12-d Treatment Research Articles

Discovery of 4-((3,4-dichlorophenyl)amino)-2-methylquinolin-6-ol derivatives as EGFR and HDAC dual inhibitors

In patients with non-small cell lung cancer (NSCLC), the standard therapy consists of selective tyrosine kinase inhibitors that target epidermal growth factor receptors (EGFR). Nonetheless, their clinical utility is primarily limited by the development of resistance to drugs. HDAC inhibitors have been shown in studies to reduce the level of EGFR that is expressed and downregulate the EGFR-induced phosphorylation of AKT and ERK. Therefore, dual inhibitors of EGFR and HDAC provide a potential approach as combination treatment synergistically inhibited the growth of NSCLC. Herein, we examined the EGFR inhibition effect of twenty compounds which designed and synthesized by us previously. Among them, compounds 12c and 12d exhibited powerful antiproliferative activity against the NCI–H1975 cell line with IC50 values of 0.48 ± 0.07 and 0.35 ± 0.02 μM, correspondingly. In cell-free kinase assays, both 12c and 12d demonstrated target-specific EGFR inhibition against wild type (EGFRwt). Furthermore, the expression of EGFR and phosphorylation of the EGF-induced pathways were significantly suppressed under the treatment of 12c and 12d. Besides, both histones H3 and H4 exhibited increased levels of acetylation following 12c and 12d treatment. The animal experiments shown that 12d could prevent the growth of tumor, inhibited the expression of EGFR and the phosphorylation levels of p70 S6K, AKT and p38 MAPK in vivo, and did not cause organ damage to the mice during the experiment.Overall, the results illustrated that compound 12c and 12d could serve as effective EGFR and HDAC dual inhibitors in NSCLC cells. Our work offers an alternative strategy for NSCLC therapy.

Effect of photoperiod during incubation on embryonic temperature, hatch traits, and performance of 2 commercial broiler strains

Provision of light during incubation has shown the potential to enhance hatching traits and affect posthatch productivity, physiology, and behavior. In this study, 2 repeated trials were conducted to investigate the effect of photoperiod and strain on the embryo temperature, hatching traits and posthatch growth performance of 2 commercial strains of broilers (Ross 308 and Cobb 500). In each trial, hatching eggs were randomly distributed into 6 incubators with 3 photoperiod treatments: blue LED light for 12 h d-1 (12L:12D) or 18 h d-1 (18L:6D) during entire incubation were compared with no illumination condition (DARK). Data were analyzed as a 3×2 factorial arrangement with the trial as the blocking factor. Embryos incubated under 12L:12D and 18L:6D had lower air cell temperature (P < 0.05) than the DARK embryos from d 13 of incubation onward except on the day of candling. The response of air cell temperature to periodic illumination differed between 2 strains. Cobb embryos had lower air cell temperature in 12L:12D than those incubated with 18L:6D from d 16 of incubation onward, whereas lower air cell temperature was found in Ross embryos when illuminated with 18L:6D photoperiod compared to those under 12L:12D. The 12L:12D treatment was associated with improved (P < 0.05) navel closure condition of hatchlings. There were no differences in hatchability, embryo mortality, body weight, or length at hatch among photoperiod groups or its combination with strain. No differences in production parameters were found between DARK and illuminated groups. However, 12L:12D had heavier (P < 0.05) body weight on d 14 of age and higher (P < 0.05) body weight gain than 18L:6D from d 7 to 14 of age. The results of this study indicate that providing blue LED light up to 18 h d-1 has no detrimental effect on production of broilers, however, 12L:12D light regime improved chick quality at hatch compared to DARK and resulted in heavier birds by d 14 compared to 18L:6D.

Effects of different photoperiod conditions on survival, growth, and gonadal development of Takifugu rubripes adults

