Anticancer actions of carnosine in cellular models of prostate cancer.

Treatments for organ-confined prostate cancer include external beam radiation therapy, radical prostatectomy, radiotherapy/brachytherapy, cryoablation and high-intensity focused ultrasound. None of these are cancer-specific and are commonly accompanied by side effects, including urinary incontinence and erectile dysfunction. Moreover, subsequent surgical treatments following biochemical recurrence after these interventions are either limited or affected by the scarring present in the surrounding tissue. Carnosine (ß-alanyl-L-histidine) is a histidine-containing naturally occurring dipeptide which has been shown to have an anti-tumorigenic role without any detrimental effect on healthy cells; however, its effect on prostate cancer cells has never been investigated. In this study, we investigated the effect of carnosine on cell proliferation and metabolism in both a primary cultured androgen-resistant human prostate cancer cell line, PC346Flu1 and murine TRAMP-C1 cells. Our results show that carnosine has a significant dose-dependent inhibitory effect in vitro on the proliferation of both human (PC346Flu1) and murine (TRAMP-C1) prostate cancer cells, which was confirmed in 3D-models of the same cells. Carnosine was also shown to decrease adenosine triphosphate content and reactive species which might have been caused in part by the increase in SIRT3 also shown after carnosine treatment. These encouraging results support the need for further human in vivo work to determine the potential use of carnosine, either alone or, most likely, as an adjunct therapy to surgical or other conventional treatments.

Longitudinal Changes in Cerebral Perfusion, Perivascular Space Volume, and Ventricular Volume in a Healthy Cohort Undergoing a Spaceflight Analog.

BACKGROUND AND PURPOSE: A global decrease in brain perfusion has recently been reported during exposure to a ground-based spaceflight analog. Considering that CSF and glymphatic flow are hypothesized to be propelled by arterial pulsations, it is unknown whether a change in perfusion would impact these CSF compartments. The aim of the current study was to evaluate the relationship among changes in cerebral perfusion, ventricular volume, and perivascular space volume before, during, and after a spaceflight analog. MATERIALS AND METHODS: Eleven healthy participants underwent 30 days of bed rest at 6° head-down tilt with 0.5% atmospheric CO2 as a spaceflight analog. For each participant, 6 MR imaging brain scans, including perfusion and anatomic-weighted T1 sequences, were obtained before, during, and after the analog period. Global perfusion, ventricular volume, and perivascular space volume time courses were constructed and evaluated with repeated measures ANOVAs. RESULTS: Global perfusion followed a divergent time trajectory from ventricular and perivascular space volume, with perfusion decreasing during the analog, whereas ventricular and perivascular space volume increased (P < .001). These patterns subsequently reversed during the 2-week recovery period. CONCLUSIONS: The patterns of change in brain physiology observed in healthy participants suggest a relationship between cerebral perfusion and CSF homeostasis. Further study is warranted to determine whether a causal relationship exists and whether similar neurophysiologic responses occur during spaceflight.

Circular dichroism in biological photonic crystals and cubic chiral nets.

Nature provides impressive examples of chiral photonic crystals, with the notable example of the cubic so-called srs network (the label for the chiral degree-three network modeled on SrSi2) or gyroid structure realized in wing scales of several butterfly species. By a circular polarization analysis of the band structure of such networks, we demonstrate strong circular dichroism effects: The butterfly srs microstructure, of cubic I4(1)32 symmetry, shows significant circular dichroism for blue to ultraviolet light, that warrants a search for biological receptors sensitive to circular polarization. A derived synthetic structure based on four like-handed silicon srs nets exhibits a large circular polarization stop band of a width exceeding 30%. These findings offer design principles for chiral photonic devices.

Proteins are secreted by both constitutive and regulated secretory pathways in lactating mouse mammary epithelial cells.

Lactating mammary epithelial cells secrete high levels of caseins and other milk proteins. The extent to which protein secretion from these cells occurs in a regulated fashion was examined in experiments on secretory acini isolated from the mammary glands of lactating mice at 10 d postpartum. Protein synthesis and secretion were assayed by following the incorporation or release, respectively, of [35S]methionine-labeled TCA-precipitable protein. The isolated cells incorporated [35S]methionine into protein linearly for at least 5 h with no discernible lag period. In contrast, protein secretion was only detectable after a lag of approximately 1 h, consistent with exocytotic secretion of proteins immediately after passage through the secretory pathway and package into secretory vesicles. The extent of protein secretion was unaffected by the phorbol ester PMA, 8-bromo-cAMP, or 8-bromo-cGMP but was doubled by the Ca2+ ionophore ionomycin. In a pulse-label protocol in which proteins were prelabeled for 1 h before a chase period, constitutive secretion was unaffected by depletion of cytosolic Ca2+ but ionomycin was found to give a twofold stimulation of the secretion of presynthesized protein in a Ca(2+)-dependent manner. Ionomycin was still able to stimulate protein secretion after constitutive secretion had terminated. These results suggest that lactating mammary cells possess both a Ca(2+)-independent constitutive pathway and a Ca(2+)-activated regulatory pathway for protein secretion. The same proteins were secreted by both pathways. No ultrastructural evidence for apocrine secretion was seen in response to ionomycin and so it appears that regulated casein release involves exocytosis. Ionomycin was unlikely to be acting by disassembling the cortical actin network since cytochalasin D did not mimic its effects on secretion. The regulated pathway may be controlled by Ca2+ acting at a late step such as exocytotic membrane fusion.

