Differential Calcium Dependence of Axonal Versus Somatodendritic Dopamine Release, with Characteristics of Both in the Ventral Tegmental Area

Midbrain dopamine (DA) neurons in the substantia nigra pars compacta (SNc) and ventral tegmental area (VTA) exhibit somatodendritic release of DA. Previous studies indicate a difference between the Ca2+ dependence of somatodendritic DA release in the SNc and that of axonal DA release in dorsal striatum. Here, we evaluated the Ca2+ dependence of DA release in the VTA and nucleus accumbens (NAc) shell for comparison with that in the SNc and dorsal striatum. Release of DA was elicited by single-pulse stimulation in guinea-pig brain slices and monitored with subsecond resolution using carbon-fiber microelectrodes and fast-scan cyclic voltammetry. In dorsal striatum and NAc, DA release was not detectable at extracellular Ca2+ concentrations ([Ca2+]o) below 1 mM; however, a progressive increase in evoked extracellular DA concentration ([DA]o) was seen with [Ca2+]o ≥ 1.5 mM. By contrast, in SNc and VTA, robust increases in [DA]o could be elicited in 0.25 mM [Ca2+]o that were ∼60% of those seen in 1.5 mM [Ca2+]o. In SNc, a plateau in single-pulse evoked [DA]o was seen at [Ca2+]o ≥ 1.5 mM, mirroring the release plateau reported previously for pulse-train stimulation in SNc. In VTA, however, evoked [DA]o increased progressively throughout the range of [Ca2+]o tested (up to 3.0 mM). These functional data are consistent with the microanatomy of the VTA, which includes DA axon collaterals as well as DA somata and dendrites. Differences between axonal and somatodendritic release data were quantified using Hill analysis, which showed that the Ca2+ dependence of axonal DA release is low affinity with high Ca2+ cooperativity, whereas somatodendritic release is high affinity with low cooperativity. Moreover, this analysis revealed the dual nature of DA release in the VTA, with both somatodendritic and axonal contributions

α2β1 integrin affects metastatic potential of ovarian carcinoma spheroids by supporting disaggregation and proteolysis

Background: Ovarian cancer is characterized by a wide-spread intra-abdominal metastases which represents a major clinical hurdle in the prognosis and management of the disease. A significant proportion of ovarian cancer cells in peritoneal ascites exist as multicellular aggregates or spheroids. We hypothesize that these cellular aggregates or spheroids are invasive with the capacity to survive and implant on the peritoneal surface. This study was designed to elucidate early inherent mechanism(s) of spheroid survival, growth and disaggregation required for peritoneal metastases.Methods: In this study, we determined the growth pattern and adhesive capacity of ovarian cancer cell lines (HEY and OVHS1) grown as spheroids, using the well established liquid overlay technique, and compared them to a normal ovarian cell line (IOSE29) and cancer cells grown as a monolayer. The proteolytic capacity of these spheroids was compared with cells grown as a monolayer using a gelatin zymography assay to analyze secreted MMP-2/9 in conditioned serum-free medium. The disaggregation of cancer cell line spheroids was determined on extracellular matrices (ECM) such as laminin (LM), fibronectin (FN) and collagen (CI) and the expression of &alpha;2, &alpha;3, &alpha;v, &alpha;6 and &beta;1 interin was determined by flow cytometric analysis. Neutralizing antibodies against &alpha;2, &beta;1 subunits and &alpha;2&beta;1 integrin was used to inhibit disaggregation as well as activation of MMPs in spheroids.Results: We demonstrate that ovarian cancer cell lines grown as spheroids can sustain growth for 10 days while the normal ovarian cell line failed to grow beyond 2 days. Compared to cells grown as a monolayer, cancer cells grown as spheroids demonstrated no change in adhesion for up to 4 days, while IOSE29 cells had a 2&ndash;4-fold loss of adhesion within 2 days. Cancer cell spheroids disaggregated on extracellular matrices (ECM) and demonstrated enhanced expression of secreted pro-MMP2 as well as activated MMP2/MMP9 with no such activation of MMP\u27s observed in monolayer cells. Flow cytometric analysis demonstrated enhanced expression of &alpha;2 and diminution of &alpha;6 integrin subunits in spheroidsversus monolayer cells. No change in the expression of &alpha;3, &alpha;v and &beta;1 subunits was evident. Conversely, except for &alpha;v integrin, a 1.5&ndash;7.5-fold decrease in &alpha;2, &alpha;3, &alpha;6 and &beta;1 integrin subunit expression was observed in IOSE29 cells within 2 days. Neutralizing antibodies against &alpha;2, &beta;1 subunits and &alpha;2&beta;1 integrin inhibited disaggregation as well as activation ofMMPs in spheroids.Conclusion: Our results suggest that enhanced expression of &alpha;2&beta;1 integrin may influence spheroid disaggregation andproteolysis responsible for the peritoneal dissemination of ovarian carcinoma. This may indicate a new therapeutic targetfor the suppression of the peritoneal metastasis associated with advanced ovarian carcinomas.<br /

