MicroRNAs as biomarkers for major depression: a role for let-7b and let-7c.

There is a growing emphasis in the field of psychiatry on the need to identify candidate biomarkers to aid in diagnosis and clinical management of depression, particularly with respect to predicting response to specific therapeutic strategies. MicroRNAs are small nucleotide sequences with the ability to regulate gene expression at the transcriptomic level and emerging evidence from a range of studies has highlighted their biomarker potential. Here we compared healthy controls (n=20) with patients diagnosed with major depression (n=40) and who were treatment-resistant to identify peripheral microRNA biomarkers, which could be used for diagnosis and to predict response to electroconvulsive therapy (ECT) and ketamine (KET) infusions, treatments that have previously shown to be effective in treatment-resistant depression (TRD). At baseline and after treatment, blood samples were taken and symptom severity scores rated using the Hamilton Depression Rating Scale (HDRS). Samples were analyzed for microRNA expression using microarray and validated using quantitative PCR. As expected, both treatments reduced HDRS scores. Compared with controls, the baseline expression of the microRNA let-7b was less by ~40% in TRD patients compared with controls. The baseline expression of let-7c was also lower by ~50% in TRD patients who received ECT. Bioinformatic analysis revealed that let-7b and let-7c regulates the expression of 27 genes in the PI3k-Akt-mTOR signaling pathway, which has previously been reported to be dysfunctional in depression. The expression of miR-16, miR-182, miR-451 and miR-223 were similar to that in controls. Baseline microRNA expression could not predict treatment response and microRNAs were unaffected by treatment. Taken together, we have identified let-7b and let-7c as candidate biomarkers of major depression.

Characterization of the feeding inhibition and neural activation produced by dorsomedial hypothalamic cholecystokinin administration.

Within the dorsomedial hypothalamus (DMH), cholecystokinin (CCK) has been proposed to modulate neuropeptide Y (NPY) signaling to affect food intake. However, the neural circuitry underlying the actions of this CCK-NPY signaling system in the controls of food intake has yet to be determined. We sought to characterize the feeding inhibition and brain neural activation produced by CCK administration into the DMH of rats. We determined the time course of feeding inhibitory effects of exogenous DMH CCK, assessed NPY gene expression in the DMH in response to DMH CCK administration, and characterized c-Fos activation in the entire brain induced by CCK injection into the DMH using c-Fos like immunohistochemistry. We found that parenchymal injection of CCK into the DMH decreased food intake during the entire 22 h observation period, with a primary effect in the first 4 h, and down-regulated NPY gene expression in the DMH. c-Fos immunohistochemistry revealed that DMH CCK increased the number of c-Fos positive cells in the paraventricular nucleus (PVN), arcuate nucleus, suprachiasmatic nucleus and retrochiasmatic area as well as in the contralateral DMH. This pattern of activity is different from that produced by peripherally administered CCK which is short acting and primarily activates neurons in the nucleus of the solitary tract and area postrema, as well as the PVN and DMH. Together, these data suggest that DMH CCK plays an important role in the control of food intake, and does so by activating different pathways from those activated by peripheral CCK.

Postnatal behavioral and physiological responses of piglets from gilts housed individually or in groups during gestation.

Gestational housing of sows remains a controversial issue that may affect the well-being of both sows and piglets. Therefore, 2 types of gestational housing were used to evaluate the stress imposed on pregnant gilts by each system and the effects on the offspring by comparing production, physiology, and behavioral measures of the piglets. Forty-eight Landrace x Yorkshire gilts were randomly assigned to groups (G) of 4 per pen (n = 8 pens; 3.9 m x 2.4 m) or to individual stalls (S; n = 16 stalls; 2.21 m x 0.61 m). Gilts were moved into individual farrowing crates 5 d before the expected farrowing date. Piglets were weighed at birth, d 14, and d 35. Two barrows from each litter were weaned at d 14 (early weaning) and housed together in pens. Maintenance behaviors (head in feeder, drinking, lying, eating mash) were videotaped and observed for the first 3 d after weaning using a 10-min interval scan sampling. Belly nosing and play/fight interactions were recorded from video observations for 3 d postweaning. An isolation test (30-min duration) was performed on one piglet from each pen of barrows on d 35. Time spent lying, the number of jumps against test box walls, and grunts and squeals were recorded in real time. Salivary cortisol was collected at 30-min intervals from baseline, and 0, 30, 60, and 90 min posttest. Jugular blood was collected from 2 barrows from each litter on d 1, 7, 14, 17, 21, and 28. Plasma TNF-alpha was analyzed by ELISA, and haptoglobin, alpha1-acid glycoprotein, and immunoglobulin G were analyzed by radial immunodiffusion. More piglets from the S treatment needed to be fed a liquid feed at weaning and drank more frequently on d 2 postweaning (P < 0.05). Additionally, by d 35 piglets from S gilts had a lighter BW (10.3 kg) than G piglets (12.8 kg; P < 0.01). Piglets from S gilts also grunted more during the 30-min isolation test (number of grunts = 356) than G piglets (number of grunts = 138; P < 0.01). Salivary cortisol and immune measures were not different. These data show some behavioral and production differences between piglets from individually stalled gilts and group-housed gilts. Therefore, there may be production advantages to housing first parity gilts in groups.

