Delta-frequency stimulation of cerebellar projections can compensate for schizophrenia-related medial frontal dysfunction.

Schizophrenia involves abnormalities in the medial frontal cortex that lead to cognitive deficits. Here we investigate a novel strategy to normalize medial frontal brain activity by stimulating cerebellar projections. We used an interval timing task to study elementary cognitive processing that requires both frontal and cerebellar networks that are disrupted in patients with schizophrenia. We report three novel findings. First, patients with schizophrenia had dysfunctional delta rhythms between 1-4 Hz in the medial frontal cortex. We explored cerebellar-frontal interactions in animal models and found that both frontal and cerebellar neurons were modulated during interval timing and had delta-frequency interactions. Finally, delta-frequency optogenetic stimulation of thalamic synaptic terminals of lateral cerebellar projection neurons rescued timing performance as well as medial frontal activity in a rodent model of schizophrenia-related frontal dysfunction. These data provide insight into how the cerebellum influences medial frontal networks and the role of the cerebellum in cognitive processing.

Epigenetic suppression of potassium-chloride co-transporter 2 expression in inflammatory pain induced by complete Freund's adjuvant (CFA).

BACKGROUND: Multiple mechanisms contribute to the stimulus-evoked pain hypersensitivity that may be experienced after peripheral inflammation. Persistent pathological stimuli in many pain conditions affect the expression of certain genes through epigenetic alternations. The main purpose of our study was to investigate the role of epigenetic modification on potassium-chloride co-transporter 2 (KCC2) gene expression in the persistence of inflammatory pain. METHODS: Persistent inflammatory pain was induced through the injection of complete Freund's adjuvant (CFA) in the left hind paw of rats. Acetyl-histone H3 and H4 level was determined by chromatin immunoprecipitation in the spinal dorsal horn. Pain behaviour and inhibitory synaptic function of spinal cord were determined before and after CFA injection. KCC2 expression was determined by real time RT-PCR and Western blot. Intrathecal KCC2 siRNA (2 µg per 10 µL per rat) or HDAC inhibitor (10 µg per 10 µL per rat) was injected once daily for 3 days before CFA injection. RESULTS: Persistent inflammatory pain epigenetically suppressed KCC2 expression through histone deacetylase (HDAC)-mediated histone hypoacetylation, resulting in decreased inhibitory signalling efficacy. KCC2 knock-down caused by intrathecal administration of KCC2 siRNA in naïve rats reduced KCC2 expression in the spinal cord, leading to sensitized pain behaviours and impaired inhibitory synaptic transmission in their spinal cords. Moreover, intrathecal HDAC inhibitor injection in CFA rats increased KCC2 expression, partially restoring the spinal inhibitory synaptic transmission and relieving the sensitized pain behaviour. CONCLUSION: These findings suggest that the transcription of spinal KCC2 is regulated by histone acetylation epigenetically following CFA. SIGNIFICANCE: Persistent pain suppresses KCC2 expression through HDAC-mediated histone hypoacetylation and consequently impairs the inhibitory function of inhibitory interneurons. Drugs such as HDAC inhibitors that suppress the influences of persistent pain on the expression of KCC2 may serve as a novel analgesic.

Curcumin attenuates airway hyperreactivity induced by ischemia-reperfusion of the pancreas in rats.

OBJECTIVES: Ischemia-reperfusion (I/R) of the rat pancreas induces acute pancreatitis with a systemic inflammatory response. Activated inflammatory cells are sequestered in the lung, and the consequent respiratory burst may increase airway reactivity. In this study, we characterized the effect of the antioxidant curcumin on airway hyperreactivity induced by pancreatic I/R. METHODS: Ischemia of the pancreas was induced by clamping the gastroduodenal and the splenic artery for 2 hours followed by reperfusion for 6 hours. The pulmonary function data of Penh, a measurement of airway resistance, were used to show the airway responses to a methacholine challenge. The blood concentration of oxygen radicals, nitric oxide, and tumor necrosis factor-alpha (TNFalpha) were measured after pancreatic I/R. mRNA expressions of inducible nitric oxide synthase (iNOS) and TNFalpha in lung tissues were measured after pancreatic I/R. Pretreatment with curcumin (20 mg/kg) was administered by intraperitoneal injection 2 hours before pancreatic I/R. RESULTS: The protocol resulted in significant elevations of the blood concentrations of amylase, hydroxyl radical, nitric oxide, TNFalpha, and white cells among the I/R group. iNOS and TNFalpha mRNA expressions also significantly increased in lung tissues. Pulmonary function data showed that pancreatic I/R induced significant increases in responses to methacholine challenge: Penh increased significantly in the I/R group when compared with the sham group. Pretreatment with curcumin significantly attenuated the inflammatory, oxidative, and nitrosative responses and lung tissue iNOS and TNFalpha expressions. Curcumin also attenuated airway reactivity to methacholine challenge. CONCLUSIONS: I/R of the pancreas induced systemic inflammatory responses with respiratory burst, nitrosative stress, and hyperresponses in the airways. Curcumin, which has antioxidant and anti-inflammatory effects, significantly attenuated the inflammatory responses and airway hyperreactivity induced by pancreatic I/R.