Nurses\u27 Alumnae Association Bulletin - Volume 5 Number 8

Calling All Nurses Financial Report Calendar of Events Lest You Forget! Attention Review of the Alumnae Association Meetings President\u27s Report Barton Memorial Division Oxygen Therapy Welcome, White Haven Alumnae Clinical Use of Penicillin in Infections of the Ears, Nose and Throat Address - Graduation of Nurses, 1945 Miscellaneous Items The Blood that Kills The Story of Malaria Program Prizes - May, 1946 Capping Exercises The Economic Security Program of the Pennsylvania State Nurses\u27 Association The Clara Melville Scholarship Fund Card of Thanks The Poet\u27s Corner The Hospital Pharmacy Jefferson Medical College Hospital School of Nursing Faculty Jefferson Hospital Gray Lady Unite, A.R.R. The Volunteer Nurses\u27 Aides Salute Jefferson Nurses Changes in the Staff at Jefferson Hospital Red Cross Recruits Did You Know That The Pennsylvania Nurse Medical College News Magazine and Newspaper Items Central Dressing Room and Transfusion Unit Rules Concerning Central Dressing Room Radios and Electrical Appliances Attending College Nurses in Anesthesia Condolences Marriages New Arrivals Deaths The Bulletin Committee Attention, Alumnae New Addresse

Overexpression of Strigolactone-Associated Genes Exerts Fine-Tuning Selection on Soybean Rhizosphere Bacterial and Fungal Microbiome

Strigolactones are a recently discovered class of carotenoid-derived plant hormones with a wide variety of functions, including acting as signaling molecules in the rhizosphere to promote arbuscular mycorrhizal fungi colonization and parasitic seed germination. To determine whether strigolactones influence the recruitment of microbes to the rhizosphere, we characterized both bacterial and fungal communities in response to the overexpression of genes involved in strigolactone biosynthesis (MAX1d) and signaling perception (D14 and MAX2a) in soybean (Glycine max). The amplicon sequencing-based results suggest that strigolactone overexpression lines had altered soybean rhizosphere bacteria composition at both the community level and individual taxa level with genera including Shinella and Bdellovibrio consistently more abundant across all three overexpression constructs. In addition, the responses of the soybean rhizosphere bacteria community differed significantly across the genes, with lines overexpressing genes involved in strigolactone biosynthesis (MAX1d) yielding a divergent bacterial community in comparison with those with altered expression of strigolactone perception genes (D14 and MAX2a). The overexpressed genes did not significantly impact the overall fungal community distribution; however, some individual taxa were altered in abundance. MAX1d and D14 overexpression lines had significantly enriched abundance of Fusarium solani. The mediating role of strigolactone biosynthesis and signaling genes on soybean rhizosphere bacterial and fungal communities confirmed strigolactone’s importance in the rhizosphere host–microbe communication and microbial community structure

Psilocybin prevents symptoms of hyperarousal and enhances novel object recognition in rats exposed to the single prolonged stress paradigm

