The effect of heat stress on performance, fertility, and adipokines involved in regulating systemic immune response during lipolysis of early lactating dairy cows.

The aim of this study was to assess the potential effect of heat stress on dairy cow productivity, fertility, and biochemical blood indices during the early lactation stage in a temperate climate. Additionally, the study aimed to determine the role of leptin and adiponectin in regulating the immune response accompanying lipolysis after calving in dairy cows. The study included 100 clinically healthy Polish Holstein-Friesian dairy cows selected based on parity and 305 d of milk yield from 5 commercial farms with similar herd management and housing systems. Prospective cohort data were recorded from calving day until 150 d in milk, and microclimate loggers installed inside the barns were used to record temperature and relative humidity data to calculate daily temperature-humidity index (THI) on the calving day, through +7, +14, and +21 d during early lactation. Additionally, monthly productive performance parameters such as milk yield, chemical composition, fatty acids composition, and fertility indices were analyzed. Results showed that the THI from calving day through +7, +14, and +21 d during early lactation was negatively associated with fertility parameters such as delayed first estrus postpartum and an elongated calving interval, respectively, by 29, 27, 25, and 16 d. Furthermore, an increase in THI value during early lactation was associated with an elongated artificially inseminated service period, days open, and intercalving period. Increasing THI from calving day (0 d) through +7, +14, and up to +21 d during early lactation was also linked to decreased milk yield by 3.20, 4.10, 5.60, and 5.60 kg, respectively. The study also found that heat stress during early lactation was associated with a lower body condition score in dairy cows and higher concentrations of leptin, nonesterified fatty acids, and ß-hydroxybutyrate, accompanied by a drastic reduction in adipose tissue-secreted adiponectin levels after calving. Additionally, heat stress-induced lipolysis in adipose tissue caused an inflammatory response that increased biochemical blood indices associated with immune responses such as cytokines, acute phase proteins, and heat shock protein. These findings suggest that exposing dairy cows to heat stress during early lactation can negatively affect their productive performance, fertility, and biochemical blood indices in subsequent lactations. Thus, farm management changes should be implemented during early lactation to mitigate the negative consequences of heat stress occurrence.

Short- and long-term effects of initial serum total protein, average starter feed intake during the last week of the preweaning period, and rearing body gain on primiparous dairy heifers' performance.

The aim of this study was to evaluate the short- and long-term effects of initial serum total protein (STP) concentration, average starter feed intake (SI) during the last week of the preweaning period, and average daily gain (ADG) on the growth, fertility, and performance of Holstein heifers during their first lactation. Eighty-four female Holstein dairy calves were weaned at d 56 of age and then the study continued until the end of the first lactation. Growth performance, including body weight, ADG, withers height, and its change were analyzed monthly from 3 to 450 d of life, and reproduction data and performance in the first lactation of primiparous dairy heifers over a 4-yr period (2015 to 2019) were recorded. In the first 24 h of life, calves received 4 L of colostrum (<2 h and <12 h after birth); on d 2 and 3: 2 feedings/d of 2 L of transition milk; from d 4 to 49: 6 L/d of milk replacer (150 g of powder/L as-fed) in 3 feedings; and from d 50 to 56: 2 L/d of milk replacer in 1 feeding. The calves were fed pelleted starter feed from d 4 to 56, and after that from 8 wk until 3 mo of age, a dry total mixed ration with an 85:15 ratio of weaning pelleted starter to straw. From 3 to 7 mo and from 8 mo of age to calving, the total mixed ration contained 16.9% and ~14.0% crude protein, respectively, on a dry matter basis and ~2.40 Mcal of metabolizable energy/kg on a dry matter basis. The results of the current study showed that the initial STP concentration of primiparous dairy heifers was associated with improved growth performance, especially greater body weight and withers height. In addition, with increasing levels of initial STP concentration, age at first estrus, artificial insemination (AI) service, pregnancy, and calving was decreased by 16, 18, 25, and 25 d, respectively. Initial STP concentration was positively correlated with milk production and increased total milk yield and yield of energy-corrected milk by about 1,558 kg and 1,149 kg during first lactation. Calves with higher average starter feed intake during the last week of the preweaning period had better growth performance, which in turn was positively associated with fertility parameters, accelerated first estrus (by 17 d), and reduced age at AI service (by 13 d). Preweaning ADG was favorably associated with fertility performance of heifers, with faster occurrence of first estrus and a reduction in age at AI service, pregnancy, and calving. Also, increasing preweaning ADG increased milk yield, energy-corrected milk, and 4% fat-corrected milk at 305 DIM by about 829, 754, and 763 kg at first lactation of primiparous heifers. These results indicate that in the rearing period, particular attention should be paid to the initial STP concentration, average SI during the last week of this period, and rearing ADG to increase growth, fertility, and performance in the first lactation of primiparous dairy heifers.