Photoperiod has a significant influence on fish. In this study, 2.5-year-old Takifugu rubripes (tiger puffer) adults were selected to evaluate the photoperiod impacts (8 h light (L):16 h dark (D), 12L:12D, 16L:8D) from September to January at 15–17 °C on their growth, gonadal development, and survival for a 100-day. The mortality rate of males and females tiger puffer differed among the different photoperiods, and the 8L:16D photoperiod was not suitable for survival of males. At the end of the experiment, the body length, weight gain rate, specific growth rate and condition factor of both males and females tiger puffer were similar among the three photoperiods (P > 0.05). Higher percentage of females had developed to later developing phase and males had developed to spawning capable phase in the 12L:12D treatment, respectively. In particular, 100% of males had testis at the spawning capable phase. The gonadosomatic index values of males and females in the 12L:12D group were significantly higher than those in the 16L:8D group (P < 0.05), but no significant differences between the 8L:16D and 12L:12D groups (P > 0.05). The serum testosterone (T) content of males and the serum estradiol-17β (E2) content of females were highest in the 12L:12D group. The expression of aralkylamine N-acetyltransferase 1a (aanat1a) in the diencephalon/midbrain (D/M) was elevated under 12L:12D treatment in both sex (P < 0.05). The expression level of iodothyronine deiodinase type 2 (dio2) in the saccus vasculosus in males was elevated under the 12L:12D photoperiod (P < 0.05). The levels of gonadotropin-releasing hormone 1 (gnrh1) and gnrh3 mRNA in the D/M of females at later developing stages in the 12L:12D treatment were significantly higher than those in of other treatment groups, and the level of gnrh3 in males under this photoperiod was significantly elevated (P < 0.05). The levels of gonadotropin-inhibiting hormone (gnih) mRNA under 12L:12D were significantly elevated in both males and females (P < 0.05). Under this photoperiod, the levels of follicle stimulating hormone b-subunit (fshb) and luteinizing hormone b-subunit (lhb) in the pituitary gland of females at later developing stages were significantly higher than those of the other two groups (P < 0.05). The expression levels of lhb in males in the 12L:12D and 8L:16D photoperiods were significantly higher than that at 16L:8D (P < 0.05). These results indicated that culture under the 12L:12D photoperiod from September prior to the spawning season could result in accelerated gonadal development of both male and female tiger puffer.

Preliminary evaluation of the green microalga Dunaliella salina as a potential feedstock for biodiesel: effect of molasses on growth and lipid profile

This research aims to evaluate the effect of molasses as a source of organic carbon on the growth and lipid profile of Dunaliella salina cultivated under different light regimes to determine its strategy and potential use to produce biodiesel. Two sources of nutrients (F/2 medium, F/2; molasses, M) and two light conditions (12 h light: 12 h darkness, 12L:12D) conformed four treatments (F/2 12L:12D, F/2 D, M 12L:12D, and MD) that were investigated with three replicates each. The concentration of microalgae produced by the groups under light conditions was more than double that of those grown in darkness. The highest biomass produced (46.91 ± 6.47 g L -1 ) was obtained in the F/2 medium with 12L:12D treatment, while the lowest (18.99 ± 0.78 g L -1 ) was in the M 12L:12D group. Regardless of the culture conditions, the lipids were mainly composed of the C16:0 and C18 fatty acids. The use of molasses as a culture medium produced the highest amount of C18:1, regardless of the light regime. Molasses promoted the highest production of fatty acids methyl esters. Depending on the culture medium, D. salina cells showed different percentages of saturated (SFA), monounsaturated (MFA), and polyunsaturated (PUFA) fatty acids. The highest amount of saturated fatty acids was obtained in F/2 12L:12D. The highest amount of polyunsaturated fatty acids was presented in the mixotrophic culture (molasses and light). The cultures grown with molasses were rich in monounsaturated fatty acids. An inexpensive industrial by-product, Cane molasses could be a low-cost feedstock for biodiesel in cultivating D. salina .

Se transformation and removal by a cattail litter treatment system inoculated with sulfur-based denitrification sludge: Role of the microbial community composition under various temperature and aeration conditions

With a narrow margin between deficiency and toxicity, rising levels of selenium (Se) are threatening aquatic ecosystems. To investigate the role of microorganisms in Se bioremediation, a cattail litter system inoculated with the sulfur-based denitrification sludge was conducted. The results show the litter, as a carrier and nutrient source for bacteria, efficiently removed Se by ~ 97.0% during a 12-d treatment with water circulating. As the major removal pathways, immobilization rates of selenite were ~ 2.9-fold higher than selenate, and the volatilization, contributing to ~ 87.7% of the total Se removal, was significantly correlated with temperature (positively) and oxidation-reduction potential (ORP; negatively). Using X-ray absorption spectroscopy to speciate litter-borne Se, more Se0 formed without aeration due to abundant Se-reducing bacteria, among which Azospira and Azospirillum were highly related to the removal of both Se oxyanions, while Desulfovibrio, Azoarcus, Sulfurospirillum, Thauera, Geobacter, Clostridium, and Pediococcus were the major contributors to selenate removal. Overall, our study suggests microbial Se metabolism in the litter system was significantly affected by temperature and ORP, which could be manipulated to enhance Se removal efficiency and the transformation of selenate/selenite into low toxic Se0 and volatile Se, reducing risks posed by the residual Se in the system.

