Perinatal maternal probiotic intervention impacts immune responses and ileal mucin gene expression in a rat model of irritable bowel syndrome.

Alterations in immune responses and intestinal secretory state are among features commonly observed in the maternal separation (MS) rat model of Irritable Bowel Syndrome. This study examined whether perinatal maternal introduction of probiotics influences plasma immune markers and ileal mucin-2 (MUC2) gene expression in rat offspring exposed to neonatal maternal separation (MS, 3 h/day, postnatal days (PND) 2-14) and/or subsequently to acute restraint stress in adulthood (AS, 30 min/day, PND 83-85). Data analysis indicated that stress protocols did not affect plasma tumour necrosis factor alpha (TNF-α), interferon gamma (IFN-γ) and interleukin (IL)-6 levels in young offspring (PND 24) born to the vehicle-treated dams. Maternal probiotic intervention was associated with significantly decreased IFN-γ levels in young offspring compared with non-probiotic offspring (P≤0.05). It also induced a significant increase in IL-6 levels in MS pups (P≤0.05). Exposure of both non-MS and MS offspring to AS induced a significant increase in haptoglobin levels compared to controls (P≤0.05), whereas all offspring born to the probiotic-treated dams, irrespective of stress treatment conditions, exhibited significantly decreased haptoglobin levels to well below the control levels (P≤0.05). MS and/or AS did not affect ileal expression of MUC2 in offspring born to the non-probiotic treated dams. While maternal probiotic intake significantly downregulated ileal gene expression of MUC2 in MS male young offspring, it was associated with significantly upregulated MUC2 mRNA expression in MS or AS adult male offspring. These findings suggest that maternal probiotic intervention may exert long-lasting anti-inflammatory effects and impact gut outcomes in offspring at increased risk of dysfunctional gut.

Seasonal effects of GnIH on basal and GnRH-induced goldfish somatotrope functions.

To understand how gonadotropin-inhibitory hormone (GnIH) regulates goldfish GH cell functions, we monitored GH release and expression during early, mid-, and/or late gonadal recrudescence. In vivo and in vitro responses to goldfish (g) GnIH were different, indicating direct action at the level of pituitary, as well as interactions with other neuroendocrine factors involved in GH regulation. Injection of gGnIH consistently reduced basal serum GH levels but elevated pituitary gh mRNA levels, indicating potential dissociation of GH release and synthesis. Goldfish GnRH (sGnRH and cGnRHII) injection differentially stimulated serum GH and pituitary gh mRNA levels with some seasonal differences; these responses were reduced by gGnIH. In contrast, in vitro application of gGnIH during 24-h static incubation of goldfish pituitary cells generally elevated basal GH release and attenuated sGnRH-induced changes in gh mRNA, while suppressing basal gh mRNA levels at mid- and late recrudescence but elevating them at early recrudescence. gGnIH attenuated the GH release responses to sGnRH during static incubation at early, but not at mid- and late recrudescence. In cell column perifusion experiments examining short-term GH release, gGnIH reduced the cGnRHII- and sGnRH-stimulated secretion at late recrudescence but inhibited tha action of cGnRHII only during mid-recrudescence. Interestingly, a reduction of basal GH release upon perifusion with gGnIH during late recrudescence was followed by a rebound increase in GH release upon gGnIH removal. These results indicate that gGnIH exerts complex effects on basal and GnRH-stimulated goldfish GH cell functions and can differentially affect GH release and mRNA expression in a seasonal reproductive manner.

Seasonal effect of gonadotrophin inhibitory hormone on gonadotrophin-releasing hormone-induced gonadotroph functions in the goldfish pituitary.

We have shown that native goldfish gonadotrophin inhibitory hormone (gGnIH) differentially regulates luteinsing hormone (LH)-ß and follicle-stimulating hormone (FSH)-ß expression. To further understand the functions of gGnIH, we examined its interactions with two native goldfish gonadotrophin-releasing hormones, salmon gonadotrophin-releasing hormone (sGnRH) and chicken (c)GnRH-II in vivo and in vitro. Intraperitoneal injections of gGnIH alone reduced serum LH levels in fish in early and mid gonadal recrudescence; this inhibition was also seen in fish co-injected with either sGnRH or cGnRH-II during early recrudescence. Injection of gGnIH alone elevated pituitary LH-ß and FSH-ß mRNA levels at early and mid recrudescence, and FSH-ß mRNA at late recrudescence. Co-injection of gGnIH attenuated the stimulatory influences of sGnRH on LH-ß in early recrudescence, and LH-ß and FSH-ß mRNA levels in mid and late recrudescence, as well as the cGnRH-II-elicited increase in LH-ß, but not FSH-ß, mRNA expression at mid and late recrudescence. sGnRH and cGnRH-II injection increased pituitary gGnIH-R mRNA expression in mid and late recrudescence but gGnIH reduced gGnIH-R mRNA levels in late recrudescence. gGnIH did not affect basal LH release from perifused pituitary cells and continual exposure to gGnIH did not alter the LH responses to acute applications of GnRH. However, a short 5-min GnIH treatment in the middle of a 60-min GnRH perifusion selectively reduced the cGnRH-II-induced release of LH. These novel results indicate that, in goldfish, gGnIH and GnRH modulate pituitary GnIH-R expression and gGnIH differentially affects sGnRH and cGnRH-II regulation of LH secretion and gonadotrophin subunit mRNA levels. Furthermore, these actions are manifested in a reproductive stage-dependent manner.