The effect of glutamine-supplemented total parenteral nutrition on nitrogen economy depends on severity of diseases in surgical patients.

BACKGROUND: Gln is an important substrate for enterocyte and rapid proliferation cells. Studies have shown that parenteral supplementation of Gln maintains the intracellular Gln pool, improves nitrogen balance and shortens hospital stay. However, some studies showed Gln-supplemented TPN had no effect on restoring the Gln pool in critically ill patients. OBJECTIVE: To evaluate the effect of glutamine (Gln) dipeptide supplementation of total parenteral nutrition (TPN) on postoperative nitrogen balance and immune response of patients undergoing surgery. METHODS: This study is a prospective, randomized double-blind clinical trial. APACHE II score and TISS were used to evaluate the patients after admission. Forty-eight patients with major abdominal surgery were allocated to two groups to receive isonitrogenous (0.228 g nitrogen/kg/day) and isoenergetic (30 kcal/kg/day) TPN for 6 days. Two groups (Conv and Ala-Gln) were further divided to high (APACHE>or=6) and low (APACHE <6) groups. Control group (Conv) received 1.5 g amino acids/kg/day, whereas the Ala-Gln group received 0.972 g amino acids/kg/day and 0.417 g of L-alanyl-L-glutamine (Ala-Gln)/kg/day. Blood samples were collected on day 1 and day 6 after surgery for plasma amino acid and CD4, CD8 cell and T lymphocyte analysis. Cumulative nitrogen balance were also measured on day 2, 3, 4, 5 postoperatively. RESULTS: Although there was a tendency to have better cumulative nitrogen balance on the postoperative days in the Ala-Gln group, no significant difference was observed between two groups. However, a better significant cumulative nitrogen balance was observed on the 2nd, 3rd and 5th postoperative day in the Ala-Gln group than in the Conv group in patients with APACHE II <6, whereas no significant difference was noted in patients with APACHE II >or= 6. No difference in urine 3-methylhistidine excretion were observed between the 2 groups. Patients in the Ala-Gln group had significant higher T lymphocyte and CD4 cells than did those in the Conv group. CONCLUSION: TPN supplemented with Gln dipeptide had beneficial effect on enhancing the immune response. However, the effect of Ala-Gln administration on improving nitrogen economy was only observed in patients with low APACHE II scores. These results may indicate that Gln required for reversing the catabolic condition may depend on the characteristics and severity of the diseases.

Influence of sorghum grain processing on performance and nutrient digestibilities in dairy cows fed varying concentrations of fat.

Forty Holstein cows averaging 85 +/- 50 d in milk were assigned to five dietary treatments for 56 d to determine the effects of ruminal starch degradability and supplemental fat on milk yield and composition and nutrient digestibilities. Treatments were 1) dry-rolled sorghum, no added fat; 2) dry-rolled sorghum plus 2.5% prilled fatty acids; 3) steam-flaked sorghum, no added fat; 4) steam-flaked sorghum plus 2.5% prilled fatty acids; and 5) steam-flaked sorghum plus 5% prilled fatty acids. Fat supplementation at 2.5 and 5% increased the content of fatty acids in diets from 3.0 to 5.4 and 7.7%, respectively. Milk yield was increased 2.0 kg/d when steam-flaked sorghum plus 0 or 2.5% added fat was fed and was 2.4 kg/d higher when prilled fatty acids were fed at 2.5 versus 0%. Cows fed supplemental fat at 5% did not increase milk yield over that of cows fed no added fat. For cows fed 2.5% fat, steam-flaking decreased dry matter intake 12% and increased efficiency of conversion of feed to milk 11%. Milk composition was unaffected by treatments, except that lactose was depressed by fat supplementation. Milk protein yield and efficiency of conversion of dietary protein to milk protein were increased when steam-flaked sorghum was fed. Starch digestibilities were increased from 92.6 to 98.2% when sorghum was steam-flaked compared with dry-rolled and fat supplementation tended to decrease digestibility of neutral detergent fiber. Steam-flaking tended to improve, and the addition of 2.5% fat did improve, the lactational performances of the cows. These effects appeared additive, but 5% fat appeared to be excessive.

