Effective decision-making requires organisms to predict reward values and bias behavior toward the best available option. The mesolimbic dopamine system, including the nucleus accumbens (NAc) shell and core, is involved in this process. Although studies support a role of the shell and core in specific aspects of decision-making , no studies have directly compared dopamine release dynamics in these subregions to cues exclusively signaling the availability of different reward magnitudes. Here, fast-scan cyclic voltammetry was used to compare rapid dopamine release dynamics in the NAc subregions during a magnitude-based decision-making task. Rats learned that distinct cues signaled the availability of either a small or large reward (one or two sugar pellets), and then were given an opportunity to choose their preferred option. We found that peak dopamine release tracked the more preferred (higher-magnitude) option in both core and shell subregions. Critically, however, overall  dopamine release was significantly higher and longer lasting in the shell and tracked the preferred magnitude during the entire cue period. Further, in the shell (not core), dopamine signaling significantly declined immediately at the lever press for reward but increased during the period of reward consumption. Collectively, the results indicate that although dopamine release in both the core and shell are activated by cues signaling the opportunity to respond for rewards of different magnitudes, dopamine release in the shell plays a differential and unique role in tracking information related to the outcome value of reward. The nucleus accumbens (NAc) has been shown to facilitate optimal decision-making; however, the precise role of its primary subregions (shell and core) in outcome-based decision-making is unclear. Here, using voltammetric recording methods in behaving rats, we show preferential real-time dopamine signaling within the NAc shell, compared with the core, during a well-learned magnitude-based decision-making task. Collectively, these findings demonstrate that dopamine in the NAc shell plays a unique role in tracking information related to reward outcome value.Effective decision-making depends on an organism’s ability to predict the outcome of its choices and bias behavior toward the option of greatest value. Value-based decision-making recruits the mesolimbic system, including the nucleus accumbens (NAc) and its dopaminergic input . ElectroThe NAc contains two primary subregions, the shell and core, that differ in their afferent and efferent connections  and subsAlthough these studies link the shell and core to specific aspects of decision-making, no reports have directly compared dopamine release dynamics in these subregions to cues signaling the availability of different reward magnitudes. Here, we used fast-scan cyclic voltammetry (FSCV) to examine how rapid dopamine signaling in the NAc core versus shell encodes information about cues during a magnitude-based decision-making task. Rats learned that distinct cues signaled the availability of either a small or large reward (one or two sugar pellets), and then were given an opportunity to choose their preferred option. We found that peak dopamine release tracked the more preferred (higher-magnitude) option in both core and shell subregions. Critically, however, global dopamine release dynamics were significantly higher and longer lasting in the shell and tracked the preferred reward magnitude during the entire cue period. Further, the shell, but not core, exhibited unique dopamine signaling properties relative to the lever press for reward. These findings indicate that cues signaling the opportunity to respond for rewards of different magnitudes activate dopamine release in both the NAc core and shell; however, dopamine release in the shell plays a differential and unique role in tracking information related to reward value.n = 12) rats were ∼90–120 d old and weighed 275–330 g at the start of experiments. Animals were maintained at no less than 85% of pre-experimental body weights by food restriction, except during the postoperative recovery period, when food was given ad libitum (Harlan Lab Chow). Water was available ad libitum throughout the duration of the experiment. Animal procedures were conducted in accordance with the National Institutes of Health Guidelines for the Care and Use of Laboratory Animals, and were approved by the University of North Carolina, Chapel Hill Institutional Animal Care and Use Committee.Singly housed male Sprague-Dawley  described elsewhere . BrieflyAll behavioral experiments were conducted at least 1 wk postsurgery, and rats underwent similar pretraining before beginning each behavioral task. Here, rats were trained to press two distinct levers in which each response was reinforced on a continuous schedule of reinforcement. Reinforced responses resulted in the delivery of a sucrose pellet to a centrally located food cup. Animals were trained to a criterion of 50 presses on each response lever before moving to the behavioral tasks outlined below.Next, rats were trained on a task that involved three types of contingencies  intermixed within 90 total trials per session. At this stage, only a single sucrose pellet was available for each lever press throughout the session. The first two trial types were classified as forced-choice trials. For one trial type, a single cue light was illuminated for 5 s over one lever, followed by extension of both levers. Responses on the cue light illuminated lever (within 15 s) were immediately reinforced with one sucrose pellet. During the other forced-choice trial type, the cue light over the other lever was illuminated for 5 s, followed by extension of both levers. Responses within 15 s on the cue-associated lever were reinforced as above. For both forced-choice trials, responses on the unsignaled lever were counted as errors and resulted in termination of the house light for the remainder of the trial period, with no reward delivery. During the third trial type, termed free choice trials, both cue lights were illuminated for 5 s, after which both levers were extended, and responses on either lever within 15 s were reinforced with one sucrose pellet. After a press on either lever, both levers were retracted and a sucrose pellet was immediately delivered into the food receptacle. To move on to the next phase of training, rats needed to maintain at least 3 d of stable accuracy (80% correct responses).After reaching accuracy criteria, the reward contingency on one of the levers was altered to reflect the reward magnitude decision-making task. A schematic diagram of the final task design is shown in ad libitum.Rats were deeply anesthetized with a mixture of ketamine hydrochloride (100 mg/kg) and xylazine hydrochloride (10 mg/kg), placed in a stereotaxic frame, and surgically prepared for voltammetric recording as described elsewhere . A guiden = 3), after recording a full session of 90 trials, the electrode was lowered another ∼300 µm until another release site was found, at which point another recording was taken for a session of 90 trials.One week after surgery, rats were food-restricted and retrained on the magnitude task until they reached presurgery performance (maximum of five sessions). Changes in dopamine concentration during the task were assessed using FSCV as described elsewhere . On the After completion of the experiments, rats were deeply anesthetized with a mixture of ketamine (100 mg/kg) and xylazine (10 mg/kg). To mark the placement of the electrode tip, a tungsten electrode housed in a micromanipulator was lowered to the recording site, and a small electrolytic lesion was made. Brains were then extracted, sliced on a freezing cryostat, and placed onto slides. The location of electrode tips was assessed by visual examination of successive coronal sections in comparison to visual landmarks and the anatomic organization of the NAc core and shell, as represented in a stereotaxic atlas.t tests were used to compare accuracy  and percentage errors during high and low reward trial types. For free choice trials, a paired t test was used to compare reward magnitude preferences.All statistical analyses are reported in Analysis of FSCV recordings was similar to that of previous reports . Brieflyt tests. Likewise, we used a similar approach to examine cue-evoked dopamine release during forced choice correct versus incorrect  trials. Here, we included equal numbers