Fibroblast growth factor and ornithine decarboxylase 5'UTRs enable preferential expression in human prostate cancer cells and in prostate tumors of PTEN(-/-) transgenic mice.

In this study, we have taken advantage of over-expression of eukaryotic translation initiation factor 4E (eIF4E) in prostate cancer cells to design a viral-based targeting system of prostate cancer. Three different lengths of 5'-untranslated regions (5'UTRs) derived from either fibroblast growth factor-2 (FU-FGF2-GW) or ornithine decarboxylase (FU-ODC149-GW and FU-ODC274-GW) were inserted upstream of enhanced green fluorescent protein (GFP) gene in a lentiviral backbone. Both nonmalignant control (PNT1B and BPH-1) and neoplastic (LNCaP, C4-2, DU145 and PC-3) prostate cell lines were transfected with each plasmid or virus alone, or in the presence of siRNA against eIF4E, and their expression was monitored via GFP protein levels. Two 5'UTRs (FU-FGF2-GW and FU-ODC-GW) were selected as being most sensitive to eIF4E status. Lentiviruses containing these sequences were injected directly into the prostates of PTEN(-/-) (tumor-bearing) and control mice. Immunofluorescence data and western blot analyses determined that a lentivirus containing a 5'UTR derived from FGF-2 is the best candidate for directing selective gene expression in the prostate tumors of PTEN(-/-) mice in vivo. This study demonstrates that judicious selection of a complex 5'UTR can enhance selective targeting of viral-based gene therapies for prostate cancer.

Seasonal effect of GnIH on gonadotrope functions in the pituitary of goldfish.

Gonadotropin-inhibitory hormone (GnIH) inhibits gonadotropin release in birds and mammals. To investigate its role in teleosts, we examined the effects of synthetic goldfish (g)GnIH on pituitary LH-ß and FSH-ß subunit, and gGnIH receptor (gGnIH-R) mRNA levels and LH secretion in goldfish. Intraperitoneal injections of gGnIH increased pituitary LH-ß and FSH-ß mRNA levels at early to late gonadal recrudescence, but reduced serum LH and pituitary gGnIH-R mRNA levels, respectively, at early to mid-recrudescence and later stages of recrudescence. Static incubation with gGnIH elevated LH secretion from dispersed pituitary cell cultures from prespawning fish, but not at other recrudescent stages; suppressed LH-ß mRNA levels at early recrudescence and prespawning but elevated LH-ß at mid-recrudescence; and consistently attenuated FSH-ß mRNA in a dose-specific manner. Results indicate that in goldfish, regulation of LH secretion and gonadotropin subunit mRNA levels are dissociated in the presence of gGnIH and dependent on maturational status and administration route.

Lentiviruses with trastuzumab bound to their envelopes can target and kill prostate cancer cells.

In this study, we took advantage of the overexpression of human epidermal growth factor receptor 2 (HER-2) in prostate cancers to design lentiviruses with modified envelope proteins that bind antibodies to specific cell-surface antigens. When bound to trastuzumab (Herceptin, Genentech, CA), lentiviruses were able to selectively infect androgen-sensitive LNCaP and castration-resistant C4-2 human prostate cancer cell lines, both of which express high levels of HER-2. To test for a therapeutic effect, we engineered our antibody-binding lentiviruses to express thymidine kinase, which can convert the non-toxic pro-drug ganciclovir (GCV) into a cytotoxic form. LNCaP and C4-2 cells infected by these viruses were sensitive to GCV killing. In vivo, C4-2 xenograft tumors treated either intratumorally or i.v. with trastuzumab-bound lentivirus expressed luciferase, although the latter route was less tumor specific. When a prostate-specific promoter for governing luciferase expression was combined with trastuzumab-mediated delivery, there was a further enrichment in targeting viral gene expression in prostate tumors. In conclusion, we found that although prostate cancers that express high levels of HER-2 are resistant to the killing effects of trastuzumab, they can be targeted for selective gene expression and destruction by viruses with envelope proteins engineered to bind this antibody.

Chemical pretreatment of formaldehyde-containing effluents.

Lime was found in this study to be an efficient reagent to lower the concentration of formaldehyde in highly concentrated effluents down to and below the limits suitable for biological treatment systems. The results show that the reactions leading to formaldehyde elimination can be divided in two steps. In the first step, the reaction is relatively slow. More than two-thirds of the original formaldehyde disappears in the second step in a period as short as one-third of the first step. Such trend is followed in a temperature range of up to 92 degrees C. Economical considerations suggest maintaining the conditions of the process around the ambient temperature with no heat requirement. It was noticed that the efficiencies of formaldehyde removal better than 99% could be achievable even around room temperature. However, these efficiencies would result in quite a shorter period of time if the temperature was raised. The mathematical representation for the rate of formaldehyde removal was found to appear with an exponential behavior. It will be seen that the rate of formaldehyde removal is strongly dependent on temperature. The present survey proves that the formaldehyde-containing effluents can be treated in a pretreatment step by lime to maintain the formaldehyde concentration in a range that is safe for biological treatment systems.