Milk yield and composition of lactating cows fed steam-flaked sorghum and graded concentrations of ruminally degradable protein.

To determine the effect of various amounts of ruminally undegradable protein in the diets of lactating cows fed steam-flaked sorghum, 24 Holstein cows (90 +/- 50 d in milk) were assigned to three treatments: 0.8% urea, 6% soybean meal, or 5% fish meal. Respective percentages of ruminally undegradable protein in the diets (as a percentage of crude protein) were 30, 35, and 39%. All diets contained 37% alfalfa hay; 3 to 5% cottonseed hulls; 10 to 13% whole cottonseed; 39% steam-flaked sorghum (360 g/L); 5% of a molasses, mineral, and vitamin supplement; and the different protein supplements. Intake of dry matter was higher for cows fed urea than for cows fed soybean meal or fish meal diets. In cows that yielded more than 40 kg/d of milk (4 cows per treatment), the soybean meal and fish meal diets resulted in higher yields of milk and 3.5% fat-corrected milk and a greater efficiency of conversion of feed to milk than did the urea diet. Cows that yielded less than 40 kg/d of milk (4 cows per treatment) at the beginning of treatment tended to yield more milk when fed urea than when fed the protein supplements. Nutrient digestibilities were not greatly affected by source of N, suggesting a beneficial effect of urea supplementation on nutrient digestibilities because replacement of protein supplements with cottonseed products caused the neutral detergent fiber content of the urea diet to be about 7% higher than that of the other diets. These data show that response to ruminally undegradable protein in diets of lactating cows fed steam-flaked sorghum was related to milk yield.

Influence of fat source and sorghum grain treatment on performance and digestibilities of high yielding dairy cows.

Forty-eight lactating Holstein cows averaging 81 d in milk were allotted to eight blocks based on milk yield during the 14-d pretreatment period and randomly assigned to six treatment groups in a 2 x 3 factorial arrangement of treatments for 64 d. Factors were type of sorghum grain processing [dry-rolled vs. steam-flaked; fed at 34% of dry matter (DM) in a total mixed ration (TMR) based on alfalfa] and type of supplemental fat (2.5% of DM as cottonseed oil, tallow, or prilled fatty acids). Compared with dry-rolled sorghum, steam-flaked sorghum did not affect milk yield, fat percentage, or fat yield but did increase milk protein percentage, body weight gains, and estimated net energy for lactation (22%). Fat source did not affect lactational response, but, compared with tallow, prilled fatty acids tended to decrease DM intake. Steam-flaked sorghum, compared with dry-rolled sorghum, increased digestibilities of DM, organic matter, crude protein, and starch, regardless of fat source. The TMR containing prilled fat had lower digestibilities of DM and organic matter than did TMR containing cottonseed oil or tallow; and TMR containing prilled fat had lower digestibilities of crude protein and total fatty acids than did TMR containing tallow. This study showed that steam-flaking of sorghum grain increased milk protein content, body weight gains, and estimated net energy for lactation, regardless of dietary fat source.

Comparison of barley and sorghum grain processed at different densities for lactating dairy cows.

To vary ruminally degradable starch, sorghum grain was dry-rolled or steam-flaked to different densities and compared with dry-rolled barley in total mixed diets fed to 40 lactating cows (111 d of lactation) assigned to five dietary treatments. Diets contained (percentage of dry matter) 35% alfalfa hay, 4.1% cottonseed hulls, 10% whole cottonseed, 2% fish meal, 4% soybean meal, and 4.9% of a molasses, mineral, and vitamin supplement. Treatments were 40% sorghum grain either dry-rolled or steam-processed at flake densities of 437, 360, and 283 g/L. A fifth diet containing 42% dry-rolled barley was fed. Cows were blocked according to pretreatment (14 d) milk yield and received experimental diets for 56 d. Increasing ruminal starch degradability by including steam-flaked sorghum grain or barley in the diet did not increase milk yield or milk protein percentage and yield, as was shown in eight previous studies. Steam-flaked sorghum or dry-rolled barley in the diet decreased dry matter intake, resulting in a 10 to 19% higher efficiency of conversion of feed dry matter to milk than that for dry-rolled sorghum. Milk urea N was decreased, and milk casein yield tended to be increased, by steam-flaking sorghum at the moderate density compared with dry-rolling or fine flaking. Dietary protein was more efficiently converted to milk protein and casein from flaked sorghum and dry-rolled barley than from dry-rolled sorghum. In this study, increasing ruminal starch degradability resulted in higher feed efficiency and lower feed intakes, bu optimal flake densities for steam-processed sorghum gain to maximize milk and milk protein yield were not clarified.

