Relationship between the amino acid release kinetics of feed proteins and nitrogen balance in finishing pigs.

In current nutrition requirements of swine, although the protein diets are formulated based on the ileal digestibility of protein and amino acid (AA), there is a difference in nitrogen utilisation among various protein diets, which might be related to the AA release kinetics. To evaluate the relationship between AA release kinetics of feed proteins and nitrogen balance in finishing pigs, pigs were fed diets based on casein (CAS) or corn gluten meal (CGM) at normal or low-protein concentrations, and the AA release patterns were assessed. A 2 × 2 full factorial experimental design was used. 24 pigs (Duroc × Landrace × Yorkshire) with an initial weight of 67.0 ± 1.8 kg were randomly assigned to consume a normal-protein casein-based diet (N.CAS, 10% CP), normal-protein corn gluten meal-based diet (N.CGM, 10% CP), low-protein casein-based diet (L.CAS, 8.5% CP), or low-protein corn gluten meal-based diet (L.CGM, 8.5% CP) for 14 days (n = 6 per group; pigs housed and fed separately). The low-protein diets were associated with a more rapid release of AAs in the early stages of gastric digestion than the normal-protein diets. The N.CAS and L.CAS diets were associated with a peak AA release at approximately 4 h during trypsin digestion, whereas N.CGM and L.CGM were at approximately 16 h. The N.CAS diet was associated with the least dispersed release curves and lowest synchronisation indexes, implying that it was associated with the best AA release synchronism, whereas the L.CGM diet was on the contrary. The nitrogen intake (NI), faecal nitrogen, urine nitrogen (UN), total nitrogen, net protein utilisation and apparent biological value (ABV) of protein of pigs fed the L.CAS or L.CGM diets were lower than those fed the N.CAS or N.CGM diets (P < 0.05). Notably, there was a difference in NI (P < 0.05) and trends with respect to UN and ABV (0.05 < P < 0.1), but no differences in retained nitrogen or apparent nitrogen digestibility between pigs fed the N.CAS or L.CAS diets and those fed the N.CGM or L.CGM diets. Pigs fed the N.CAS or N.CGM diets had higher serum concentrations of UN than pigs fed the L.CAS or L.CGM diets (P < 0.05), but there were no differences in serum total protein, albumin, triglyceride, glucose, alanine transaminase, or aspartate aminotransferase between the groups. In addition, there was an interaction between protein level and protein source on serum globulin (P < 0.05). Therefore, the diet with a better AA release synchronism can improve protein utilisation efficiency in finishing pigs and to reduce environmental pollution.

[Advances in epidemiology and serum markers for the noninvasive diagnosis of nonalcoholic fatty liver disease].

The global prevalence rate of nonalcoholic fatty liver disease (NAFLD) has increased year by year, and it has become the number one cause for chronic liver disease in China. In addition, the trend of NAFLD has become more pronounced and evident in female gender and younger age group. The long-term persistence of fatty liver disease may cause serious consequences. There are no accepted diagnostic criteria for diagnosing noninvasive diagnosis of NAFLD. Alpha-ketoglutarate is a newly discovered serological marker of high diagnostic value and considered the most valuable potential biomarker along with cytokeratine-18 (CK-18).

[General plan of Shanghai Diet and Health Survey].

Shanghai Diet and Health Survey (SDHS) was designed to prospectively access local residents' food consumption, energy and nutrient intake, related chemical contaminant exposure, and the seasonal change trend to explore the relationship of diet with health. Data from SDHS can be used as fundamental information and scientific evidences for the development of local nutrition and food safety policies.

Effect of Preoperative Low Maximal Flow Rate on Postoperative Voiding Trials after the Midurethral Sling Procedure in Women with Stress Urinary Incontinence.