Effects of Photoperiod Manipulation on Gonadal Activity of the Damselfish, Chromis notata.

ABSTRACTThis study investigated the effect of different photoperiods (14L: 10D, 12L:12D and 10L:14D) on the gonadal development and GtH mRNA expression in the pituitary of damselfish. The results showed that gonadosomatic index (GSI) was significantly lower in shot photoperiod (10L:14D), in comparison with other photoperiodic group during the spawning season. After 60 days treatment, histological analysis of gonad tissue showed that the gonad of 10L:14D and 12L: 12D treatment groups were resting phase with spermatogonia and perinucleolus stage oocytes but the gonad of 14L:10D treatment group was still ripe phase with spermatozoa and mature stage oocyte. The FSHβ and LHβ mRNA expression in pituitary drastically decreased shot photoperiod treatment from July (spawning period). These results suggest that photoperiod is considered to be the most effective environmental factor in controlling the reproductive cycle of damselfish.

Acupuncture combined with western medicine for CP/CPPS:a randomized controlled trial

To compare the clinical efficacy differences among acupuncture combined with western medicine, acupuncture alone and western medicine alone for chronic prostatitis/chronic pelvic pain syndrome (CP/CPPS). Ninety patients were randomly assigned into a needle-medicine group, an acupuncture group and a western medicine group, 30 patients in each group. The patients in the needle-medicine group were treated with acupuncture combined with western medicine; the scalp points included Shenting (GV 24), Xinhui (GV 22), Qianding (GV 21), Baihui (GV 20), Chengguang (BL 6), Tongtian (BL 7), etc. The body points were Zhongji (CV 3), Guanyuan (CV 4), Pangguangshu (BL 28), Ciliao (BL 32), etc. The acupuncture was given 30 min per treatment, once a day. Besides, oral administration of 0.2g levofloxacin (twice per day) and 0.2 mg tamsulosin (once a day) was applied. The patients in the acupuncture group and western medicine group were treated by acupuncture and western medicine respectively. 12-d treatment was taken as one session, and totally 2 sessions were given. The clinical efficacy of the three groups after treatment was compared as well as the National Institutes of Health-Chronic Prostatitis Symptom Index (NIH-CPSI) total score and pain score, self-rating anxiety scale (SAS) and self-rating depression scale (SDS) before and after treatment. During the trial two patients dropped out, as a result, 30 patients in the needle-medicine group, 29 patients in the acupuncture group and 29 patients in the western medicine group were included in the analysis. After treatment, 21 patients were cured, 6 patients were markedly effective, 2 patients were effective and 1 patient failed in the needle-medicine group;12 patients were cured, 10 patients were markedly effective, 5 patients were effective and 2 patients failed in the acupuncture group; 11 patients were cured, 12 patients were markedly effective, 4 patients were effective and 2 patients failed in the medicine group; the efficacy in the needle-medicine group was superior to those in the acupuncture group and medicine group (both P<0.05). Each score was improved after treatment in each group (all P<0.01); the total score of NIH-CPSI as well as SAS and SDS scores in the needle-medicine group were superior to those in the acupuncture group and medicine group (P<0.05, P<0.01); the pain scores of NIH-CPSI in needle-medicine group and acupuncture group were superior to that in the medicine group (P<0.05, P<0.01), but the difference between the needle-medicine group and acupuncture group was not significant (P>0.05). The efficacy of acupuncture combined with western medicine for CP/CPPS is superior to that of acupuncture alone and western medicine alone, which could improve the symptom of prostatitis as well as status of anxiety and depression.

The effect of dietary hydroxyproline and dietary oxalate on urinary oxalate excretion in cats1