Photoinduced transport in an H64Q neuroglobin antidote for carbon monoxide poisoning.

Carbon monoxide (CO) is a leading cause of poisoning deaths worldwide, without available antidotal therapy. Recently, a potential treatment for CO poisoning was introduced, based on binding of CO by neuroglobin (Ngb) with a mutated distal histidine (H64Q). Here, we present an atomistic mechanism of CO trapping by H64Q Ngb revealed by nonadiabatic molecular dynamics. We focused on CO photodissociation and recombination of CO to wild type (WT) and H64Q Ngb. Our results demonstrate that the distribution of CO within the proteins differs substantially due to rearrangement of amino acids surrounding the distal heme pocket. This leads to the decrease of the distal pocket volume in H64Q Ngb in comparison to WT Ngb, trapping migrating CO molecules in the distal pocket. We show that the mutation implicates the shortening of the time scale of CO geminate recombination, making H64Q Ngb 2.7 times more frequent binder than WT Ngb.

Subacute ruminal acidosis affects fermentation and endotoxin concentration in the rumen and relative expression of the CD14/TLR4/MD2 genes involved in lipopolysaccharide systemic immune response in dairy cows.

The first objective of this study was to investigate the effects of subacute ruminal acidosis (SARA) on fermentation, ruminal free lipopolysaccharides (LPS), and expression of the cluster of differentiation 14 (CD14), toll-like receptor 4 (TLR4), and myeloid differentiation protein 2 (MD2) complex in white blood cells involved in the systemic immune response in dairy cows. The second objective was a study of whether increased expression of the LPS receptor complex led to increases in the concentrations of plasma high-density lipoprotein (HDL) and serum Ca. Three hundred five dairy cows located in 13 Polish high-yielding dairy commercial farms were selected according to their days in milk (40-150 d; average = 75), 305-d milk yield (10,070-12,041 kg; average = 10,940), and number of lactations (primiparous, n = 139 and multiparous, n = 166). Next, the herds were segregated into 3 groups based on the percentages of cows with an assigned value of ruminal fluid pH: SARA-positive, SARA-risk, and SARA-negative herds. Moreover, 305 selected dairy cows were divided according to the classification based on ruminal fluid pH into 3 groups as healthy (pH >5.81), risk (pH 5.8-5.6) and acidotic cows (pH <5.6). Rumen fluid samples were collected via rumenocentesis. In the AC group, we recorded higher concentrations of ruminal free LPS [4.57 Log10 endotoxin units (EU)/mL; 42,206 EU/mL] compared with the healthy group (4.48 Log10 EU/mL; 34,179 EU/mL). Similarly, the concentration of ruminal free LPS was higher in SARA-positive herds (4.60 Log10 EU/mL; 43,000 EU/mL) compared with SARA-negative herds (4.47 Log10 EU/mL; 32,225 EU/mL). The relative mRNA abundance of genes associated with the function of LPS receptors, such as CD14, TLR4, and MD2, in white blood cells differed between all experimental groups on both cow and herd levels. In the acidotic group, we recorded higher concentrations of HDL (78.16 vs. 68.32 mg/dL) and serum amyloid A (10.80 vs. 9.16 µg/mL) and lower concentrations of Ca (8.26 vs. 10.16 mg/dL) and haptoglobin (470.19 vs. 516.85 ng/mL) compared with the healthy group. Similar results were obtained in the SARA herd status analysis, but the concentration of lipopolysaccharide-binding protein differed statistically. Moreover, the pH of ruminal fluid was negatively correlated with relative mRNA abundance of genes such as CD14, TLR4, MD2, and concentrations of serum HDL and serum amyloid A, although positively correlated with serum Ca. The results indicated that decreases in ruminal fluid pH increased the release of free LPS into the rumen and stimulated the expression of the LPS receptor complex and immune response. Moreover, an increase in the expression of the LPS receptor led to higher concentrations of plasma HDL and lower serum Ca, which may be a protective mechanism against endotoxemia. However, the biological significance of these results needs to be investigated further in larger field trials.