OBJECTIVES: To evaluate the effects of preoperative low maximal flow rate (Qmax) on voiding trials after the midurethral sling (MUS) procedure in women with stress urinary incontinence (SUI). METHODS: One hundred and sixty-eight women who underwent MUS procedure were enrolled. Preoperative free uroflowmetry was performed and patients were divided by Qmax. Low Qmax was defined as a Qmax under 15 mL/sec with voided volume at least 150 mL. Surgical results, failure of voiding trial, and postoperative uroflowmetry parameters were compared between the groups. Failure of voiding trial was defined by a PVR more than 100 mL on postoperative uroflowmetry. RESULTS: At the discharge day, there were 42 cases showing failure of voiding trial and 33 cases requiring CIC, but only one patient showed failure of voiding trial at 12 months postoperatively. Overall, 48 patients had preoperative low Qmax. Low Qmax group showed lower Qmax in all of postoperative uroflowmetry, but there were no significant differences in the rate of postoperative voiding trial failure or CIC. The low Qmax group was then divided into two groups according to the preoperative detrusor pressure at Qmax over and under 20 cmH2 O in pressure flow study. Comparing the two groups, no significant differences were observed in the cure rate, voiding trial failure or CIC. CONCLUSIONS: Our results suggest that women with preoperative low Qmax experienced no definite unfavorable voiding problem from the MUS procedure compared to those with normal voiding function. MUS procedure may be regarded as a safe and successful procedure in SUI women with low Qmax.

Resveratrol regulates mitochondrial reactive oxygen species homeostasis through Sirt3 signaling pathway in human vascular endothelial cells.

Mitochondrial reactive oxygen species (mtROS) homeostasis plays an essential role in preventing oxidative injury in endothelial cells, an initial step in atherogenesis. Resveratrol (RSV) possesses a variety of cardioprotective activities, however, little is known regarding the effects of RSV on mtROS homeostasis in endothelial cells. Sirt3 is a mitochondrial deacetylase, which plays a key role in mitochondrial bioenergetics and is closely associated with oxidative stress. The goal of the study is to investigate whether RSV could attenuate oxidative injury in endothelial cells via mtROS homeostasis regulation through Sirt3 signaling pathway. We found that pretreatment with RSV suppressed tert-butyl hydroperoxide (t-BHP)-induced oxidative damage in human umbilical vein endothelial cells (HUVECs) by increasing cell viability, inhibiting cell apoptosis, repressing collapse of mitochondrial membrane potential and decreasing mtROS generation. Moreover, the enzymatic activities of isocitrate dehydrogenase 2 (IDH2), glutathione peroxidase (GSH-Px) and manganese superoxide dismutase (SOD2) as well as deacetylation of SOD2 were increased by RSV pretreatment, suggesting RSV notably enhanced mtROS scavenging in t-BHP-induced endothelial cells. Meanwhile, RSV remarkably reduced mtROS generation by promoting Sirt3 enrichment within the mitochondria and subsequent upregulation of forkhead box O3A (FoxO3A)-mediated mitochondria-encoded gene expression of ATP6, CO1, Cytb, ND2 and ND5, thereby leading to increased complex I activity and ATP synthesis. Furthermore, RSV activated the expressions of phosphorylated adenosine monophosphate-activated protein kinase (p-AMPK), peroxisome proliferator-activated receptor gamma coactivator-1α (PGC-1α) and Sirt3, as well as estrogen-related receptor-α (ERRα)-dependent Sirt3 mRNA transcription, which were abolished in the presence of AMPK inhibitor and AMPK, PGC-1α or Sirt3 siRNA transfection, indicating the effects of RSV on mtROS homeostasis regulation were dependent on AMPK-PGC-1α-ERRα-Sirt3 signaling pathway. Our findings indicated a novel mechanism that RSV-attenuated oxidative injury in endothelial cells through the regulation of mtROS homeostasis, which, in part, was mediated through the activation of the Sirt3 signaling pathway.

Equol promotes rat osteoblast proliferation and differentiation through activating estrogen receptor.