In humans and rodents, dietary hydroxyproline (hyp) and oxalate intake affect urinary oxalate (Uox) excretion. Whether Uox excretion occurs in cats was tested by feeding diets containing low oxalate (13 mg/100 g DM) with high (Hhyp-Lox), moderate (Mhyp-Lox), and low hyp (Lhyp-Lox) concentrations (3.8, 2.0, and 0.2 g/100 g DM, respectively) and low hyp with high oxalate (93 mg/100 g DM; Lhyp-Hox) to 8 adult female cats in a 48-d study using a Latin square design. Cats were randomly allocated to one of the four 12-d treatment periods and fed according to individual energy needs. Feces and urine were collected quantitatively using modified litter boxes during the final 5 d of each period. Feces were analyzed for oxalate and Ca, and urine was analyzed for specific density, pH, oxalate, Ca, P, Mg, Na, K, ammonia, citrate, urate, sulfate, and creatinine. Increasing hyp intake (0.2, 2.0, and 3.8 g/100 g DM) resulted in increased Uox excretion (Lhyp-Lox vs. Mhyp-Lox vs. Hhyp-Lox; P < 0.05), and the linear dose-response equation was Uox (mg/d) = 5.62 + 2.10 × g hyp intake/d (r(2) = 0.56; P < 0.001). Increasing oxalate intake from 13 to 93 mg/100 g DM did not affect Uox excretion but resulted in an increase in fecal oxalate output (P < 0.001) and positive oxalate balance (32.20 ± 2.06 mg/d). The results indicate that the intestinal absorption of the supplemental oxalate, and thereby its contribution to Uox, was low (5.90% ± 5.24%). Relevant increases in endogenous Uox excretion were achieved by increasing dietary hyp intake. The hyp-containing protein sources should be minimized in Ca oxalate urolith preventative diets until their effect on Uox excretion is tested. The oxalate content (up to 93 mg/100 g DM) in a diet with moderate Ca content does not contribute to Uox content.

The effects of light stimulation during incubation on indicators of stress susceptibility in broilers

Providing light during incubation has been shown to decrease bilateral physical asymmetry of broilers posthatch, which may indicate that early light stimulation reduces later stress susceptibility. This experiment evaluated the effects of lighting during embryogenesis on other measures of stress responsiveness in broilers. Cobb 500 eggs (n = 1,404) were incubated under 0L:24D, 1L:23D, 6L:18D, or 12L:12D; the light level was 550 lx. The broilers were then raised in floor pens under a 12L:12D lighting regimen, and various stress parameters were measured during wk 3 to 6 of age. There was an effect of incubation lighting regimen on posthatch stress responses. Following 1 h of crating, the change in corticosterone (CORT) concentration was less in the 12L:12D (−0.06 ng/mL, P < 0.05) treatment than all other treatments (pooled mean = 0.24 ng/mL); however, there were no treatment differences in CORT response to adrenocorticotropic hormone administration (pooled mean pre- vs. 1 h postadministration = 17.5 ng/mL, P > 0.05). Anti-keyhole limpet hemocyanin titers were higher in the 12L:12D birds (92,395 units/mL; P < 0.05) than all other treatments (pooled mean = 68,407 units/mL) on d 1 postcrating. Additionally, composite asymmetry scores were lower in the 12L:12D treatment (0.92 mm) than all other treatments (pooled mean = 1.14 mm, P < 0.05). These results demonstrate that providing 12 h of light per day during incubation can reduce the stress susceptibility of broilers posthatch.

Administration of Minor Polar Compound-Enriched Extra Virgin Olive Oil Decreases Platelet Aggregation and the Plasma Concentration of Reduced Homocysteine in Rats1,

We investigated the effect of extra virgin olive oil (EVOO) on platelet aggregation and plasma concentrations of homocysteine (Hcy) redox forms in rats in relation to the minor polar compound (MPC) concentration of EVOO. We used 3 olive oil samples with similar fatty acid but different MPC concentrations: refined olive oil (RF) with traces of MPC (control oil), native EVOO with low MPC concentration (LC), and EVOO with high MPC concentration (HC) enriching LC with its own MPC. Oil samples were administered to rats by gavage (1.25 mL/kg body weight) using 2 experimental designs: acute (24-h food deprivation and killed 1 h after EVOO administration) and subacute (12-d treatment, a daily dose of oil for 12 d, and killed after 24 h of food deprivation). Platelet aggregation was induced by ADP (ex vivo tests) and a reduction in platelet reactivity occurred in cells from rats given LC in the subacute study and in cells from rats administered HC in both studies as indicated by an increase in the agonist half maximal effective concentration. HC inhibited platelet aggregation induced by low ADP doses (reversible aggregation) in cells of rats in both the acute and subacute studies, whereas LC had this effect only in the subacute experiment. Moreover, in rats administered HC in both experiments, the plasma concentration of free reduced Hcy (rHcy) was lower and Hcy bound to protein by disulfide bonds (bHcy) was greater than in RF-treated rats. bHcy was also greater in rats given LC than in RF-treated rats in the subacute experiment. Plasma free-oxidized Hcy was greater in rats given LC and HC than in those administered RF only in the subacute experiment. In conclusion, these results show that MPC in EVOO inhibit platelet aggregation and reduce the plasma rHcy concentration, effects that may be associated with cardiovascular protection.

