Comparing responses of dairy cows to short-term and long-term heat stress in climate-controlled chambers.

Heat stress (HS) in dairy cows can be classified into short-term heat stress (STHS) and long-term heat stress (LTHS) according to the number of consecutive days in HS. The comparative study of these 2 types of HS is limited in terms of their effects on the production and energy metabolism of cows. In this study, 4 lactating Holstein cows (102.5 ± 12 days in milk, 605 ± 22 kg of body weight, second parity) fitted with rumen fistulae were randomly assigned to 1 of 2 groups in a 2 × 2 crossover design and allocated to 1 of 2 climate-controlled chambers. This study contained 2 periods, each with a control phase and a HS phase. There was a recovery phase between 2 periods. The HS phase comprised either STHS (3 d) or LTHS (7 d) treatments. Data collected from the 3 d of STHS and the last 3 d of LTHS were compared. The chambers were set at thermal neutral conditions (20°C, 50% humidity) during the control and recovery phases or cyclical HS conditions (26-38°C, 50% humidity) during the HS phase. Compared with STHS, LTHS decreased milk yield by 17.2% and dry matter intake by 12.6%, indicating that LTHS caused a more severe decline in milk production and feed intake. In addition, LTHS decreased milk protein concentration by 6.8% and milk protein yield by 22.4%. In comparison with STHS, LTHS decreased rumen liquor volatile fatty acid (29.7%), blood glucose (11.6%), and nonesterified fatty acid (13.6%) concentrations, but increased milk urea nitrogen by 15.1%, blood urea nitrogen by 8.6%, and creatine concentrations by 15.4%. Our results suggest that although reduced feed intake may be mainly responsible for reduced milk production during STHS, impaired rumen metabolism and suppressed mobilization of adipose tissue could be the main reasons for further reduction in milk yield during LTHS.

Long non-coding RNA expression profile in permanent atrial fibrillation patients with rheumatic heart disease.

OBJECTIVE: Atrial fibrillation (AF) is the most common type of arrhythmia, especially in rheumatic heart disease (RHD) patients. The differences in structural remodeling and electrical remodeling between the left and right atrium associated with AF in RHD patients are well known, and alterations in the expression profiles of long noncoding RNAs (lncRNAs) in the left atrium have also been investigated. However, the role of lncRNAs in the right atrium (RA) remains largely unknown. PATIENTS AND METHODS: We identified differentially expressed lncRNAs in RA tissues of RHD patients with AF or a normal sinus rhythm (NSR) using microarray analysis. Then, we performed gene ontology (GO) and KEGG pathway analyses for functional annotation of the deregulated lncRNAs. Finally, we constructed a lncRNA-mRNA co-expression network. RESULTS: Of the 22,829 human non-coding RNAs analyzed, a total of 1,909 long non-coding RNAs were detected. A total of 182 lncRNAs (117 downregulated and 65 upregulated) were shown to be differentially expressed (fold-change > 1.5) in AF patients compared with NSR patients. Many lncRNAs might be partially involved in an AF-related pathway. CONCLUSIONS: AF dysregulates the expression of lncRNAs in the RA of RHD patients. These findings may be useful for exploring potential therapeutic treatments for AF in RHD patients.

[Screening for precursors of colorectal cancer].

Screening has been proven to be effective for the control of colorectal cancer (CRC). The target of CRC screening is shifting from CRC to colorectal neoplasia (CN), the precursors of CRC. Based on the the latest national guideline, the Consensus of Screening for CRC and CN, and the recent research of precursors both at home and abroad. This paper summarizes the progress in the research of risk factors, risk prediction model, screening strategy optimization, colonoscopy quality control, sessile serrated adenoma identification and follow up as well as the recognition of precursors.

ATP-sensitive potassium currents in rat primary afferent neurons: biophysical, pharmacological properties, and alterations by painful nerve injury.