Prevalence and consequence of subacute ruminal acidosis in Polish dairy herds.

The aim of this study was to estimate the prevalence of subacute ruminal acidosis (SARA) in Polish high-yielding dairy herds. Also, the relationships between the chemical composition of the diet and the feed particle size, ruminal pH and the occurrence of this metabolic disease and the fermentation profile were determined. Rumen fluid samples were obtained from a total of 213 cows from nine dairy herds (≥20 cows per herd) via rumenocentesis. Almost 14% (30/213) of cows as acidotic (pH<5.6) were found, which is indicative of SARA. Moreover, 44% of the herds were classified as SARA-positive and 56% as SARA-negative. Results of the current study suggested that the physically effective NDF (NDF>1.18 mm) -to-starch ratio could be better indices than peNDF>1.18 mm to preventing the occurrence of SARA, and their level should not be lower than 1.00. Also, linear negative relationships between rumen fluid pH and concentration of propionate, valerate and total VFA were determined. According to the herd's SARA status and rumen fluid biochemical indices, there were significant differences between the pH of rumen fluid (p ≤ 0.01), concentrations of acetate (p ≤ 0.05), propionate (p ≤ 0.05), n-butyrate (p ≤ 0.01), n-valerate (p ≤ 0.01), the sum of VFA (p ≤ 0.01) and ammonia (p ≤ 0.05) in SARA-positive herds compared to SARA-negative herds. The better understanding the strategy of ruminant nutrition to coordinate energy conversion and the role of the ruminal pH in regulating N-NH3 production, absorption through rumen mucosa, urea secretion, the more successfully we can utilize these processes with due recognition of animal needs and welfare, as well as prevention of SARA occurrence.

Relationship between pH of ruminal fluid during subacute ruminal acidosis and physiological response of the Polish Holstein-Friesian dairy cows.

The aim of the study was to investigate the effect of ruminal fluid pH depression on biochemical indices of blood, urine, feces, and milk, and to determine which of them may be helpful as a marker for the diagnosis of subacute ruminal acidosis (SARA). Ruminal fluid samples were obtained by rumenocentesis from 305 cows representing 13 commercial dairy herds. The herds were selected based on percentages of cows with an assigned value of ruminal fluid pH segregated into three groups as: SARA-positive herd, if at least 25% of the ruminal fluid samples indicated a pH < 5.6; SARA-risk herd, if less than 25% of ruminal fluid samples indicated a pH < 5.6, but at least 33% showed a pH ≤ 5.8; and SARA-negative herd, if less than 25% of the ruminal fluid samples indicated a pH < 5.6, but less than 33% exhibited a pH = 5.8. Moreover, the dairy cows were divided according to the ruminal fluid pH into three groups as follows: healthy cows (HC, pH>5.80, n = 196), risk cows (RC, pH 5.8 - 5.6, n = 51), and acidotic cows (AC, pH < 5.6, n = 58). Almost 19% (58/305) of the cows were classified as acidotic (pH < 5.6) and 46.2% of the herds as SARA-positive. In the AC group, higher concentrations of insulin-like growth factor-I (IGF-I), non-esterified fatty acids (NEFA), rectal temperature and lower blood pH, compared with those of the HC group, were recorded. Moreover, in the SARA-positive herds, higher concentrations of IGF-I and the lowest blood pH, compared with SARA-negative herds, were observed. The lowering of ruminal fluid pH increased the blood IGF-I and NEFA concentrations and the rectal temperature and decreased the blood pH. These measures are indicators of the physiological changes that occur as part of the pathogenesis of the condition and may be helpful for the diagnosis of the SARA syndrome when serial measurements are conducted.