of correct and incorrect trials  and compared peak dopamine release using paired t tests.Because free choice trials allowed rats to choose a large-versus-small sucrose reward, there were unequal numbers of responses to high- versus low-magnitude options. Thus, because of differences in variance, it would be inappropriate to compare dopamine release for the average of 26 high-magnitude choices compared with 4 low-magnitude choices. Therefore, we combined all dopamine concentrations for the 30 free choice trials, regardless of what option the rat eventually chose. To confirm this approach, we randomly selected an equal number of free choice trials in which animals responded to the low versus high choice  and compared peak dopamine release during these trials using paired To assess the differential effects of the three cue types on dopamine release in both core and shell, peak dopamine concentrations within 2 s following cue presentation were analyzed using a two-way ANOVA with Bonferroni’s multiple comparison test. To compare amounts of dopamine release in the shell versus core, we examined the cumulative dopamine release in each region by summing the concentration of dopamine in each bin during 0.5–4 s after cue onset for the forced and free choice trials to provide an estimate area under the curve and completed a two-way ANOVA with Bonferroni’s multiple comparison test on these data. To examine dopamine release dynamics relative to the lever press, we examined the depth of the dopamine “trough” after the press. This was analyzed by taking the minimum dopamine concentration within 2 s after lever press in the core and shell and performing a two-way ANOVA. To analyze dopamine release after lever press, a one-way ANOVA with Fisher’s LSD multiple comparison test was performed comparing dopamine concentration at the peak trough time point compared with 2 s afterward at the peak of the increase.All analyses were considered significant at α = 0.05. Statistical and graphical analysis were performed using GraphPad Prism 6.0 for Windows (GraphPad Software) and Neuroexplorer for Windows version 4.034 (Plexon Inc.). Statistical outliers were calculated using extreme studentized deviate test and excluded from further analysis.t14 = 3.726, p < 0.01a) and made significantly more errors  during low forced choice trials. On free choice trials, all rats exhibited a significant preference (>60% choice) for the high-magnitude option over the low-magnitude option .Rats rapidly learned the magnitude decision-making task and discriminated between the cue types. Specifically, rats completed significantly more correct responses during high forced choice trials .Reward-predictive cues evoked rapid dopamine release in the NAc shell and core. However, forced choice high-magnitude trials induced higher concentrations of dopamine release than forced choice low-magnitude trials. This finding is illustrated for a representative animal in t5 = 1.779, p > 0.05d) or core . Free choice trials were combined regardless of rats’ eventual choice, as random sampling analysis indicated no difference in dopamine release during trials in which rats chose the high versus low option in either the shell  or core . It is immediately apparent that although dopamine was released to cues in both NAc subregions, it was generally higher in concentration in the shell  and a main effect of region , but importantly, no significant interaction between region and trial type . These findings confirm that although all cue types significantly increased dopamine release in both regions, release was significantly higher in the NAc shell, compared with core. Further, dopamine signaling during high forced choice and free choice cues was significantly higher than during low forced cues across both subregions (p < 0.05f).he shell A, left the shell A, right.he shell A. Two-waF2,26 = 7.584, p < 0.01g) but no significant main effect of region  or interaction . Post hoc tests on the main effect of trial type revealed higher peak dopamine release for high and choice trials, compared with low magnitude trials, independent of region (p < 0.05g). High and choice trial peak dopamine were not different from each other (p > 0.05g).Reward-predictive cues also evoked significant increases in peak dopamine release in both subregions, as illustrated in F1,12 = 5.744, p < 0.05h) but no main effect of trial type  or interaction . These findings indicate that the dip in dopamine evident after the lever press was most pronounced in the shell, regardless of trial type. Interestingly, in the shell, a subsequent increase in peak dopamine release was observed within 2 s after the lever press , but only in forced high and free choice trials (p < 0.05d). This increase was not observed in the core .Five seconds after cue presentation, both levers were extended into the chamber and rats could press a single lever indicating either one reward outcome or no reward  or choice of high versus low reward options . In the shell, a pronounced dip in dopamine release occurred after lever press A, left. Placement of carbon fiber electrode tips used in FSCV experiments were confirmed to be in the NAc core or shell .In the present study, electrochemical methods were used in rats to compare dopamine signaling dynamics in the NAc shell and core during a magnitude-based decision-making task in which discrete cues signaled the availability of different reward sizes (one vs. two sugar pellets). Although similar cue-associated peak dopamine release events were observed across the two NAc subregions, higher dopamine concentrations that extended throughout cue periods, and tracked predicted magnitude information was exclusively measured in the shell. Further, in the shell (not core), there was a significant decline in dopamine signaling immediately after the lever press for reward followed by a significant increase in dopamine release at the time of reward consumption. Collectively, the present findings provide novel insight into rapid dopamine signaling dynamics across NAc subregions and reveal a unique role of the shell in tracking information about expected outcome value.Elements of value-based decision-making may be separable into two interrelated but distinct components , includiSome previous studies have reported that cue-associated rapid dopamine release in the NAc core encodes differences in reward magnitude . HoweverUsing this comparative approach, we were able to reveal that although peak dopamine release to cues was similar across the shell and core, overall dopamine release was significantly larger and longer lasting in the shell , compared with the core. Further, a brief yet significant dip in dopamine release where concentrations extended down to baseline  was evident immediately after the lever press in the shell during all trial types . This significant decline in dopamine release was not observed in the core. Although it is not known exactly what this postresponse drop in dopamine release reflects, it may be indicative of a behavioral shift from food seeking to consummatory behavior. Indeed, electrophysiological studies have shown that neurons in the NAc shell, but not core, were inhibited on lever press for a reward , and thaThe present findings are also consistent with previous studies implicating the shell in encoding reward outcome . For exaOur findings are also consistent with previous work by In conclusion, the present study shows that peak cue-evoked dopamine release in both core and shell subregions scaled to information related to rewards of different magnitudes. However, throughout the entire 5-s cue duration, shell dopamine continued to track magnitude information and remained increased throughout the entire cue period, reflecting the expected value of the predicted reward. Further, a pause in dopamine signaling was observed in the shell (not core) immediately at the lever press for reward, followed by a brief increase in dopamine release during reward delivery and consumption. This unique signal pattern likely relates to the shell’s unique role in consummatory behaviors. Collectively, the results support the view that the core encodes individual, preferential value and is better linked to tracking simple outcome value. Future studies  are needed to determine whether the unique dopamine signaling properties in the NAc shell observed in this study are causally linked to behaviors involving simple decisions to respond for rewards of different magnitudes.


