaning ADG (16 g/day) than lambs born as twins. Results suggest the winter season to give rise to heavier lambs at weaning and post-weaning and suggest lambs born in different seasons to have distinctly different growth patterns. The data also confirm that the influence of sex and type of birth on growth to be very important and to be take into account sheep production.     

The role of a cleft upper lip of alpacas in foraging extremely short grasses evaluated by grazing impulse

The role of a cleft upper lip of alpaca (Lama pacos) in foraging short pastures was investigated using biting forces and associated impulses in animal grazing. Three Merino wethers and three castrated alpacas were used. Ten (10L), 20 (20L), 30 (30L), 40 (40L) or 50 (50L) leaves of orchardgrass (Dactylis glomerata) per load cell were offered to animals, and three-directional biting forces were digitally recorded at 5 s⁻¹⁰⁰⁰. From the total biting force/time curve, grazing impulse was calculated, equivalent to the area surrounded by the curve. The grand mean of the number of grazed leaves per bite was 9.8 ± 0.53 in alpaca and 17.9 ± 1.31 in sheep. Remaining leaf length after grazing trial was significantly lower in alpaca than in sheep (11.9 ± 0.19 mm versus 18.5 ± 0.41 mm). Alpaca grazed leaves with significantly lower mean biting force (7.0 ± 0.69 N versus 20.0 ± 1.80 N) and significantly shorter duration time per one biting force (0.11 ± 0.005 s versus 0.18 ± 0.19 s) than sheep. The grand mean of sum of grazing impulse was lower (P < 0.002) in alpaca (2.8 ± 0.42 N s) than in sheep (9.4 ± 1.95 N s). Grazed DM weight increased with increasing leaf densities, but there was no difference between alpaca and sheep. The ratio of DM intake to grazing impulse was significantly higher in alpaca than in sheep (0.18 ± 0.016 g DM/N s versus 0.08 ± 0.010 g DM/N s). The grazing strategy in alpaca seems to be an adaptation for foraging extremely short grasses. Alpacas may push aside cleft upper lips when trying to grasp short leaves, resulting in keeping incisors at lower insertion position near the ground surface.