A novel TNFRSF1A splice mutation associated with increased nuclear factor kappaB (NF-kappaB) transcription factor activation in patients with tumour necrosis factor receptor associated periodic syndrome (TRAPS).

OBJECTIVE: To characterise and investigate the functional consequences of a novel TNFRSF1A splice site mutation causing tumour necrosis factor receptor associated periodic syndrome (TRAPS) in a 16-year-old male patient and his mother. METHODS: Mutational DNA screening was performed in the patient and his mother. Western blotting was used to analyse protein expression levels of TNFR1. A multiplex bead immunoassay was used to quantify serum levels of range of cytokines, and an ELISA-based transcription factor assay to measure nuclear factor (NF)-kappaB transactivation. Serum levels of soluble TNFR1 (sTNFR1) were measured by ELISA and fluorescence-activated cell sorting (FACS) analysis used to measure monocyte TNFR1 cell surface expression. RESULTS: A novel mutation, c.472+1G>A (C158delinsYERSSPEAKPSPHPRG), involving a splice site in intron 4 of TNFRSF1A, was found in the proband and affected mother leading to a 45 nucleotide insertion of intronic DNA into the mRNA, resulting in an in-frame insertion of 15 amino acids in the mature TNFR1 protein and a deletion of a cysteine residue C129 (158) in cysteine rich domain (CRD)3. The patients had reduced serum sTNFR1 and surface expression levels of TNFR1, with marked increases in pro- and anti-inflammatory cytokine. Their peripheral blood mononuclear cells (PBMC) had increased basal NF-kappaB activation compared with healthy controls and also had increased p50 nuclear expression following tumour necrosis factor (TNF) stimulation compared with PBMC from healthy controls, as well as T50M (T79M) and C88R (C117R) patients with TRAPS and patients with rheumatoid arthritis (RA). CONCLUSION: A novel, TRAPS causing, TNFRSF1A splice site mutation is associated with decreased sTNFR1 levels, cell surface and whole cell extract expression and increased NF-kappaB transcription factor activation.

Groundtruthing and potential for predicting acid deposition impacts in headwater streams using bedrock geology, GIS, angling, and stream chemistry.

Atmospheric acid deposition is of environmental concern worldwide, and the determination of impacts in remote areas can be problematic. Rainwater in central Pennsylvania, USA, has a mean pH of approximately 4.4. Bedrock varies dramatically in its ability to neutralize acidity. A GIS database simplified reconnaissance of non-carbonate bedrock streams in the Valley and Ridge Province and identified potentially chronically impacted headwater streams, which were sampled for chemistry and brook trout. Stream sites (n=26) that originate in and flow through the Tuscarora had a median pH of 5.0 that was significantly different from other formations. Shawangunk streams (n=6) and non-Tuscarora streams (n=20) had a median pH of 6.0 and 6.3, respectively. Mean alkalinity for non-Tuscarora streams (2.6 mg/L CaCO(3)) was higher than the mean for Tuscarora streams (0.5 mg/L). Lower pH and alkalinity suggest that the buffering capability of the Tuscarora is inferior to that of adjacent sandstones. Dissolved aluminum concentrations were much higher for Tuscarora streams (0.2 mg/L; approximately the lethal limit for brook trout) than for non-Tuscarora streams (0.03 mg/L) or Shawangunk streams (0.02 mg/L). Hook-and-line methods determined the presence/absence of brook trout in 47 stream reaches with suitable habitat. Brook trout were observed in 21 of 22 non-Tuscarora streams, all 6 Shawangunk streams, and only 9 of 28 Tuscarora stream sites. Carefully-designed hook-and-line sampling can determine the presence or absence of brook trout and help confirm biological impacts of acid deposition. 15% of 334 km of Tuscarora stream lengths are listed as "impaired" due to atmospheric deposition by the Pennsylvania Department of Environmental Protection. 65% of the 101 km of Tuscarora stream lengths examined in this study were impaired.

"COLO 357," a human pancreatic adenosquamous carcinoma: growth in artificial capillary culture and in nude mice.