Phytoestrogens have been suggested as alternative treatment for postmenopausal osteoporosis. Equol, a metabolite of daidzein, has been shown to inhibit bone loss in ovariectomized mice and rats. However, whether or not equol influences the formation of bone has not yet been investigated. Therefore, we investigated the effect of equol on the proliferation and differentiation of rat primary osteoblasts and explored the involved mechanisms. Different equol concentrations significantly promoted the proliferation of osteoblasts after 48- and 72-h incubations. The alkaline phosphatase (ALP) activity also increased significantly in all of the equol and 17ß-estradiol (E2) groups, except for the lowest (0.01 µM) equol group. Equol also significantly elevated the osteocalcin levels. The effects of equol on osteoblast proliferation, ALP activity, and osteocalcin levels were blocked by the estrogen receptor (ER) antagonist ICI182780. After a 24-h incubation, the expression of protein kinase C alpha (PKCα) in osteoblasts was significantly increased by equol. In conclusion, our study demonstrated that equol could promote the proliferation and differentiation of rat osteoblasts through activating the ER-PKCα-related signaling pathway, suggesting that equol could promote bone formation. These results suggest that equol could be a potential alternative agent for the management of postmenopausal osteoporosis.

NVP-BEZ235, a dual PI3K/mTOR inhibitor synergistically potentiates the antitumor effects of cisplatin in bladder cancer cells.

The PI3K/Akt/mTOR pathway is a prototypic survival pathway and constitutively activated in many malignant conditions. Moreover, activation of the PI3K/Akt/mTOR pathway confers resistance to various cancer therapies and is often associated with a poor prognosis. In this study, we explored the antitumor effect of NVP-BEZ235, a dual PI3K/mTOR inhibitor in cisplatin-resistant human bladder cancer cells and its synergistic interaction with cisplatin. A human bladder cancer cell line with cisplatin resistance was exposed to escalating doses of NVP-BEZ235 alone or in combination with cisplatin and antitumor effects was determined by the CCK-8 assay. Based on a dose-response study, synergistic interaction between NVP-BEZ235 and cisplatin was evaluated by combination index (CI), three-dimensional model and clonogenic assay. The combination of NVP-BEZ235 and cisplatin caused significant synergistic antitumor effect in cisplatin-resistant bladder cancer cells over a wide dose range and reduced the IC50 of NVP-BEZ235 and cisplatin by 5.6- and 3.6-fold, respectively. Three-dimensional synergy analysis resulted in a synergy volume of 388.25 µM/ml2% indicating a strong synergistic effect of combination therapy. The combination therapy caused cell cycle arrest and caspase-dependent apoptosis. Although NVP-BEZ235 suppressed PI3K/mTOR signaling without any paradoxical induction of Akt activity, it caused MEK/ERK pathway activation. The present study demonstrated that the PI3K/mTOR dual inhibitor NVP-BEZ235 can synergistically potentiate the antitumor effects of cisplatin in cisplatin-resistant bladder cancer cells though the suppression of cell cycle progression and the survival pathway as well as induction of caspase-dependent apoptosis.

Inhibition of the IKK/NF-κB pathway by AAV gene transfer improves muscle regeneration in older mdx mice.

The IκB kinase (IKKα, ß and the regulatory subunit IKKγ) complex regulates nuclear factor of κB (NF-κB) transcriptional activity, which is upregulated in many chronic inflammatory diseases. NF-κB signaling promotes inflammation and limits muscle regeneration in Duchenne muscular dystrophy (DMD), resulting in fibrotic and fatty tissue replacement of muscle that exacerbates the wasting process in dystrophic muscles. Here, we examined whether dominant-negative forms of IKKα (IKKα-dn) and IKKß (IKKß-dn) delivered by adeno-associated viral (AAV) vectors to the gastrocnemius (GAS) and tibialis anterior (TA) muscles of 1, 2 and 11-month-old mdx mice, a murine DMD model, block NF-κB activation and increase muscle regeneration. At 1 month post-treatment, the levels of nuclear NF-κB in locally treated muscle were decreased by gene transfer with either AAV-CMV-IKKα-dn or AAV-CMV-IKKß-dn, but not by IKK wild-type controls (IKKα and ß) or phosphate-buffered saline (PBS). Although treatment with AAV-IKKα-dn or AAV-IKKß-dn vectors had no significant effect on muscle regeneration in young mdx mice treated at 1 and 2 months of age and collected 1 month later, treatment of old (11 months) mdx with AAV-CMV-IKKα-dn or AAV-CMV-IKKß-dn significantly increased levels of muscle regeneration. In addition, there was a significant decrease in myofiber necrosis in the AAV-IKKα-dn- and AAV-IKKß-dn-treated mdx muscle in both young and old mice. These results demonstrate that inhibition of IKKα or IKKß in dystrophic muscle reduces the adverse effects of NF-κB signaling, resulting in a therapeutic effect. Moreover, these results clearly demonstrate the therapeutic benefits of inhibiting NF-κB activation by AAV gene transfer in dystrophic muscle to promote regeneration, particularly in older mdx mice, and block necrosis.