Memetic algorithms for ligand expulsion from protein cavities.

Ligand diffusion through a protein interior is a fundamental process governing biological signaling and enzymatic catalysis. A complex topology of channels in proteins leads often to difficulties in modeling ligand escape pathways by classical molecular dynamics simulations. In this paper, two novel memetic methods for searching the exit paths and cavity space exploration are proposed: Memory Enhanced Random Acceleration (MERA) Molecular Dynamics (MD) and Immune Algorithm (IA). In MERA, a pheromone concept is introduced to optimize an expulsion force. In IA, hybrid learning protocols are exploited to predict ligand exit paths. They are tested on three protein channels with increasing complexity: M2 muscarinic G-protein-coupled receptor, enzyme nitrile hydratase, and heme-protein cytochrome P450cam. In these cases, the memetic methods outperform simulated annealing and random acceleration molecular dynamics. The proposed algorithms are general and appropriate in all problems where an accelerated transport of an object through a network of channels is studied.

Communication: Entropic measure to prevent energy over-minimization in molecular dynamics simulations.

This work examines the impact of energy over-minimization on an ensemble of biological molecules subjected to the potential energy minimization procedure in vacuum. In the studied structures, long potential energy minimization stage leads to an increase of the main- and side-chain entropies in proteins. We show that such over-minimization may diverge the protein structures from the near-native attraction basin which possesses a minimum of free energy. We propose a measure based on the Pareto front of total entropy for quality assessment of minimized protein conformation. This measure may help in selection of adequate number of energy minimization steps in protein modelling and, thus, in preservation of the near-native protein conformation.

Study on the zona pellucida 4 (ZP4) gene sequence and its expression in the ovaries of patients with polycystic ovary syndrome.

BACKGROUND: Polycystic ovary syndrome (PCOS) is a common endocrine disorder of unknown pathology, involving reproductive and metabolic abnormalities. Oocyte-specific genes are a group of genes expressed exclusively in ovarian tissue; therefore, they can play an important role in ovarian pathologies such as PCOS. The zona pellucida 4 (ZP4) gene encodes glycoprotein which is a part of the extracellular matrix of oocyte. MATERIALS AND METHODS: We analyzed 87 patients with PCOS, which were divided into four groups depending on their phenotype. In each patient, we performed profound clinical and biochemical analysis, including the measurement of serum androgens. The ovarian tissue samples were used to perform a real-time polymerase chain reaction and immunohistochemical staining using anti-ZP4 monoclonal antibodies. The ZP4 gene was sequenced from peripheral lymphocytes. RESULTS: The expression of ZP4 was present in early antral follicles and was stronger in mature follicles. The subgroup of patients with eumenorrhea and without hyperandrogenism presented the highest expression of ZP4 in ovarian tissue. In one case, we found a mutation of the ZP4 gene. No correlations were found between the ZP4 expression level and biochemical or clinical indices. CONCLUSIONS: Data from this and animal studies suggest a possible relationship between androgens and ZP4 expression. ZP4 expression is highest among patients with PCOS and a regular cycle, and this is a consequence of the presence of mature follicles in this group. In some patients with PCOS and infertility, ZP4 mutation can be found.