The human pancreatic cancer cell line COLO 357 has been xenografted s.c. in athymic Swiss mice. The xenografts grew well to form typical adenosquamous carcinomas. The cells were placed in a perfused artificial capillary system where they formed a solid tumor mass which survived for 7 weeks. In this system, the cells consumed glucose and released enzymes and carcinoembryonic antigen into the extracapillary space.

Establishment and characterization of two human pancreatic cancer cell lines tumorigenic in athymic mice.

Two human pancreatic cancer lines, RWP-1 and RWP-2, have been established from 2 patients with primary pancreatic cancer metastatic to the liver. The patients' tumors, the xenografted tumors, and tumors obtained by inoculation of nude mice with cultured RWP-1 and RWP-2 cells are all moderately-well-differentiated ductal cell adenocarcinomas. Ultrastructural analysis supports the tissue histopathology findings. Xenografts of RWP-1 tumors double every 10 days, whereas the doubling time of RWP-2 xenografts is 22 days. Both tumors contain mucin. RWP-1 and RWP-2 cells have a doubling time in culture of 45 hr and form colonies in soft agar. RWP-1 cultures appear to be morphologically heterogeneous; two distinct epithelial cell types can be identified. RWP-1 and RWP-2 have modal chromosome numbers of 64 and 62, respectively. Appreciable levels of glucose-6-phosphate dehydrogenase and lactic dehydrogenase were found in both cell lines and xenografts. RWP-1 and RWP-2 cells produce appreciable amounts of carcinoembryonic antigen, 1090 and 414 ng/10(6) cells, respectively.

The Effect of a Combination Saliva Substitute for the Management of Xerostomia and Hyposalivation.

OBJECTIVE: The objective of this study was to evaluate the difference between the combination agent of xylitol, beatine and olive oil in a chewable capsule versus the control agent of a sorbitol tablet in subjects with hyposalivation and xerostomia. MATERIALS AND METHODS: The subjects had xerostomia over 3 months and a measured hyposalivation. The study was 3 weeks in duration, with 2 treatment phases of 1 week and a 7 day wash out period in between. At the end of each treatment phase, subjects returned for a follow up evaluation. At this visit they were given the subjective sensation questionnaire, as well as their unstimulated whole salivary flow and stimulated whole salivary flow were measured. RESULTS: There was a greater increase in the unstimulated and stimulated whole salivary flow rate, although the results were not statistically significant. The subjective evaluation as measured by the questionnaire showed that both agents reduced the mean score as compared to the baseline, although only the findings in the active agent was statistically significant (p = 0.0015). CONCLUSION: The significant conclusions found in this study were that the active agent provided a significant subjective improvement in speech, swallowing, and decreased subjective xerostomia as compared to the control tablet. CLINICAL RELEVANCE: This combination agent has a significant effect on patients with subjective xerostomia but does not have a significant effect on objective hyposalivation.

Experimental small bowel transplantation utilizing cyclosporine.

Cyclosporine prolonged the survival of ectopic small bowel allografts in a canine model. A 9-fold increase in mean survival as compared with controls was obtained. Addition of prednisone to cyclosporine did not result in further graft prolongation, but improved the gross and histological appearance of the allograft. Monitoring the motility and histology of the allograft appears to be useful in predicting approaching rejection. Xylose absorption was not helpful in this regard. A new technique for measuring fat absorption in the transplant is described. Following allografting, fat absorption is resumed by 14 days posttransplantation.

Radiological evaluation of rat colonic tumors.

Colon carcinomata were induced in rats with N-methyl-N-nitro-N-nitrosoguanidine. A magnified double contrast barium enema technique was developed for an in vivo study of tumor growth patterns. Invariably, all small tumors were smoothly outlined and were spherical or ellipsoidal in shape. Irregularity developed only when the tumors grew larger, at which time there was considerable variation in the tumor growth patterns. The onset, shape, progression, spread and number of tumors present can be studied without sacrificing the animal.

Successful orthotopic allotransplantation of hearts from dogs in irreversible shock.

The viability of the heart from irreversibly shocked dogs was evaluated. Thirty-one mongrel dogs were used to develop a uniformly lethal hemorrhagic shock model. The final model utilized a 30 mm. Hg mean arterial pressure with a 40 percent decompensation (uptake of reservoir blood). All control dogs subjected to this procedure died within 24 hours. Eleven hearts were transplanted following the shock procedure. Blood samples were drawn at intervals and analyzed for Po2, pH, glucose, oxygen content, and hematocrit. Cardiac outputs and ECG's were taken before and after transplant. All parameters measured returned to control levels following the transplants. Of the dogs receiving heart transplants, 10 survived more than 24 hours and eight survived more than 7 days. Compared with the survival of the shock model experiments, the survival of the transplant recipients was significant at the p less than 0.01 level. These results suggest that deterioration of the heart is not the factor which prevents recovery from irreversible hemorrhagic shock.