Activation of SRC Kinase and Phosphorylation of Signal Transducer and Activator of Transcription-5 Are Required for Decidual Transformation of Human Endometrial Stromal Cells

Progesterone induces decidual transformation of estrogen-primed human endometrial stromal cells (hESCs), critical for implantation and maintenance of pregnancy, through activation of many signaling pathways involving protein kinase A and signal transducer and activator of transcription (STAT)-5. We have previously shown that kinase activation of v-src sarcoma (Schmidt-Ruppin A-2) viral oncogene homolog (SRC) kinase is closely associated with decidualization and that SRC is indispensable for maximal decidualization in mice. To address whether SRC kinase activity is essential for decidualization in humans, hESCs were infected with adenoviruses carrying enhanced green fluorescent protein alone (Ad-EGFP), a kinase-inactive dominant-negative mutant (Ad-SRC/K295R), or an inactive autophosphorylation site mutant (Ad-SRC/Y416F). The cells were cultured in the presence of estradiol and progesterone (EP) to induce decidualization and subjected to RT-PCR, immunoblot, and ELISA analyses. Ad-EGFP-infected hESCs exhibited decidual transformation and up-regulation of decidualization markers including IGF binding protein 1 and prolactin in response to 12-d treatment with EP. In contrast, hESCs infected with Ad-SRC/K295R remained morphologically fibroblastoid without production of IGF binding protein 1 and prolactin even after EP treatment. Ad-SRC/Y416F displayed similar but less inhibitory effects on decidualization, compared with Ad-SRC/K295R. During decidualization, STAT5 was phosphorylated on tyrosine 694, a well-known SRC phosphorylation site. Phosphorylation was markedly attenuated by Ad-SRC/K295R but not Ad-EGFP. These results indicate that the SRC-STAT5 pathway is essential for decidualization of hESCs.

Effects of photoperiod on growth and spawning efficiency of Nile tilapia (Oreochromis niloticus L.) broodstock in a recycling system

This study was conducted to evaluate the effects of photoperiod on the growth, feed conversion ratio (FCR) and spawning performance of Nile tilapia (Oreochromis niloticus) broodstock. Duplicate groups of two males (66±3.04 g) and six females (50.5±1.58 g) were stocked in 0.4 m3 fibreglass tanks in a recirculating water system at a male:female ratio of 1:3. The fish were subjected to four photoperiod treatments: 24:0 light:dark (L:D), 18L:6D, 12L:12D and 6L:18D. Light intensity was kept constant at about 2500 lx throughout the study. The fish were fed a commercial tilapia diet (35% crude protein and 16.6 MJ GE kg−1) at a daily rate of 2–3% of tank biomass, twice a day, for 130 days. Males grew significantly faster than females under all photoperiod regimes. The best growth rate and FCR were attained at 18L:6D, followed by 24L:0D, 12L:12D and 6L:18D respectively. The number of eggs per female, number of eggs per spawn and number of spawnings per female were all significantly higher in the 12L:12D treatment than in all other photoperiod cycles. Interspawning intervals and days elapsed per spawn were also shorter in the 12L:12D treatment. The time to first spawning was slightly longer in the 24L:0D and 6L:18D than in 12L:12D and 18L:6D light phases. The 18L:6D and 6L:18D photoperiods produced the lowest spawning performance. It is concluded that a 12L:12D photoperiod regime should be adopted for maximum fecundity, seed production and spawning frequencies of Nile tilapia broodstock reared in intensive, recirculating systems. If maximum reproduction is desired, a near-natural day length photoperiod should be used.

Termination of gonadal refractoriness in Turkish hamsters, Mesocricetus brandti.

The Turkish hamster (Mesocricetus brandti) is a photoperiodic species. In this investigation, we characterized the photoperiodic requirements for termination of gonadal refractoriness, defined as the inability of the animal to respond to short-day treatment with gonadal regression. Paired testes weights were reduced to less than 20% of their original weight by 10 wk of 12L:12D treatment. This was followed by spontaneous testicular recrudescence (completed by Week 25 of 12L:12D treatment), the overt indication of refractoriness to short photoperiods. Next, the period of long-day exposure sufficient for termination of refractoriness was determined. Refractory males were exposed to 16L:8D for 5 to 20 wk. Ten weeks of 16L:8D treatment was enough for the animals to regain the sensitivity to a second challenge of 12L:12D treatment. Fifteen weeks of 20L:4D or 16L:8D terminated refractoriness in female Turkish hamsters; 20L:4D therefore was not interpreted as a short day by refractory hamsters. This was unexpected because in photosensitive animals this photoperiod acts like a short day, causing gonadal regression. These results suggest that Turkish hamsters are similar to Syrian hamsters in that both species require two or more months of long days in summer to recover sensitivity to the short days of the following fall.