Graded or random - Effect of pore distribution in 3D titanium scaffolds on corrosion performance and response of hMSCs.

Researchers agree that the ideal scaffold for tissue engineering should possess a 3D and highly porous structure, biocompatibility to encourage cell/tissue growth, suitable surface chemistry for cell attachment and differentiation, and mechanical properties that match those of the surrounding tissues. However, there is no consensus on the optimal pore distribution. In this study, we investigated the effect of pore distribution on corrosion resistance and performance of human mesenchymal stem cells (hMSC) using titanium scaffolds fabricated by laser beam powder bed fusion (PBF-LB). We designed two scaffold architectures with the same porosities (i.e., 75 %) but different distribution of pores of three sizes (200, 500, and 700 µm). The pores were either grouped in three zones (graded, GRAD) or distributed randomly (random, RAND). Microfocus X-ray computed tomography revealed that the chemically polished scaffolds had the porosity of 69 ± 4 % (GRAD) and 71 ± 4 % (RAND), and that the GRAD architecture had the higher surface area (1580 ± 101 vs 991 ± 62 mm2) and the thinner struts (221 ± 37 vs 286 ± 14 µm). The electrochemical measurements demonstrated that the apparent corrosion rate of chemically polished GRAD scaffold decreased with the immersion time extension, while that for polished RAND was increased. The RAND architecture outperformed the GRAD one with respect to hMSC proliferation (over two times higher although the GRAD scaffolds had 85 % higher initial cell retention) and migration from a monolayer. Our findings demonstrate that the pore distribution affects the biological properties of the titanium scaffolds for bone tissue engineering.

Prophylactic central neck dissection for papillary thyroid cancer.

BACKGROUND: Prophylactic central neck dissection (CND) for papillary thyroid cancer (PTC) remains controversial. The aim of this study was to examine whether prophylactic CND for PTC affected long-term survival and locoregional control. METHODS: This was a retrospective cohort study of patients who underwent total thyroidectomy (TT) with bilateral prophylactic CND. They were compared with patients who had TT without CND. Personalized adjuvant radioiodine treatment was used in both groups. Primary outcomes were overall and disease-specific survival, and locoregional control. Secondary outcomes were number of patients with negative serum thyroglobulin levels, and morbidity. RESULTS: Of 640 patients with PTC included in this study, 282 (treated in 1993-1997) had TT without CND and 358 (treated in 1998-2002) underwent TT with CND. The 10-year disease-specific survival rate for patients who had TT without CND was 92·5 per cent compared with 98·0 per cent in patients with CND (P = 0·034), and the locoregional control rate was 87·6 and 94·5 per cent respectively (P = 0·003). In multivariable analysis, extrathyroidal extension was an independent predictive factor for locoregional recurrence (odds ratio 12·47, 95 per cent confidence interval 6·74 to 23·06; P < 0·001), whereas CND was an independent predictive factor for improved locoregional control at 10 years after surgery (odds ratio 0·21, 0·11 to 0·41; P < 0·001). No differences were seen in the rates of permanent hypoparathyroidism or recurrent laryngeal nerve injury between the groups. CONCLUSION: Bilateral prophylactic CND for staging of the neck in PTC, followed by personalized adjuvant radioiodine treatment, improved both 10-year disease-specific survival and locoregional control, without increasing the risk of permanent morbidity. REGISTRATION NUMBER: NCT01510002 (http://www.clinicaltrials.gov).

Dietary energy density in the dry period on the metabolic status of lactating cows.