Pharmacotherapy for stress-related psychological disorders remains inadequate. Patients who are treated with conventional pharmacological agents frequently report negligeable symptom reduction, and, in most cases, less than 50% experience full remission. Clearly, there is a need for additional pharmaceutical research into both established and novel approaches to alleviate these conditions. Over the past several years, there has been a renewed interest in the use of psychedelics to aid in the treatment of psychological disorders. Several studies have reported promising results in patients with major depression, anxiety disorders, and post-traumatic stress disorder (PTSD) following treatment with psychedelic agents such as lysergic acid diethylamide (LSD), 3,4-methylenedioxymethamphetamine (MDMA), ayahuasca, ketamine, and psilocybin. However, the precise behavioral and neurobiological mechanisms for these effects remain unclear. Thus, we aimed to develop an animal model of PTSD that involved prophylactic treatment with psilocybin, a 5-HT2A agonist, that could be used to further understand the mechanisms underlying the benefit of psychedelic substances in treating these disorders. Adult male and female Sprague-Dawley rats were subjected to the single prolonged stress (SPS) paradigm, including 2 hours of physical restraint, 15 minutes of forced swim, and ether vapor exposure until loss of consciousness. Five minutes following ether-induced loss of consciousness, the rats were intraperitoneally injected with vehicle (0.9% saline) or psilocybin (1 mg/kg). One week later, the rats underwent a battery of behavioral tests, including the elevated plus maze (EPM), startle response assessment, open field testing, and novel object recognition (NOR) testing. No effects of SPS or psilocybin were observed for EPM behavior. SPS led to enhanced startle responses in males, but not females, which was prevented by psilocybin. SPS also increased locomotor activity in the open field in males, but not females, and this effect was not prevented by psilocybin. SPS had no impact on NOR memory in males, but enhanced memory in females. Interestingly, psilocybin administration, alone or in combination with SPS, enhanced NOR memory in males only. These findings support a complex interaction between the administration of psilocybin and the prevention of stress-induced behavioral sequelae that depends on both sex and the type of behavioral task

Low-dose psilocybin enhances novel object recognition but not inhibitory avoidance in adult rats

Given the recently renewed interest in using psychedelics to aid in the treatment of psychological disorders, we aimed to examine the impact of psilocybin, a 5-HT2A agonist, on learning and memory in rodents. Previous work has demonstrated that psilocybin and other 5-HT2A agonists can enhance fear conditioning, fear extinction, and novel object recognition (NOR). Thus, we predicted that low doses of psilocybin would enhance inhibitory avoidance (IA) and NOR memory. In the first experiment, adult male and female Sprague-Dawley rats underwent step-through IA training (involving 0.45, 0.65, or 1 mA scrambled footshock) and were injected intraperitoneally (i.p.) with vehicle (0.9% saline) or psilocybin (1 mg/kg) immediately afterward. Rats were tested for their IA memory two days later. In the second experiment, adult male and female Sprague-Dawley rats were acclimated to an open field apparatus for 5 minutes on Day 1. The next day, the rats were given i.p. injections of vehicle or psilocybin (0.1 mg/kg) 10 minutes before undergoing NOR training, during which they were exposed to two replicas of an identical object for 3 minutes. On Day 3, one of the objects from NOR training was exchanged for a novel object; rats were exposed to this novel object and a new replica of the object from Day 2 (i.e., familiar object) for 5 minutes. The results showed that psilocybin had no significant impact on IA memory but enhanced novel object recognition memory in both males and females. The differential impact of psilocybin on IA memory and novel object recognition could be explained by the different doses of psilocybin or the different times of drug administration used for each task. Alternatively, they may suggest that psilocybin exerts distinct effects on different types of learning

Reproductive factors and hormone use and risk of adult gliomas

Previous research suggests there may be a hormonal influence on glioma risk as evidenced by lower rates in females, change in incidence rates around ages at menarche and menopause and presence of hormone receptors in glial tumors. Using the large San Francisco Bay Area Adult Glioma Study, we investigated whether reported reproductive factors and hormone use were associated with gliomas overall or with histologic subtypes among female cases (n=619) and controls (n=650). We found that reproductive factors were generally not associated with gliomas. Weak to moderately elevated odds ratios were observed for self-reported later age at menarche (14+ years old versus 12–13 years old: adjusted odds ratio (AOR) = 1.39, 95% confidence interval (CI): 1.02 –1.89), particularly for non-glioblastoma histologies (AOR = 1.64, 95% CI: 1.11–2.43). Inverse associations were observed for ever self-reported use of exogenous hormones (oral contraceptive use: AOR = 0.72, 95% CI: 0.53–0.99; postmenopausal hormone use: AOR = 0.56, CI: 0.37–0.84). However, cumulative hormone exposure defined multiple ways demonstrated no clear pattern of association. The results of this study suggest that any protective effect of hormones on gliomas may be limited to exogenous hormones, but a more detailed history of exogenous hormone use are needed to confirm findings