Age and sex dependent genetic effects of neuropeptide Y promoter polymorphism on susceptibility to ischemic stroke in Koreans.

BACKGROUND: In our previous study, the neuropeptide Y (NPY) C-399T promoter polymorphism (rs16147C>T) was identified as a risk factor for ischemic stroke in Koreans. In this study, we investigated whether age and sex modify the genetic effect of C-399T on susceptibility to ischemic stroke. METHODS: A total of 1,350 subjects (802 ischemic stroke patients, 548 healthy controls) were genotyped for C-399T using a primer extension method. The results were statistically analyzed for the genetic association of C-399T with ischemic stroke and clinical parameters. RESULTS: The TT genotype for C-399T was observed at a significantly lower frequency in stroke patients relative to control (CC+CT vs. TT, odds ratio [OR]=0.578, 95% confidence interval [95% CI]=0.360-0.927, P<0.05). This trend was also observed in female (OR=0.495, 95% CI=0.240-1.022) and older subjects (y>60, OR=0.556, 95% CI=0.304-1.018) with borderline statistical significance (P=0.0571 and P=0.0574, respectively). However, C-399T allele frequency was not different between controls and stroke patients in any groups. The C-399T polymorphism was found to be associated with body mass index and levels of some blood lipids. CONCLUSIONS: The C-399T NPY promoter polymorphism should be considered a genetic risk factor for ischemic stroke in the older adult and female Korean populations.

Association of the C-399T promoter polymorphism of neuropeptide Y with susceptibility to ischemic stroke.

OBJECTIVES: The common sequence variants of neuropeptide Y (NPY) were known to be associated with some kinds of diseases including stroke. This study investigated the association of NPY promoter polymorphism, C-399T, with ischemic stroke and its underlying mechanism using in vitro systems. DESIGN AND METHODS: Study subjects consisted of 444 ischemic stroke patients and 326 controls without stroke. C-399T genotyping was conducted by a primer extension-based method. Plasma NPY was quantified with an enzyme immunoassay, and transcription characteristics were investigated by a reporter gene assay and an enzyme mobility shift assay. RESULTS: A significantly lower frequency of TT genotype was observed in a stroke group (OR[95% CI], 0.399[0.187-0.854], p=0.0180). The C-399T polymorphism affected the transcription efficiency of NPY gene and its genotypes were related to the changes in plasma NPY levels. CONCLUSION: This study suggests that NPY promoter polymorphism, C-399T, should be considered a genetic risk factor for ischemic stroke.

Effects of ipt gene expression on the physiological and chemical characteristics of Artemisia annua L.

An isopentenyl transferase gene (ipt) from T-DNA was transferred into Artemisia annua L. via Agrobacterium tumefaciens. The ipt gene was placed in a binary vector under the control of the CaMV 35S promoter. Leaf explants were infected with A. tumefaciens LBA4404 containing pBIipt to induce the buds. Nineteen shoot lines were selected, which were resistant to kanamycin. Polymerase chain reactions and Southern blotting confirmed that at least five shoot lines contained the foreign gene. The results of RT-PCR and Northern blotting analyses suggested that the foreign ipt gene of the transgenic shoot was expressed. Cytokinins, chlorophyll and artemisinin contents were found increased at different degree. Content of cytokinins (iPA and iP) was elevated 2- to 3-fold, chlorophyll increased 20-60% and artemisinin increased 30-70% compared with the control plants, respectively. A direct correlation was found between the contents of cytokinins, chlorophyll and artemisinin. This may be the first report on the relationship between endogenous cytokinin content and the production of secondary metabolites in plants.