The aim of this study was to investigate whether the background intakes of total dietary fat, monounsaturated fatty acids (MUFA) and polyunsaturated fatty acids (PUFA) modulate the effects of dietary saturated fatty acids (SFA) on metabolic syndrome (MetS).This population-based cross-sectional study was conducted on a representative sample of 4 677 adults, aged 19 to 84 years. MetS was defined according to the ATP III criteria.Median intakes of SFA, MUFA and PUFA were 9.5, 9.6 and 5.6% of total energy. High SFA intakes were associated with higher prevalence of MetS, in both individuals with higher and lower median intakes of total fat, MUFA and PUFA.Our findings indicate that SFA intakes were positively associated with the prevalence of MetS, independent of total dietary fat, MUFA and PUFA intake. Metabolic syndrome (MetS), a constellation of metabolic abnormalities including glucose intolerance, abdominal obesity, dyslipidemia and hypertension , is highHowever, previous studies have shown no significant interaction between dietary SFA and PUFA on cardiometabolic risk factors , 11. In n = 33), stroke (n = 6), cancer (n = 7), reported daily energy intakes outside the range of 800–4200 kcal/d (n = 322) and for missing data on physical activity, or any of anthropometrical measurements, and biochemical variables (n = 92). Also excluded were participants with hyperlipidemia, hyperglycemia and hypertension that had changed their dietary intake (n = 182). Finally, data for 4677 individuals  were analyzed . Serum triglyceride concentrations were measured using triglyceride kits  by the enzymatic calorimetric test with glycerol phosphate oxidase. HDL cholesterol was measured after precipitation of the apolipoprotein B-containing lipoproteins with phosphotungistic acid. Serum fasting glucose concentration was assayed using an enzymatic colorimetric method with the glucose oxidase technique. Inter- and intra-assay coefficients of variations were both 2.2 % for serum glucose, 2 and 0.5 % for HDL-C and 1.6 and 0.6 % for triglycerides, respectively.2). Waist circumference (WC) was measured at the level of the umbilicus site, using an outstretched tape meter, without pressure to body surfaces and was recorded to the nearest 0.5 cm.Weight was measured while the subjects were minimally clothed and not wearing shoes, using digital scale and was recorded to the nearest 100 g. Height was measured while subjects were standing without shoes, with their shoulders in a normal position, using a tape fixed to the wall and was recorded to the nearest 0.5 cm. Body mass index (BMI) was calculated as weight (kg) divided by square of height  was used for all statistical analyses. Significant differences in characteristics and dietary intakes in the categories of total fat intake and SFA using joint categories of higher (≥ median) versus lower (< median) intakes were evaluated using one-way analysis of variance for continuous variables and values were reported as Mean (SD). Chi-square test was used to detect any significant differences in the distribution of participants across quartile categories with regard to qualitative variables.Odds Ratio (ORs) and their 95 % confidence intervals was estimated for the MetS and its components, according to both unsaturated fatty acids (MUFA and PUFA) and SFA using joint categories of higher median intake versus lower median intakes , by mult2. The reported mean (SD) daily intakes were: Total fat 29.9 (6.1) % of total energy intake; SFA 10.2 (2.8) % of total energy intake; MUFA 10.2 (2.8) % of total energy intake and PUFA 6.1 (1.9) % of total energy intake. Baseline characteristics and dietary intakes of the participants according to both total fat and SFA intakes are shown in Table Of 4677 study participants, 44.4 % were male and 55.6 % were female, with a mean age (SD) of 41.7 (13.9) years and had mean BMI of 27.2 (4.7) kg/m22 % higher among subjects with a higher total fat intake (95 % CI: 1.05-1.55), compared with subjects with lower intakes of both. Also the combination of a high SFA and total fat intakes was associated with abnormal glucose homeostasis, elevated blood pressure, and high serum triglyceride concentrations after adjustment for confounding factors.Table ORs of the MetS and its components according to intakes of both MUFA and SFA is shown in Table Table The aim of the current study was to investigate whether the background intake of total dietary fat, MUFA and PUFA modulates the effects of SFA on the MetS and its components. Our findings showed that among Tehranian adults, dietary SFA was associated with high prevalence of the MetS and intake of total dietary fat, MUFA and PUFA (higher or lower than the median) did not affect the this association. Also the combination of a high SFA and total fat intakes was associated with abnormal glucose homeostasis, elevated blood pressure, and high serum triglyceride concentrations after adjustment for confounding factors.In prospective studies, dietary SFA has been considered a cardiometabolic risk factors , 6, but Results on the replacement of MUFA with SFA on cardiometabolic risk factors were inconsistent , 24–26. Our findings show high prevalence of MetS and abnormal glucose homeostasis among subjects with high intake of SFA, independent of dietary PUFA. Consistent with our results, epidemiologic and experimental studies have shown no significant interaction between SFA and PUFA , 11. In Our study has several strengths. In the current study, we evaluate whether total dietary fat, PUFA and MUFA intakes influence relations between SFA and Mets and its components. We found that the association between the SFA intake and some components of MetS is modulated by total dietary fat, as suggested by some previous studies , 27. AlsSome limitations should be considered, one being the use of UASD FCT to determine the intakes of PUFA, MUFA and SFA, because of not having a complete Iranian FCT. Given the cross-sectional design, we could not determine causality between interaction of different type of fatty acids and the MetS and its components. Future studies using longitudinal data are needed to determine these effects. In addition, this study included only healthy adults, and our findings cannot be extrapolated to other populations.Our findings indicate that SFA intakes were positively associated with the prevalence of MetS, independent of total dietary fat, MUFA and PUFA intake. However total dietary fat may modulate the association between SFA and some components of MetS, results emphasizing that both the quality and quantity of dietary fat are relevant with Mets and cardiometabolic risk factors.