The aim of the study was to investigate the effect of different energy concentrations in the isonitrogenous diet fed during the dry period on postpartum health, fertility and blood variables. Forty Holstein multiparous cows were dried 56 days before the expected day of calving and assigned to group (M) with moderate energy concentrations of 0.69 UFL/kg DM or to the low-energy group (L) with energy density of 0.61 UFL/kg DM. From the 7d before the expected day of calving until the 21d of lactation, all the cows were fed the same fresh transition diet (0.82 UFL/kg DM). From the 22d to the 90d of lactation, all the cows received the same highest energy-density lactation diet (0.90 UFL/kg DM). During the dry period the decline of BCS in groups M and L were 0.07 and 0.12 units respectively. The average decrease of BCS from calving to 56 d of lactation were the same in both experimental groups (0.21 BCS). The first-service conception rate tended to be higher in the M group. Insulin-like growth factor-1, glucose, beta-hydroxybutyric acid, non-esterified fatty acid, thyroxine serum concentrations prepartum and 3 and 5d postpartum were not significantly affected by the treatment in the dry period.

Randomized clinical trial of bilateral subtotal thyroidectomy versus total thyroidectomy for Graves' disease with a 5-year follow-up.

BACKGROUND: The extent of thyroid resection in Graves' disease remains controversial. The aim of this study was to evaluate long-term results of bilateral subtotal thyroidectomy (BST) compared with total thyroidectomy (TT) in patients with Graves' disease and mild active ophthalmopathy. METHODS: Participants were assigned randomly to BST or TT, and followed for 5 years after surgery. The primary endpoints of the study were the prevalence of recurrent hyperthyroidism and changes in Graves' ophthalmopathy. Secondary endpoints were postoperative transient and permanent paresis of the recurrent laryngeal nerve, and postoperative hypocalcaemia and hypoparathyroidism. RESULTS: Two hundred patients were included, of whom 191 (BST 95, TT 96) completed the 5-year follow-up. Recurrent hyperthyroidism occurred in nine patients after BST and in none after TT (P = 0·002). Progression of Graves' ophthalmopathy was observed in nine patients after BST compared with seven following TT (P = 0·586). Transient hypoparathyroidism occurred in 13 and 24 patients respectively (P = 0·047). Permanent hypoparathyroidism was diagnosed in no patient after BST and in one after TT (P = 0·318). No differences were noted in transient or permanent recurrent laryngeal nerve injury. CONCLUSION: TT for Graves' disease prevented recurrent hyperthyroidism but did not prevent the progression of ophthalmopathy compared with BST.

The impact of cow nutrition in the dry period on colostrum quality and immune status of calves.

The objective of the study was to evaluate the effect of energy contents in a dry period diets in "7+1" feeding strategy of dry cow including 7 weeks of the dry period (far-off) and 1 week of a close-up period, on colostrum quality and the immune status of calves. Forty Holstein multiparous cows were dried at 56d before the expected date of calving and were assigned to the higher energy diet group (HE; 0.69 UFL/kg DM, NDF 52% DM), or the lower energy diet group (LE; 0.61 UFL/1kg DM, NDF 56% DM). From -7d to the expected calving date up to 21 d of lactation, all cows were fed the same fresh, lactation diet. Samples of colostrum were collected within 2h after parturition and its density, dry matter content and concentrations of immunoglobulins, fat, protein, lactose, urea and somatic cell counts were measured. Calves were weighed 2 h after calving and on the 21d of life. On 3d and 21d of calves' life blood samples were collected and the concentration of IgG, IgA, IgM, insulin-like growth factor 1 (IGF-1) as well as total protein and albumin concentrations were determined. Treatments had no significant effect on composition of colostrum and serum immunoglobulins and IGF-1 concentration. In both groups the weight of calves at birth was similar, still daily body weight gain during 21 day of rearing period in HE group was higher than in the LE group (P=0.05).

Association of the IGF-I promoter P1 polymorphism with risk of cervical cancer.