[Effect of cholesterol deficiency on the membrane fluidity of Jurkat T lymphocytes].

The mechanism of cholesterol deficiency on inhibiting the proliferation of T lymphocytes was investigated on cell membrane fluidity by using fluorescence polarization measurement, 3H-TdR incorporation test and flow cytometer analysis in Jurkat cells, in order to clarify the importance of cholesterol in maintaining the normal function of lymphocytes. The results showed that cell membrane fluidity was increased, cell proliferation was inhibited and blocked on G0/G1 phase in Jurkat cells after being cultured with lovastatin (an inhibitor of rate limiting enzyme for cholesterol synthesis) for 3 days. When the cells were treated with LDL. The changes could be partially buffered. These results suggest that the change of membrane fluidity may be caused by cholesterol deficiency on the proliferation of Jurkat cells.

[Influences of taurine and micronutrients on nitric oxide synthase expression and cGMP content in rat retina].

AIM: To investigate the influence of taurine and micronutrients on visual signal transmission. METHODS: Wistar rats were divided into three groups, that is control group, experiment group 1 and experiment group 2, and fed for 3 weeks with normal diet, 5 times and 10 times doses of requirements of taurine, vitamin A, vitamin B, zinc and selenium, then each treatment group were divided into light group and dark adaptation group. After feeding another 3 days in different environments with normal diet, all animals were killed and cGMP level and NOS expression were analysed in retina and retinogeniculate. RESULTS: The NOS expression and cGMP contents of photoreceptor cells, visual cortex and retinogeniculate were increased in dark adaptation group compared with light group. Nutritional intervention could enhance the NOS staining in dark environment, increased the cGMP contents whether light or dark condition. CONCLUSION: The distribution, expression and content of NO and cGMP are quite different in various light adaptation status. Taurine and micronutrient intervention may modurate the visual signal transmission or vision function mediated by the changes of NO or cGMP.

[Study of the role of cyclin-dependent kinases (CDKs) in retinoic acid (RA) inducing HL-60 cell differentiation].

OBJECTIVE: To investigate the role of CDKs in the RA inducing HL-60 cell differentiation. METHODS: The effects of all-trans retinoic acid (ATRA) and arotinoid ethylester(AE) at the dosage of 5 x 10(-6) mol/L on the proliferation and differentiation of HL-60 cells were examined by NBT reduction test and cytometry analysis. Meanwhile, histone H1 kinase assay was used to observe the changes of CDK2 activities in HL-60 cells treatment with ATRA and AE. The amounts of both cyclin E/CDK2 and cyclin D1/CDK4 complexes were observed with immunoprecipitates. RESULTS: Both ATRA and AE inhibited the growth of HL-60 cells, arrested cells in G1/G0 phase, and induced cells to differentiation. The activities of cyclin E/CDK2 and the amounts of cyclin D1/CDK4 complexes were decreased in ATRA- or AE-treated HL-60 cells. CONCLUSION: The effects of RA on the proliferation and differentiation of HL-60 cells may be associated with significant decreases in the activities of CDKs.

Obesity in youth and middle age and risk of colorectal cancer in men.