Influence of steam-flaked sorghum grain and supplemental fat on performance of dairy cows in early lactation.

Effects of sorghum grain processing and fat supplementation on milk yield and composition and nutrient digestibilities were studied. Thirty-six Holstein cows were assigned at 5 d of lactation for 91 d to treatment: 1) dry-rolled sorghum, no added fat; 2) dry-rolled sorghum plus Ca salts of long-chain fatty acids; 3) steam-flaked sorghum, no added fat; and 4) steam-flaked sorghum plus Ca salts of long-chain fatty acids. Fat increased ether extract in diets from 5.7 to 7.5% of DM. Milk yield was 4.0 kg/d (12%) higher for cows fed steam-flaked than dry-rolled sorghum. Fat tended to decrease milk yield because of lower DMI, and feed efficiencies increased 15%. Milk protein decreased .14% on dry-rolled sorghum with added fat, but only .01% on the steam-flaked diet. Milk protein yield was higher for cows fed flaked than rolled sorghum. Flaking decreased milk fat percentage, but not yield. Added fat decreased shorter chain fatty acids in milk (C4 to C14), but oleic acid increased. Flaking increased C10, C12, and C14, but decreased oleic acid. Digestibilities for DM, CP, starch, and ether extract were higher for flaked and fat-supplemented diets. Flanking of sorghum prevented the milk protein decrease reported for early lactation cows fed added fat.

Effect of source and amount of supplemental fat on lactation and digestion in cows.

Thirty Holstein cows averaging 50 DIM were assigned to five dietary treatments for 75 d to determine the effects of source and amount of supplemental fat on milk yield, milk composition, and nutrient digestibilities. Diets were 1) control, 2) diet 1 plus 12% whole cotton-seed, 3) diet 2 plus 2.2% safflower oil, 4) diet 2 plus 2.2% prilled tallow fatty acids, and 5) diet 2 plus 4.4% prilled tallow fatty acids. Milk yield was increased an average of 2.1 kg/d by addition of 2.2% prilled tallow fatty acids or safflower oil to the diet (7% fatty acids) containing 12% whole cottonseed. However, when fatty acids were increased to 9.1% with additional prilled tallow fatty acids, milk yield, DMI, and fatty acid digestibility decreased. Whole cottonseed alone and in combination with all fat additions decreased milk protein concentrations. Safflower oil increased C18:0, C18:1, and C18:2 fatty acids in milk. Digestibilities of OM, NDF, and ADF were not affected by diet. Supplementation of a saturated or unsaturated fat source to increase fatty acid content to 7.0% of dietary DM increased milk yield, but a further increase in fat to 9.1% with the saturated source appeared excessive for cows yielding 30 to 35 kg/d of milk.

Effect of three supplemental fat sources on lactation and digestion in dairy cows.

Tallow, Ca salts of palm fatty acids, or prilled fatty acids were added at 2.5% to a control diet (3.7% fatty acids) containing 7.2% ammoniated whole cottonseed. Diets were fed to midlactation cows (6 cows per treatment) for 72 d to determine effects of fat supplementation and fat source on lactation performance and nutrient digestibilities. Dry matter intake did not differ among treatments. Milk yield was 31.6 kg/d for the control and increased an average of 2.1 kg/d with fat supplementation. Source of supplemental fat did not significantly affect lactation performance. Added fat decreased milk protein content but did not affect protein yield. Milk fat, lactose, and SNF contents did not differ among treatments. Overall fat supplementation did not affect digestibilities of DM, ADF, or NDF but decreased digestibility of fatty acids. Contribution of de novo fatty acids to milk fat was decreased with fat supplementation. Addition of 2.5% fat to a diet containing a medium amount of fat from whole cottonseed increased milk yield. Tallow, Ca salt of palm fatty acids, and prilled fatty acids did not differ in milk yield response.