Progesterone is a potent drug for synchronization of the estrus and ovulation cycles in bovine. At present, the estrus cycle of bovine is controlled by the insertion of progesterone-embedded silicone bands. The disadvantage of nondegradable polymer inserts is to require for disposal of these bands after their use. The study currently focuses on preparation of biodegradable progesterone-incorporated nanofiber for estrus synchronization. Three different concentrations  of progesterone-impregnated nanofibers were fabricated using electrospinning. The spun membrane were characterized by scanning electron microscopy, X-ray diffraction, differential scanning calorimetry, thermogravimetric analysis, and Fourier transform infrared spectroscopy. Uniform surface morphology, narrow size distribution, and interaction between progesterone and zein were confirmed by SEM. FTIR spectroscopy indicated miscibility and interaction between zein and progesterone. X-ray analysis indicated that the size of zein crystallites increased with progesterone content in nanofibers. Significant differences in thermal behavior of progesterone-impregnated nanofiber were observed by DSC. Cell viability studies of progesterone-loaded nanofiber were examined using MTT assay. In vitro release experiment is to identify the suitable progesterone concentration for estrus synchronization. This study confirms that progesterone-impregnated nanofibers are an ideal vehicle for progesterone delivery for estrus synchronization of bovines. Electrospinning is a technique used to form nanoscale fibers. It is quite versatile for fabricating nanofibers from various synthetic or natural polymers . In liteZein is a mixture of proteins with different molecular weights in corn gluten. Apart from biodegradability and biocompatibility, zein has low hydrophilicity, high elasticity, and film-forming capabilities, and it is considered a potential raw material for bioengineering application .The artificial induction and synchronization of estrus in production animals is critical to ensure a positive balance of the cost-benefit equation of the artificial insemination related activities. The usual administration of hormones must be very precise. The controlled hormone release is a current technological challenge. One interesting agent to be tested in such delivery system is the progesterone, a steroid hormone naturally produced by the corpus luteum of the ovaries of mammals and involved in their pregnancy. In veterinary medicine, exogenous progesterone is used as a potent drug for suppression of estrus and ovulation, making possible the synchronization of the estrus and ovulation cycles in livestock animals . In thisZein from corn and progesterone were purchased from Sigma-Aldrich (USA). Ethanol 99.7% purity was supplied by Merck−1. Electrospinning processes were carried out under ambient conditions (24 ± 3 °C with relative humidity 57 ± 4%) [Zein was dissolved in ethanol and kept under vigorous stirring overnight at room temperature. Various concentrations  of progesterone were dissolved in ethanol for an hour at room temperature. Both solutions were mixed for an hour. Progesterone-loaded zein fibers prepared by electrospinning were spun using a voltage of 24 kV, working distance of 12 cm, and feed rate of 2 μL min57 ± 4%) .Scanning electron microscopy is used to check the surface morphology of three different concentrations  of progesterone-incorporated nanofiber. The SEM characterization of electrospun nanofiber was performed using JEOL JSM-6480 V (accelerative voltage 20 kV) scanning electron microscopy at the Nanotechnology Department of SRM University, Chennai. The nanofiber samples collected on the aluminum foil was peeled out and then mounted on SEM sample holder using graphite-impregnated adhesive conductive black carbon tape, coated with platinum, and visualized under SEM at various magnifications.−1 with incident beam.XRD patterns were generated from nonwoven fibrous mat using a Rigaku D/Max ULTIMA 11 X-ray diffractometer (Japan). The X-rays are generated by a cathode ray tube filtered to produce monochromatic radiation directed towards the sample. The interaction of the incident rays with the sample produces constructive interference (and diffracted rays). The diffracted intensity were recorded from 0 to 1400 at 2θ angle and the pattern was recorded by Cu K radiation with 1.5418 Å and graphite monochromatic filtering wave at a tube voltage of 40 kV and tube current of 30 mA, and scanning in the region of 0 to 70 at 6 min2 atmosphere 80 L/min). The glass transition temperature (Tg) was evaluated with the Stare-software version 6.01 . Zein films were stored over silica gel or at different relative humidities for 5 days prior to measurement to achieve different water contents. The relative humidity (r.h.) was controlled by saturated salt solutions  The predicted Tg values were calculated with the Gordon-Taylor-equation.Differential scanning calorimetry (DSC) measurements  were performed on samples of 5 mg in the range of −100 to 200 °C at a heating rate of 10 °C/min  were recorded in the range 4000–400 cm3 cells/well were seeded in 96-well plates containing DMEM and incubated in 5% CO2 at 37 °C for 24 h. The medium was changed after 1 h and 100 μL of different concentrations  of the 1.2 g progesterone-loaded nanofiber dissolved with PBS was added to the wells and incubated for 24 h at 37 °C in the CO2 incubator. One hundred microliter of MTT (5 mg/mL) was added to the wells containing cells and nanofibers of different concentrations. It was incubated at 37 °C for 4 h. The medium was then removed and 20 μL of DMSO was added to the wells. It was then shaken and