A rabbit tooth-pulp assay to quantify efficacy and duration of antinociception by local anesthetics infiltrated into maxillary tissues.

The rabbit tooth-pulp assay is well established as a method for measuring the efficacy and potency of parenteral analgesic drugs. We describe a method for administration of local anesthetic drugs into the maxillary arch and subsequent measurements of antinociceptive action. It was possible to use two different methods of ED50 estimation and to provide measures of the potency, efficacy, and duration of local anesthetic drugs. These measurements corresponded with in vitro estimates of potency and duration and with intrinsic observations of the clinical actions.

Fetal methylmercury study in a Peruvian fish-eating population.

Maternal consumption during pregnancy of methylmercury (MeHg)-contaminated fish in Japan and of MeHg-contaminated bread in Iraq caused psychomotor retardation in the offspring. Studies in Iraq suggested adverse fetal effects when maternal hair mercury concentrations were as low as 20 ppm. This prospective study involved 131 infant-mother pairs in Mancora, Peru with peak maternal hair MeHg levels during pregnancy from 1.2 ppm to 30.0 ppm, geometric mean 8.3. The MeHg was believed to be derived from marine fish in the diet. There was no increase in the frequency of neurodevelopmental abnormalities in early childhood. The possible role of selenium or other protective mechanisms in marine fish is discussed. This previously unpublished study was conducted between 1981 and 1984. Our report of August 1985 to the funding agencies has been circulated, and the data were presented at the Twelfth International Neurotoxicology Conference in Hot Spring, Arkansas, October 30 to November 2, 1994. The current account has not been modified or updated since 1985. For reference to interim publications on fetal MeHg studies in Iraq and New Zealand see Marsh et al., 1995.

The effects of nicotine on learning and memory: a neuropsychological assessment in young and senescent Fischer 344 rats.

The effects of chronic nicotine on the behavioral performance of young (4 month) and old (24 month) Fischer-344 rats were assessed on four behavioral tasks: activity chamber. rotating rod, serial pattern learning, and Morris water maze paradigm. Old and young nicotine-treated rats received an intraperitoneal injection of nicotine (0.20 mg/kg) 15 min prior to all behavioral testing, and old and young saline-treated rats received saline injections 15 min prior to all behavioral testing. Nicotine improved motor coordination and increased the general activity levels of the old rats compared to old saline-treated rats. There were no significant differences in the behaviors of the young rats in these behavioral evaluations. In young rats, nicotine improved the acquisition of a serial pattern, suggesting an improvement in working memory or related processes. Nicotine was found to increase swim speed in a Morris water maze paradigm with a hidden platform; however, no beneficial effects of nicotine in reference memory were obtained for either age group. These results suggests that nicotine may not be as beneficial in attenuating age-related learning and memory deficits as once proposed.

Hemorrhagic shock in dogs. Comparison of treatment with shed blood alone versus shed blood plus Ringer's lactate: intravascular pressures, cardiac output, oxygen consumption, arteriovenous oxygen differences, extracellular fluid PO2, electrolyte changes, and survival rates.

The purpose of our study of hemorrhagic shock in dogs was to examine the efficacy of adding Ringer's lactate to shed blood replacement in increasing animal rates. The standard Wiggers' hemorrhagic shock technic was used in two groups of ten animals each. Intravascular pressures, cardiac outut, oxygen consumption, arteriovenous oxygen differences, extracellular fluid PO2 and pH, electrolyte changes, and survival rates were determined. There was a positive correlation between changes in cardiac output, central venous oxygen content, and PO2 and extracellular fluid PO2 as measured using subcutaneously implanted Silastic tubing and perforated plastic balls. Four of the dogs that received shed blood alone survived, whereas five of the dogs that received shed blood plus Ringer's lactate survived. This difference was not statistically significant.

Methylmercury in populations eating large quantities of marine fish.

A Peruvian population was identified that was chronically exposed to methylmercury from the long-term consumption of ocean fish. The weekly fish intake averaged 10.1 kg per average family of 6.2 persons. Blood methylmercury concentrations ranged from 11 to 275 ng/ml, with a mean of 82 ng/ml. Paresthesias were reported by 29.5% of the population. In contrast, a nearby control population had a mean weekly fish consumption of 1.9 kg per average family of 6.4 persons. Their blood methylmercury levels were 3.3-25.1 ng/ml, with a mean of 9.9 ng/ml. Paresthesias were reported by 49.5% of this control group. No individual was identified with symptoms or signs that could be attributed to methylmercury intoxication.