Human papillomaviruses (HPV16, HPV18, HPV31, HPV33) are etiological agents in the development of cervical cancer. HPVs infect epithelial cells and depend on epithelial differentiation for the completion of their life cycle. Insulin-like growth factor I (IGF-I) is a potent mitogen involved in the regulation of cell proliferation and apoptosis of many cell types including normal and transformed epithelial cells. Deregulation of IGF-I expression and action is linked to diverse pathologies including cancer. A polymorphism in the P1 promoter region of the IGF-I gene may directly influence its expression. Using the PCR-SSCP method and sequencing of DNA, we identified a single nucleotide polymorphism (SNP) at -383(C>T) position of promoter P1 of the IGF-I in 16% of the study HPV-positive women with precancerous and cancerous lesions. In vitro, we observed that the SNP at-383(C>T) site significantly increased the reporter gene expresion in the HepG2 cell line, but not in the HeLa cell line relative to the wild type promoter. It suggests that the studied SNP can change expression of the IGF-I gene in distinct ways in different types of tissues. Deregulation of expression of the IGF-I gene can affect normal epithelium development and in case of HPV infection can potentially disrupt the virus life cycle and stimulate its passage into the oncogenic life cycle or persistent viral infections. Therefore, we propose that SNP C>T at the -383 position of P1 promoter may be one of the helpful prognostic markers in the diagnosis of cervical cancer development of women with persistent infection in the ectocervical epithelium. We have not found any association between the polymorphism CA repeats in the promoter P1 region of the IGF-I gene and suceptibility to HPV infection and cervical cancer development. The (CA)19 allele was the most common in the study of this group of women.

Effects of different starch sources on metabolic profile, production and fertility parameters in dairy cows.

The objective of the study was to determine the effect of replacing triticale (high rumen degradable starch) with maize grain (low rumen degradable starch) during the transition period and the first 120 days of lactation on metabolic and hormonal profile indices, milk production and fertility performance in cows. Forty-eight Holstein-Friesian dairy cows were divided into 4 groups: TT (2.5 kg triticale grain/cow per day supplemented from 14 days prepartum to day 120 postpartum), TM (2.5 kg triticale grain/cow per day supplemented from day 14 before parturition to calving, and then 2.5 kg maize grain to 120 days of lactation), MT (2.5 kg maize grain/cow per day supplemented from day 14 before parturition to calving, and then 2.5 kg triticale grain to 120 days of lactation), MM (2.5 kg maize grain/cow per day supplemented from 14 days prepartum to day 120 postpartum). Blood samples were collected 3 weeks and 1 week before calving and on days 14, 56 and 70 of lactation, and they were analyzed in terms of concentrations of glucose, insulin, leptin, insulin-like growth factor I, nonesterified fatty acids, triglycerides, cholesterol, blood urea nitrogen and activities of aspartate aminotransferase and gamma glutamyl transpeptidase. Milk samples were collected twice a day at weekly intervals and analyzed for fat, protein and lactose. Milk yield and individual dry mater intake were recorded at weekly intervals. Body condition was estimated 3 weeks before calving, on parturition day and on days 14, 56 and 120 of lactation. Replacing triticale grain with maize grain in the transition period and during lactation positively affected fertility of lactating cows. An increased first service conception rate and shortening of the days open period was observed in MM and TM groups in comparison to those found in group MT (P < or = 0.05). The lowest number of services per conception was recorded in groups MM and TM (P < or = 0.05). Although the impact of milk production and the most of the blood indices were not significantly affected by this treatment, the results of the study suggest that maize grain in the transition period and lactation might be a more effective energy source for dairy cows than triticale grain.

Doppler guided haemorrhoidal arterial ligation with recto-anal-repair (RAR) for the treatment of advanced haemorrhoidal disease.