Exploiting changes in the tumour microenvironment with sequential cytokine and matrix metalloprotease inhibitor treatment in a murine breast cancer model.

The study of treatment-induced changes in the tumour microenvironment might lead to effective combinations of biological therapy. IL-12 induced tumour regression and cure of an experimental murine breast cancer, HTH-K, but only after long-term treatment that was associated with chronic toxicity. During IL-12 therapy, tumour levels of the matrix metalloprotease MMP-9 declined and its inhibitor TIMP-1 was strongly induced. We therefore administered alternate cycles of IL-12 and the MMP inhibitor Batimastat (BB94) to mice. Therapeutic efficacy was increased compared with short-term IL-12 therapy but without the chronic toxicity associated with long-term IL-12 treatment. Image analysis of treated tumours revealed that BB94 prevented regeneration of tumour and stromal compartments that normally occurred after short-term IL-12 therapy.

Lack of cyclin E immunoreactivity in non-malignant breast and association with proliferation in breast cancer.

Cyclin E is a G1 cyclin that is essential for the transition from G1 to S phase in the cell cycle. Alterations to cyclin E expression or regulation could be important in tumorigenesis. Previous immunohistochemical and immunoblotting studies have investigated the expression of cyclin E in breast carcinomas. In this study, cyclin E has been investigated in a range of non-malignant and malignant breast using immunohistochemistry. Normal and benign tissue from pre- and post-menopausal women (39 cases), non-involved tissue from cancer-containing breasts (47 cases), ductal carcinoma in situ (22 cases) and invasive breast carcinomas (109 cases) have been examined. There was no reactivity in any of the non-malignant breast. Only one ductal carcinoma in situ contained more than 5% reactive cells. A total of 28% of invasive carcinomas had > 5% of reactive cells (range 0-88% positive cells, mean 12.59%, median 1.0%). A significant association was found with poorer differentiation (P < 0.001), high MIB1 index (P < 0.001), lack of oestrogen receptor (0.05 > P > 0.025) and the presence of p53 protein (0.05 > P > 0.025). Virtually all cases with cyclin E and p53 were poorly differentiated. The presence of cyclin E is therefore only found in breast malignancies and is associated with more aggressive features, including high proliferation.

Hepatic adenosine triphosphate-dependent Ca2+ transport is mediated by distinct carriers on rat basolateral and canalicular membranes.

To characterize and localize hepatic plasma membrane ATP-dependent Ca2+ transport and Na+/Ca2+ exchange, studies were performed using highly purified rat basolateral and canalicular membrane vesicles. ATP-dependent Ca2+ transport activity was present in vesicles from both domains, insensitive to azide, oligomycin, oxalate, calmodulin, and calmidazolium, and virtually abolished at pH 6.8. However, basolateral and canalicular transport differed significantly. While basolateral transport was markedly stimulated by 1 mM Mg2+, canalicular transport was Mg2+ independent. Basolateral transport was similar at pH 7.4 and 8.0 but canalicular activity was stimulated fourfold at pH 8.0. Both Ca2+ Km [1.4 +/- 0.1 (SE).10(-8) vs. 4.8 +/- 0.7.10(-8) M] and Vmax (3.6 +/- 0.1 vs. 9.0 +/- 0.6 nmol mg-1 protein min-1) were lower in basolateral than in canalicular vesicles. Basolateral transport was somewhat more nucleotide specific (for ATP) and sensitive to vanadate (IC50 130 vs. 500 microM, respectively) than was canalicular transport. Na+/Ca2+ exchange activity was not detected in membranes from either domain. These studies suggest that hepatic ATP-dependent Ca2+ transport is mediated by domain-specific carriers on the basolateral and canalicular membranes.

Estrogenic potencies of resorcylic acid lactones and 17 beta-estradiol in female rats.

Uterotrophic response in sexually immature female rats has been used to rank the relative estrogenic potencies of six resorcylic acid lactones (RALs) and to compare their activities with that of 17 beta-estradiol. On oral administration, the estrogenic potency relative to 17 beta-estradiol is as follows: 7 alpha-zearalenol, 10 times less; zeranol, 150 times less; taleranol, 350 times less; zearalanone, 400 times less; zearalenone, 650 times less; 7 beta-zearalenol, 3500 times less. On subcutaneous administration, zeranol is 500 times less estrogenic than 17 beta-estradiol.