ATP-sensitive potassium (K(ATP)) channels may be linked to mechanisms of pain after nerve injury, but remain under-investigated in primary afferents so far. We therefore characterized these channels in dorsal root ganglion (DRG) neurons, and tested whether they contribute to hyperalgesia after spinal nerve ligation (SNL). We compared K(ATP) channel properties between DRG somata classified by diameter into small or large, and by injury status into neurons from rats that either did or did not become hyperalgesic after SNL, or neurons from control animals. In cell-attached patches, we recorded basal K(ATP) channel opening in all neuronal subpopulations. However, higher open probabilities and longer open times were observed in large compared to small neurons. Following SNL, this channel activity was suppressed only in large neurons from hyperalgesic rats, but not from animals that did not develop hyperalgesia. In contrast, no alterations of channel activity developed in small neurons after axotomy. On the other hand, cell-free recordings showed similar ATP sensitivity, inward rectification and unitary conductance (70-80 pS) between neurons classified by size or injury status. Likewise, pharmacological sensitivity to the K(ATP) channel opener diazoxide, and to the selective blockers glibenclamide and tolbutamide, did not differ between groups. In large neurons, selective inhibition of whole-cell ATP-sensitive potassium channel current (I(K(ATP))) by glibenclamide depolarized resting membrane potential (RMP). The contribution of this current to RMP was also attenuated after painful axotomy. Using specific antibodies, we identified SUR1, SUR2, and Kir6.2 but not Kir6.1 subunits in DRGs. These findings indicate that functional K(ATP) channels are present in normal DRG neurons, wherein they regulate RMP. Alterations of these channels may be involved in the pathogenesis of neuropathic pain following peripheral nerve injury. Their biophysical and pharmacological properties are preserved even after axotomy, suggesting that K(ATP) channels in primary afferents remain available for therapeutic targeting against established neuropathic pain.

Ontogeny of circadian and light regulation of melatonin release in Xenopus laevis embryos.

The retinal photoreceptors of Xenopus laevis contain a circadian clock that controls the synthesis and release of melatonin, resulting in high levels during the night and low levels during the day. Light is also an important regulator of melatonin synthesis and acts directly to acutely suppress melatonin synthesis during the day and indirectly to entrain the circadian clock. We examined the development of circadian and light regulation of melatonin release in Xenopus retinas and pineal glands. Pineal glands are capable of making measurable melatonin in culture soon after they evaginate from the diencephalon at stage 26. In cyclic light, the melatonin rhythms are robust, with higher overall levels and greater amplitudes than in constant darkness. However, the rhythm of melatonin release damps strongly and quickly toward baseline in constant darkness. Similar results are observed in older (stage 47) embryos, indicating that cyclic light has a positive effect on melatonin synthesis in this tissue. Optic vesicles dissected at stage 26 do not release melatonin in culture until the second or third day. It is weakly rhythmic in cyclic light, but in constant dark it is released at constitutively high levels throughout the day. By stage 41, the eyes release melatonin rhythmically in both cyclic light and constant darkness with similar amplitude. Our results show that Xenopus embryos develop a functional, photoresponsive circadian clock in the eye within the first few days of life and that rhythmic melatonin release from the pineal gland at comparable stages is highly dependent on a light-dark cycle.

Linkage of a nucleolin-related protein and casein kinase II with the detergent-stable photoreceptor cytoskeleton.

Vertebrate photoreceptors are highly polarized sensory neurons with a complex microtubule and actin-based cytoskeletal organization. In the present study, we have used a detergent-extracted cytokeleton preparation from bovine photoreceptors to test the hypothesis that protein kinases and their substrates co-purify with the photoreceptor cytoskeleton. We incubated the cytoskeletal preparation in the presence of [gamma-32P]ATP. Following SDS-PAGE and autoradiography, we found two principal phosphoproteins with apparent molecular weights of 55 kDa (pp55) and 112 kDa (pp112). We have additionally identified the kinase responsible for phosphorylation of pp112 (and possibly pp55) as a casein kinase II-like enzyme. pp55 was identified as beta-tubulin based on Western blotting and its position on two-dimensional gels. Microsequencing revealed that 16 of the first 17 amino acids of pp112 were identical to human nucleolin, a nuclear protein. Western blotting, mobility in SDS PAGE and in two-dimensional gels, predominant localization within the nucleus, and phosphorylation by a casein kinase II all support the conclusion that pp112 is a nucleolin-related protein. Immunocytochemistry revealed a significant extranuclear pool of nucleolin-immunoreactivity within the cell bodies of photoreceptors. These findings suggest an important extranuclear role for nucleolin or a related protein in photoreceptors.

Oxidized low-density lipoprotein decreases the induced nitric oxide synthesis in rat mesangial cells.