incubated at 37 °C for 15 min and the absorbance was measured at 575 nm.Vero cells from ATCC are used for the MTT assay. One hundred-microliter Vero cells at the concentration of 3 × 10The in vitro release studies were performed at three different progesterone concentrations  of nanofibers in a shaker at 37 °C. A weighed quantity of the fibers (20 mg) was suspended in PBS of pH 7.4. Then, it was kept in a shaker for seven days at 37 °C. The sample was withdrawn at regular one day intervals up to 7 days and replaced with the same volume of freshly prepared PBS pH 7.4. The withdrawn samples were used for OD measurement at 237 nm by a UV-visible spectrophotometer .In this, the electrospinning of zein nanofibers was mostly carried out by using ethanol system which resulted in ribbon-like fiber morphology due to the rapid mat formation of the fiber core because of the very fast evaporation of the solvent , 16.The fibers were spun from the same polymer solution and under the same spinning conditions with different concentrations of progesterone and were characterized by SEM. Zein nanofiber without progesterone had ribbon morphology with smooth surfaces and uniform structures Fig.  comparedThe XRD patterns of electrospun zein nanofibers and different concentration of progesterone loaded nanofibers are shown in Fig. DSC was done on the electrospun nanofiber and progesterone incorporated nanofibers in order to determine the thermal behavior of the nanofibers. The glass transition temperature (Tg) of the different samples are presented in Fig. −1 (Fig. −1 (Fig. FTIR spectra of electrospun zein nanofibers and progesterone loaded nanofibers were shown in Fig. −1 Fig. . These b −1 Fig. . These bMTT assay was the most efficient, due to a less experimental error. The graph in Fig. The percentage release of progesterone was also estimated for different concentrations of progesterone loaded and expressed in Fig. The results of the current study confirm some miscibility of progesterone on hydrophobic biopolymers according to SEM, XRD, DSC, TGA, and FTIR. The electrospinning can be appropriately used to encapsulate active agents in biodegradable and biocompatible polymers, providing a hormone release sustainably. The increases in the concentration of progesterone affect the nanofiber size and morphology was confirmed by SEM. Progesterone at various concentrations did not affect the structural integrity of the nanofibers. Progesterone was found to have the effect of plasticizer when added to zein polymer. The electrospinning can be appropriated used to encapsulate active agents in biodegradable and biocompatible polymers, providing a hormone release sustainably. This study clearly indicated that 1.2 g progesterone-loaded zein nanofibers can be potentially used in controlled delivering of progesterone, in livestock animals for estrus synchronization.


Vibrio vulnificus is a causative agent of fatal septicemia and necrotic wound infection and the pathogen infection became an important public health problem in many counties. Vibrio vulnificus causes RtxA1 toxin-induced acute cell death. We tried to identify natural products that inhibit the acute cytotoxicity of V. vulnificus using a lactate hydrogenase assay. A polyphenol pyrogallol protected HeLa cells from V. vulnificus-induced cytotoxicity. Pyrogallol also decreased the growth of V. vulnificus; this inhibitory effect was more significant during log phase than stationary phase. To further elucidate the inhibitory mechanism, pyrogallol-induced toxicity was compared between a V. vulnificus catalase-peroxidase mutant (katG−) and the isogenic wild-type MO6-24/O strains. No growth was observed for the katG− mutant in the presence of pyrogallol (50 μg/mL) even after 24 h, whereas the wild-type strain demonstrated growth recovery following a prolonged lag phase. Pyrogallol-mediated growth inhibition of the katG− mutant strain was partially rescued by exogenous catalase treatment. These results indicate that the mechanism by which pyrogallol inhibits the growth and cytotoxicity of V. vulnificus likely involves polyphenol-induced prooxidant damage. Taken together, these results suggest that pyrogallol has potential for development as a new paradigm drug to treat infectious diseases. Vibrio vulnificus is a halophilic, estuarine bacterium that causes fatal septicemia and necrotic wound infections [V. vulnificus infection exhibits a broad pathogenic spectrum, a fulminating course, and a high mortality rate (> 50%) with death occurring within days [Vibrio infections [V. vulnificus is repeats-in-toxin A1 (RtxA1), which induces the programmed necrotic death of host cells [V. vulnificus infection, we conducted a screen of various natural products [V. vulnificus pathogenesis by interfering with its adhesion to host cells and inhibiting the production of RtxA1 [Vibrio species [Helicobacter pylori urease and Vibrio harveyi quorum sensing [V. vulnificus. In addition, we evaluated pyrogallol-induced toxicity in wild-type (wt) V. vulnificus and in a catalase-peroxidase (katG−) mutant strain.fections , 2. V. vhin days . Thus, tfections . Previouy rate > 0% with dst cells . To idenproducts , 8 and fof RtxA1 . Polyphe species . The hea species . A polyp species . The pro species . Accordi sensing . In the V. vulnificus strains were grown in heart infusion (HI) broth  in a shaking incubator at 200 rpm and 37°C. V. vulnificus MO6-24/O, a clinical isolate was provided by J. Glenn Morris, Jr., of the University of Maryland [V. vulnificus pathogenesis study and the complete genome sequence was annotated [katG−) was constructed in V. vulnificus wild type (wt) strain MO6-24/O using a counter-selection