Differential Regulation of Lipoprotein and Hepatitis C Virus Secretion by Rab1b

Secretory cells produce diverse cargoes, yet how they regulate concomitant secretory traffic remains insufficiently explored. Rab GTPases control intracellular vesicular transport. To map secretion pathways, we generated a library of lentivirus-expressed dominant-negative Rab mutants and used it in a large-scale screen to identify regulators of hepatic lipoprotein secretion. We identified several candidate pathways, including those mediated by Rab11 and Rab8. Surprisingly, inhibition of Rab1b, the major regulator of transport from the endoplasmic reticulum to the Golgi, differently affected the secretion of the very-low-density lipoprotein components ApoE and ApoB100, despite their final association on mature secreted lipoprotein particles. Since hepatitis C virus (HCV) incorporates ApoE and ApoB100 into its virus particle, we also investigated infectious HCV secretion and show that its regulation by Rab1b mirrors that of ApoB100. These observations reveal differential regulation of hepatocyte secretion by Rab1b and advance our understanding of lipoprotein assembly and lipoprotein and HCV secretion

An open label pilot trial of low‐dose lithium for young people at ultra‐high risk for psychosis

Aim: Lithium, even at low doses, appears to offer neuroprotection against a wide variety of insults. In this controlled pilot, we examined the safety (i.e., side‐effect profile) of lithium in a sample of young people identified at ultra‐high risk (UHR) for psychosis. The secondary aim was to explore whether lithium provided a signal of clinical efficacy in reducing transition to psychosis compared with treatment as usual (TAU). Methods: Young people attending the PACE clinic at Orygen, Melbourne, were prescribed a fixed dose (450 mg) of lithium (n = 25) or received TAU (n = 78). The primary outcome examined side‐effects, with transition to psychosis, functioning and measures of psychopathology assessed as secondary outcomes. Results: Participants in both groups were functionally compromised (lithium group GAF = 56.6; monitoring group GAF = 56.9). Side‐effect assessment indicated that lithium was well‐tolerated. 64% (n = 16) of participants in the lithium group were lithium‐adherent to week 12. Few cases transitioned to psychosis across the study period; lithium group 4% (n = 1); monitoring group 7.7% (n = 6). There was no difference in time to transition to psychosis between the groups. No group differences were observed in other functioning and symptom domains, although all outcomes improved over time. Conclusions: With a side‐effect profile either comparable to, or better than UHR antipsychotic trials, lithium might be explored for further research with UHR young people. A definitive larger trial is needed to determine the efficacy of lithium in this cohort

Structural basis for Zika envelope domain III recognition by a germline version of a recurrent neutralizing antibody

Recent epidemics demonstrate the global threat of Zika virus (ZIKV), a flavivirus transmitted by mosquitoes. Although infection is usually asymptomatic or mild, newborns of infected mothers can display severe symptoms, including neurodevelopmental abnormalities and microcephaly. Given the large-scale spread, symptom severity, and lack of treatment or prophylaxis, a safe and effective ZIKV vaccine is urgently needed. However, vaccine design is complicated by concern that elicited antibodies (Abs) may cross-react with other flaviviruses that share a similar envelope protein, such as dengue virus, West Nile virus, and yellow fever virus. This cross-reactivity may worsen symptoms of a subsequent infection through Ab-dependent enhancement. To better understand the neutralizing Ab response and risk of Ab-dependent enhancement, further information on germline Ab binding to ZIKV and the maturation process that gives rise to potently neutralizing Abs is needed. Here we use binding and structural studies to compare mature and inferred-germline Ab binding to envelope protein domain III of ZIKV and other flaviviruses. We show that affinity maturation of the light-chain variable domain is important for strong binding of the recurrent VH3-23/VK1-5 neutralizing Abs to ZIKV envelope protein domain III, and identify interacting residues that contribute to weak, cross-reactive binding to West Nile virus. These findings provide insight into the affinity maturation process and potential cross-reactivity of VH3-23/VK1-5 neutralizing Abs, informing precautions for protein-based vaccines designed to elicit germline versions of neutralizing Abs