To investigate an association between colon cancer and obesity during early adulthood--a potentially important period in the etiology of this disease--the authors assembled, by computer linkage, a population-based historical cohort of 52,539 men born between 1913 and 1927 residing in Hawaii (USA), for whom weight and height had been recorded in 1942-43 and 1972. Linkage of this cohort to the Hawaii Tumor Registry resulted in the identification of 737 incident cases of colorectal cancer for 1972-86. An average of 3.8 cancer-free controls were matched to each case on month and year of birth and ethnicity of the parents. A case-control analysis in each anatomic subsite of the large bowel revealed that both early and middle-age body mass increased the risk of sigmoid cancer in men in a dose-dependent fashion. The odds ratios (OR) for sigmoid cancer for the highest compared with the lowest tertiles of Quetelet index were: 2.1 (95 percent confidence interval [CI] = 1.4-3.2) and 1.7 (CI = 1.1-2.5), at ages 15-29 and in prediagnostic years, respectively. These associations were additive and independent of socioeconomic status. Men who were above the median Quetelet index in 1942 and 1972 had an OR of 2.7 (CI = 1.8-4.0), compared with those who were below the median in both periods. This study provides further evidence for an association of obesity with colon cancer in men and suggests that this association is limited to the sigmoid colon and may be related to both early and late events of colon carcinogenesis.

Early-age body size, adult weight gain and endometrial cancer risk.

To further characterize the association of obesity and endometrial cancer, in particular with regard to the role of early-age obesity and adult weight gain, the authors assembled by computer linkage a population-based historical cohort of 30,266 women born between 1913 and 1932, for whom weight and height had been recorded in 1942-43 and 1972. Linkage of this cohort to the Hawaii Tumor Registry resulted in the identification of 214 (mainly post-menopausal) incident cases of endometrial cancer for 1972-1986. An average of 37 cancer-free controls were matched to each case on month and year of birth and ethnicity. A case-control analysis, conducted in each 5-year birth cohort, revealed no clear association of endometrial cancer with weight, height or body mass at ages 10 to 29 years. However, positive associations with adult body weight and gain in body mass since 1942 were observed for women diagnosed at age 60 or older. This association with obesity was strongest in women whose body mass was below the median in 1942 and equal to or above the median in 1972. No association with body size was detected in women diagnosed before age 60. Parity, age at first birth and socioeconomic indicators for 1942 and 1972 did not confound the analysis. These findings suggest that obesity affects the late stages of endometrial carcinogenesis, and the possibility that one or more determinants of weight gain may be independently associated with endometrial cancer risk.

Body size at different periods of life and breast cancer risk.

The inconclusive findings of past analytic epidemiologic studies on diet and breast cancer may have resulted from the inability of these studies to assess early dietary exposures. The role of macronutrient intake during early life can be indirectly studied, however, by examining past and present body size. The authors identified by computer linkage a population-based historical cohort of 38,084 women born between 1918 and 1943, on whom information about weight and height had been recorded in Hawaii in both 1942-1943 and 1972. Linkage of this cohort to the Hawaii Tumor Registry resulted in the identification of 607 incident cases of breast cancer for 1972-1983. An average of 4.4 cancer-free controls were matched to each case on year and month of birth and race of the parents. A matched case-control analysis, conducted in each five-year birth cohort, revealed a negative association of adolescent body mass to premenopausal breast cancer. This negative association was statistically significant in girls aged 10-14 years in 1942 (p for trend, 0.004), was present in all ethnic groups, and was strongest among overweight young women who remained overweight in adulthood. Early-age weight, height, and body surface area were not associated with either pre- or postmenopausal breast cancer. Adult weight and gain in body mass since 1942 were positively associated with postmenopausal breast cancer risk. Adjustment for age at first birth, parity and socioeconomic indicators for 1942 and 1972 did not modify the results. This study provides evidence for a protective role of adolescent obesity against premenopausal breast cancer, and for an enhancing role of a positive energy balance during adult life on postmenopausal breast cancer.

Digital dermal patterns in breast cancer.

Fingerprints of 570 breast cancer cases and the same number of matched controls were obtained from the population-based finger print file in Hawaii for studying the association between breast cancer and digital dermal patterns and ridge counts. The results showed that breast cancer patients had a significant excess of radial loops on the left hand. It was also found that the frequency of ulnar loops on the left hand was significantly elevated for premenopausal women with breast cancer, whereas an excess of radial loops on the left hand was observed for the postmenopausal women with breast cancer. No significant difference for the total and absolute ridge counts was found between breast cancer patients and controls.