strategy and the suicide vector pKAS32 [katG gene composed of 2,172 nucleotides. Pyrogallol, catalase, and hydrogen peroxide (H2O2)  were dissolved in phosphate-buffered saline (PBS).Maryland . MO6-24/nnotated . A catalr pKAS32 , 16. Pol2 atmosphere.HeLa cells  were cultured in Dulbecco's Modified Eagle's Medium  supplemented with 10% heat-inactivated fetal bovine serum  at 37°C in an incubator with a 5% CO4 cells/well) and cultured overnight. The cells were treated with serum-free DMEM plus pyrogallol (50 μg/mL) for 1 h prior to V. vulnificus infection at a multiplicity of infection (MOI) of 20 for 2 h. The HeLa cells were stained with a Trypan blue solution (Sigma-Aldrich) for 10 min, followed by washing with PBS. The Cell images were acquired using an inverted microscope with a digital camera .HeLa cells were seeded in a 48-well cell culture plate  as previously described [4 cells/well) and cultured overnight. The cells were then washed with serum-free DMEM and treated with pyrogallol or H2O2 for 1 h prior to V. vulnificus infection at an MOI of 20 for 2h. Lactate dehydrogenase (LDH) released into the supernatant was assayed as a cytotoxicity marker in accordance with the manufacturer’s protocol.escribed , 5. HeLa2 for 24 h. Cell viability was measured using a 3--5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) assay  in accordance with the manufacturer’s protocol.HeLa cells cultured in 96-well microplates overnight were washed with serum-free DMEM and treated with pyrogallol (20–100 μg/mL) in a 37°C incubator with 5% CO2O2 (0.25–1.0 mM) in a 37°C incubator with 5% CO2 for 2 h and cytotoxicity was measured using CytoTox96 as described above.HeLa cells were treated with HV. vulnificus wild type or the katG− mutant strains were grown in a shaking incubator at 37°C overnight and the culture suspensions were diluted 1:1000 into fresh HI broth. The diluted bacterial suspensions were inoculated into 96-well microplates with pyrogallol (20–100 μg/mL) or catalase (10 μg/mL) and then incubated at 37°C for 6 h. Bacterial growth was determined by measuring the absorbance at 600 nm using a SpectraMax 190 microplate reader . To assay bacterial growth in a shaking incubator for 24 h, the diluted suspensions of V. vulnificus were cultured in 50-mL tubes and the optical density was measured every 3 h using a Biophotometer .2O2 (0.25–1.0 mM) on V. vulnificus viability, approximately 1 × 107 colony forming units/mL (CFU/mL) of V. vulnificus wt or the katG− mutant strains were cultured in DMEM for 2 h. For enumeration of live bacterial cells, the V. vulnificus culture suspensions were 10-fold serially diluted with PBS. The serial dilutions (10 μL) were loaded on HI agar plates and incubated in a 37°C incubator overnight.To test the effect of HV. vulnificus katG gene was amplified by PCR using a DNA polymerase  and cloned into the pGEX-4T expression vector . The constructed plasmid was transformed into E. coli DH5α and the glutathione S-transferase (GST)-fusion protein was induced according to the manufacturer’s protocol (Amersham Pharmacia Biotech). The resulting GST-KatG fusion protein was purified by affinity chromatography in accordance with the manufacturer's recommendations (Amersham Pharmacia Biotech). A polyclonal antibody against the GST-KatG fusion protein was produced using New Zealand white rabbits according to previously described methods [A DNA fragment encoding the  methods .V. vulnificus wt and the katG− mutant strains were cultured in HI broth with or without pyrogallol (50 μg/mL) or catalase (10 μg/mL) in a 37°C shaking incubator for 6 h or 24 h. For KatG protein analysis, the bacterial pellets (2 × 108 CFU) were suspended in sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) sample buffer and boiled in a water bath for 10 min. For RtxA1, 300 μL supernatants from bacteria cultured in HI for 6 h were concentrated using cold acetone. RtxA1 was detected using an anti-rabbit RtxA1 antibody specific to RtxA1-C [ RtxA1-C . QuantitStatistical differences were evaluated using one way ANOVA. All experiments were repeated three times and the results shown are from representative experiments. All results are presented as the means ± standard error of the mean (SEM).V. vulnificus is highly toxic to host cells; this cytotoxicity is caused mainly by the RtxA1 toxin [V. vulnificus cytotoxicity, several natural products were tested using the Trypan blue staining method. Treatment with pyrogallol (50 μg/mL) prevented HeLa cells from becoming damaged by the pathogen  significantly inhibited V. vulnificus growth in a dose-dependent manner  on V. vulnificus growth was more significant during log phase (3–6 h) than in stationary phase  after 24 h, whereas the wild type strain showed growth recovery in stationary phase  did not show any growth in the presence of pyrogallol (V. vulnificus cells [V. vulnificus during stationary phase (V. vulnificus wt and the katG− mutant strain (2O2 has been suggested as an important beneficial mechanism. Our results show that H2O2 exhibits inhibitory effects on V. vulnificus growth (V. vulnificus growth via the production of reactive oxygen species. The growth inhibition in HI broth culture by pyrogallol was more significant in log phase than in stationary phase (Pyrogallol has been reported to possess both antioxidant and prooxidant properties . In supprogallol . The Katus cells . Here, wry phase  and in ary phase . Additiot strain . In parts growth  similar s growth . These rry phase . Pyrogalry phase , which m