Recently, the close relation between oxidized low density lipoprotein (Ox-LDL) and the progression of glomerular injury has been demonstrated. The nitric oxide (NO) pathway in glomerular mesangial cells may be a potential target for the adverse effects of Ox-LDL in the development of glomerular injury. In this study, we treated cultured rat mesangial cells (RMC) with Fe(2+)-oxidized LDL and then stimulated the cells with lipopolysacharride (LPS, 10 micrograms ml-1). The LPS-induced NO production, assessed by NO2-concentrations in cultured supernatants, decreased from 7.83 nmol per 10(6) cells in control to 4.00 nmol per 10(6) cells and 1.67 nmol per 10(6) cells in RMC preincubated with Ox-LDL at 20 micrograms ml-1 and 40 micrograms ml-1, respectively (P < 0.01). Native LDL had no significant effects on LPS-induced NO production. Using the reverse transcription-polymerase chain reaction (RT-PCR) technique, we could not detect significant alteration of inducible NO synthase (iNOS) mRNA levels in RMC preincubated with Ox-LDL. Our results suggest that Ox-LDL decreases induced NO production in RMC, which may contribute to the adverse effects of Ox-LDL in progressive glomerular injury. The mechanisms of this decrease may not involve changes of iNOS genic transcription.

Ocular retardation mouse caused by Chx10 homeobox null allele: impaired retinal progenitor proliferation and bipolar cell differentiation.

Ocular retardation (or) is a murine eye mutation causing microphthalmia, a thin hypocellular retina and optic nerve aplasia. Here we show that mice carrying the OrJ allele have a premature stop codon in the homeobox of the Chx10 gene, a gene expressed at high levels in uncommitted retinal progenitor cells and mature bipolar cells. No CHX10 protein was detectable in the retinal neuroepithelium of orJ homozygotes. The loss of CHX10 leads both to reduced proliferation of retinal progenitors and to a specific absence of differentiated bipolar cells. Other major retinal cell types were present and correctly positioned in the mutant retina, although rod outer segments were short and retinal lamination was incomplete. These results indicate that Chx10 is an essential component in the network of genes required for the development of the mammalian eye, with profound effects on retinal progenitor proliferation and bipolar cell specification or differentiation. off

[Changes in serum folic acid and vitamin B12 levels in liver cirrhosis and its clinical significance].

The levels of serum folic acid and vitamin B12 were determined in 40 cases of liver cirrhosis with radioimmunoassay. It was shown that in 87.5% of the patients folic acid level was lower than that of a control group and in 67.5% serum vitamin B12 level was higher than that of the control group (P less than 0.05). The correlation between liver cirrhosis and dysbolism of folic acid and vitamin B12 and the megaloblastic changes and clinical significance were discussed.

Production and characterization of antibody against diacetoxyscirpenol.

Antibodies against diacetoxyscirpenol (DAS) were obtained from rabbits after immunizing them with hemisuccinate or hemiglutarate derivatives of DAS conjugated to bovine serum albumin (BSA). DAS-hemiglutarate-BSA was found to be a much better immunogen than DAS-hemisuccinate-BSA. Competitive radioimmunoassay revealed that the antisera obtained from rabbits after immunization with DAS-hemiglutarate-BSA showed high specificity toward DAS. The concentrations causing 50% displacement of radioactive DAS by unlabeled DAS, 4-monoacetoxyscirpenol (MAS), and 15-MAS were found to be 1.5, 130, and 300 ng per assay, respectively. Thus, the cross-reactivities for 4-MAS and 15-MAS are ca. 87 and 300 times weaker than that of DAS. Practically no cross-reaction (less than 5% displacement) was observed when deoxynivalenol, T-2 toxin, deoxyverrucarol, and scirpentriol were tested at a concentration of 2,000 ng/ml.

Screening uremic 'toxins' using bromsulfophthalein clearance by the isolated perfused rat liver.

The kinetics of bromosulfophthalein clearance from plasma, secretion into bile, and, by difference, storage in hepatic cells, were determined, using normal livers and blood from normal rats in the isolated perfused rat liver system. Two suspected uremic 'toxins', urea and guanidinosuccinic acid, were then administered in order to determine if either of these substances would independently alter the kinetics under study. Results indicated that these two compounds have little or no effect on liver function on an acute basis. However, because of the excellent reproducibility obtained, it is believed that this method can be of significant use in the further screening of substances from uremic patients which may be interfering seriously with liver functions.