OBJECTIVE: A modification of Doppler guided haemorrhoidal artery ligation (DGHAL) to include the addition of recto-anal repair is reported. Preliminary results of function and safety of third and fourth degree haemorrhoidals are given. METHOD: Thirty patients underwent DGHAL combined with recto-anal-repair (RAR). Each had rectal examination, anorectal manometry and Quality of Life assessment before and 3 months after the procedure. RESULTS: Twenty-nine patients were included in the final analysis. There were three (10.34%) patients of intra-operative and one (3.45%) of postoperative bleeding. Three months after RAR (17.24%) patients with minor residual mucosal prolapse were detected, three (10.34%) patients reported residual symptoms. There was no case of recurrent bleeding. Anal manometry at 3 months after RAR was significantly lower than before the procedure (P < 0.05). One (3.45%) patient reported occasional soiling 3 months after RAR. CONCLUSION: Recto-anal-repair is safe in treating third and fourth degree haemorrhoids with no major complications and low rate of residual disease.

Effect of oat hay provision method on growth performance, rumen fermentation and blood metabolites of dairy calves during preweaning and postweaning periods.

The comparison of the effects of all forage offering methods would be particularly useful information in modeling growth performance and rumen fermentation of dairy calves. Therefore, this study attempted to evaluate the effects of methods of oat hay provision on growth performance, rumen fermentation and biochemical blood indices of dairy calves during preweaning and postweaning periods. At birth, 40 female Polish Holstein-Friesian calves (3 days of age; 39.6 ± 0.39 kg BW) were randomly assigned to four treatment groups differing in the access to chopped oat hay: CON (control, starter without oat hay), OH (starter feed containing 10% DM basis oat hay), OH-FC (starter feed containing 10% DM basis oat hay and oat hay fed as free-choice provision in different buckets) and FC (starter feed and oat hay fed as free-choice provision in different buckets). The calves were weaned on day 56, and then the study continued until day 84. Intakes of starter feed and oat hay were recorded daily, whereas BW and hip height (HH) on day 3 and then every 14 days. Samples of blood were collected on the initiation of experiment and then every 14 days, and rumen contents on day 28, 56 and 84. No treatment effects were found for starter, starch, CP, total DM intake, average daily gain, feeding efficiency, change in HH, ruminal fluid pH, concentrations of ruminal propionate and NH3-N, concentrations of urea nitrogen and non-esterified fatty acids in the blood. There were differences between treatments in terms of ruminal total volatile fatty acids and molar concentrations of acetate, butyrate and acetate to propionate ratio; highest in OH and OH-FC groups, especially during the postweaning period. On the other hand, lower concentrations of iso-valerate were found in OH and OH-FC groups on day 56 and 84. The concentrations of IGF-I throughout the experiment and ß-hydroxybutyrate during the postweaning period in the blood were influenced by treatment, with the greatest values observed in OH and OH-FC calves. Results of this study indicate that starter feed containing chopped oat hay improves rumen fermentation parameters, which might allow successful transition from preruminant to mature ruminant state. Also, providing chopped oat hay with pelleted starter feed seems to be a better method than free-choice supplementation.

Intracellular nanomanipulation by a photonic-force microscope with real-time acquisition of a 3D stiffness matrix.

A traditional photonic-force microscope (PFM) results in huge sets of data, which requires tedious numerical analysis. In this paper, we propose instead an analog signal processor to attain real-time capabilities while retaining the richness of the traditional PFM data. Our system is devoted to intracellular measurements and is fully interactive through the use of a haptic joystick. Using our specialized analog hardware along with a dedicated algorithm, we can extract the full 3D stiffness matrix of the optical trap in real time, including the off-diagonal cross-terms. Our system is also capable of simultaneously recording data for subsequent offline analysis. This allows us to check that a good correlation exists between the classical analysis of stiffness and our real-time measurements. We monitor the PFM beads using an optical microscope. The force-feedback mechanism of the haptic joystick helps us in interactively guiding the bead inside living cells and collecting information from its (possibly anisotropic) environment. The instantaneous stiffness measurements are also displayed in real time on a graphical user interface. The whole system has been built and is operational; here we present early results that confirm the consistency of the real-time measurements with offline computations.