Dopamine-modified alginate and gelatin were prepared. The polymers were characterized and the properties of their aqueous solutions were investigated. Aqueous solutions of dopamine-modified alginate and gelatin with a concentration exceeding 20 mg/mL naturally formed gels after 16 h. Although polydopamine itself was not used for template-assisted nanostructure formation, the modified polymers could be used with dopamine. Mixing with dopamine allowed the precise shape of the template to be maintained in the resulting material, allowing nanopatterned surfaces and nanotubes to be prepared. In 2007, it was reported that dopamine can be used to modify various surfaces including both inorganic and organic ones  using a Some modifications have been devised to circumvent the mechanical fragility of PDA films. The first approach is layer-by-layer formation ,27, and In this study, dopamine-modified alginate and gelatin were prepared. The polymers were characterized by oxidation, gelation, and film formation. Finally, template-assisted nanostructures were prepared using these modified polymers. N-hydroxysuccinimide (NHS) and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC) were purchased from Tokyo Chemical Industry Co., Ltd. .Chemicals were used as received without further purification. Dopamine hydrochloride and gelatin (from porcine skin Type A) were purchased from Sigma-Aldrich . Sodium alginate (80–120 mPa·s) was purchased from Wako Pure Chemical Industries, Ltd. . w/v). EDC and NHS were then added to the alginate solution at a molar ratio equal to that of alginate and reacted at room temperature for 15 min. Dopamine hydrochloride was added to the solution at a 3:1 molar ratio of catechol groups to carboxyl groups. The reaction was performed at room temperature for 12 h. The reaction mixture was dialyzed using DPBS (pH = 4) and acidic distilled water (pH = 5) for 24 h with a dialysis membrane , and subsequently freeze-dried. Gelatin was dissolved in DPBS (pH = 4) at 40 °C, and then a gelatin-catechol (GC) conjugate was synthesized using the method described above. The preparation process of the dopamine-modified polymers is shown in 1H nuclear magnetic resonance (NMR) measurements on a 400 MHz NMR spectrometer . The substitution degrees of catechol groups for carboxyl groups in AC and GC were determined by measuring the absorbance at 280 nm using a Ultraviolet-visible (UV–vis) spectrophotometer   because the aromatic ring structure of conjugated catechol group exhibits an absorption peak at this wavelength [Syntheses of AC and GC were confirmed by velength . DopaminAC and GC  were dissolved in 10 mM Tris-HCl buffer  to make solutions with various concentrations. The solutions were incubated at room temperature, and photographs were taken after prescribed periods.AC and GC (5 mg/mL), dopamine hydrochloride (1 mg/mL), and a total of 1 mg/mL of GC mixed with dopamine hydrochloride at weight ratios of 10:1 (denoted as GC-dopamine 10:1) and 1:1 (denoted as GC-dopamine 1:1) were dissolved in 10 mM Tris-HCl buffer (pH = 8.5). Polycarbonate membranes  were dipped in the solutions. After incubating at 60 °C for 8 h (7 h for GC-dopamine 10:1 and GC-dopamine 1:1) at a shaking speed 100 rpm, the membranes were rinsed with the same buffer several times to remove the aggregated particles and then washed at 40 °C three times at a shaking speed 80 rpm for 10 min each time. The membranes were then dried under low vacuum conditions.Glass substrates (diameter: 15 mm) were cleaned by sequential sonication in 6 M HCl and then acetone for 10 min each time. Ultrapurified water (20 µL) from Milli-Q purifier  was then dropped on each glass substrate and a piece of polycarbonate membrane (around 2 mm × 3 mm rectangle) coated with AC, GC, dopamine, GC-dopamine 10:1, or GC-dopamine 1:1 was placed on the Milli-Q drop with the film facing the glass. The glass was then dried in air. Dimethylformamide (DMF) (100 µL) was dropped on the membrane and incubated at room temperature for around 5 min to dissolve the polycarbonate membrane before discarding; this dissolving process was repeated six times. Finally, the DMF was removed by rinsing the film with acetone and drying in air. The thickness of the film transferred to the glass substrate was measured with a reflection confocal laser microscope  by scratching the film with tweezers to expose the glass substrate.The nanopattern film fabrication process is illustrated in As shown in 1H NMR analysis as shown in 1H NMR analysis ; equimolar amounts of NaIO4 and catechol in AC resulted in the formation of a hydrogel within 3–4 min after induction of gelation [The changes of aqueous solutions containing the prepared conjugates are illustrated in gelation . The hydFilms were formed using concentrations that did not cause gel formation, and their thicknesses were measured. Substrates were dipped in AC and GC (5 mg/mL), and dopamine (1 mg/mL) was used as a control a. After Substrates were also dipped in GC (1 mg/mL), and a total concentration of 1 mg/mL of GC mixed with dopamine at weight ratios of 10:1 (denoted as GC-dopamine 10:1) and 1:1 (denoted as GC-dopamine 1:1) b. After The morphologies of the patterned films produced under each sets of conditions are shown in Two dopamine-modified polymers were synthesized by EDC/NHS chemistry and their oxidation, gelation, and film formation characteristics were determined. With the assistance of templates, patterned films with a channel structure and nanotube arrays were fabricated by combining dopamine with the dopamine-modified polymers. These combinations provided sufficient stiffness and flexibility to form various templated nanostructures, which was difficult with either dopamine or a dopamine-modified polymer alone. The present material design will enable us to fabricate various nanostructures for specific applications.


The cell surface molecule CD99 has gained interest because of its involvement in regulating cell differentiation and adhesion/migration of immune and tumor cells. However, the molecule plays an intriguing and dual role in different cell types. In particular, it acts as a requirement for cell malignancy or as an oncosuppressor in tumors. In addition, the gene encodes for two different isoforms, which also act in opposition inside the same cell. This review highlights key studies focusing on the dual role of CD99 and its isoforms and discusses major critical issues, challenges, and strategies for overcoming those challenges. The review specifically underscores the properties that make the molecule an attractive therapeutic target and identifies new relationships and areas of study that may be exploited. The elucidation of the spatial and temporal control of the expression of CD99 in normal and tumor cells is required to obtain a full appreciation of this molecule and its signaling. CD99 gene encodes two distinct proteins as a result of the alternative splicing process of the cytoplasmic region: a wild-type full-length CD99, or CD99 type I (CD99wt), containing 185 aminoacids (corresponding to a molecular weight of 32 kDa), and a truncated form, or CD99 type II (CD99sh), containing 161 aminoacids (corresponding to a molecular weight of 28 kDa) [CD99, a glycosylated transmembrane protein, is involved in many essential cellular functions. For example, cell adhesion and migration, cell death and differentiation, intracellular protein trafficking, endocytosis and exocytosis are influenced by the CD99-regulated signaling.  28 kDa) . The CD9 28 kDa) ,21,22. TThe opposite, dual functions of CD99 isoforms.. CD99 hview see ,34,35 anview see ,37, apopview see ,38, adheview see ,34,39,40view see ,42, and view see ,44,45,46view see ,44,45 anview see : thus, CStrong expression of CD99 has also been reported in immature basal keratinocytes , in osteMIC2 gene can positively or negatively regulate cellular adhesion, apoptosis, migration, and metastasis. Indeed, CD99sh frequently plays a negative role in CD99wt-dependent functions, counteracting its effects [The differential expression levels and functions of alternative splicing products are among the physiological and pathological features of many cell adhesion molecules. Regarding CD99, several studies have shown that, by encoding for CD99wt and/or CD99sh, the  effects ,22. It h effects , leading effects . In cont effects . Transfe effects . When co effects . However effects ,52,53. T effects , whereas effects .CD99 isoforms also dictate different functions in malignancy, exerting opposite effects on crucial biological processes, such as migration/invasion, growth in anchorage-independent conditions, and differentiation and metastasis in osteosarcoma, prostate cancer, and breast carcinoma ,22,55. FHowever, with its short cytoplasmic tail, it is unlikely that CD99 itself takes part in signaling events. The functions of CD99 may depend on its association with other membrane proteins, possibly explaining the dual roles of CD99 and its isoforms in various cell types.EWS gene-rearrangement with FLI1, ERG or, in rare cases, other ETS genes [CD99 promoter [High expression of CD99 is a distinctive feature of Ewing sarcoma. Moreover, the detection of CD99 is routinely used for the differential diagnosis of conventional Ewing sarcoma with respect to other types of small round-cell tumors of childhood, including lymphomas, small round-cell osteosarcoma, and the most recently defined Ewing-like round cell tumors, a group of very rare malignancies morphologically resembling Ewing sarcoma but carrying a different type of genetic alterations and exhibiting lower and dispersed expression of CD99 ,61,62,63TS genes ,65 are tTS genes ,67 whileTS genes . The preTS genes ,69. Thespromoter ,59 or inpromoter . From a promoter ,71, and promoter ,56,59. Ppromoter ). This fpromoter . Therefopromoter . Notablypromoter , confirmSimilar evidence has been obtained in hematopoietic tumors. CD99 is highly expressed in T-lineage acute lymphoblastic leukemia (T-ALL) , early BThe inverse relation between CD99 expression and differentiation has also been confirmed in thymic tumors, in which CD99 is useful for identifying immature T cells and for distinguishing thymoma from thymic and non-thymic carcinomas .CD99 is also upregulated in tumors of the nervous system. CD99 was found to be highly expressed in ependymomas ,79, in mTaken together, these findings suggest that CD99 may play an important role in the migration and invasion of certain human tumors and significantly affect crucial signaling pathways, such as AKT, ERK, and Rho/Rac signaling pathways. However, even if the final effect of CD99 overexpression is the same in all these tumors, the intracellular mechanisms of action are far from being commonly defined and strictly cellular context-dependent.In Hodgkin’s lymphomas , osteosaMethylation of the CD99 promoter, CD99 loss of heterozygosity (LOH), and loss of SP1 expression, which is a potent positive regulator of CD99, can account for CD99 downregulation in most cases ,85. HoweFrom a mechanistic point of view, CD99 activity with respect to cell migration and invasion is at least partly related to the regulation of the cytoskeleton, actin remodeling, and increased expression of certain metalloproteinases, such as MMP-2 . In osteIn other tumors, however, the effect of CD99 on cytoskeletal organization and/or differentiation appears to be mediated by different molecular mechanisms, further supporting the strict cellular dependence of CD99 functions. In breast carcinoma MCF-7 cells, CD99 may function as a negative regulator of FAK-mediated tumorigenesis and metastasis by recruiting the protein tyrosine phosphatase SHP2 which dephosphorylates FAK and blocks the positive pro-tumorigenic signaling of CD98/integrins/FAK/RhoA/ROCK . In HodgAs a cell surface molecule, CD99 can be easily targeted by antibodies ,52,92,93The selective expression of CD99 in immune and stromal cells   may have a broad effect on an entire tumor cell population. On the other hand, it may be intriguing to explore the possibility of using exosomes to replace the effects of CD99 re-expression in tumors that have lost CD99 during progression.In immune and tumor cells, CD99 is expressed in both short and long forms. Both isoforms play functional roles, and their differential expression can lead to distinct functional outcomes. Although the conjunction of the distinct functional outcomes via a single receptor might be coincidental, it is tempting to speculate that this finding represents a coordinated mechanism. The discrepant roles of CD99 in different tumors may be due not only to the difference in cancer types but also to the difference in the relative expression levels between the two CD99 isoforms. However, the currently available anti-CD99 antibodies cannot distinguish between the two CD99 isoforms, and other approaches must be considered in evaluating the expression levels of each CD99 isoform separately.The regulation of CD99 expression is still far from being understood. Considering the role that the loss of CD99 has in many common tumors, a true appreciation of the possible mechanisms involved  is urgently required. In addition, more detailed studies are necessary to clarify how and when CD99sh is expressed with respect to the preeminent CD99wt isoform in different tumors.Considering the high dependence of CD99 functions on the cellular context, any therapeutic strategy targeting the molecule must be thoroughly studied in the appropriate context before any further clinical development.Critical issues:The application of chimeric antigen receptor (CAR) T cell therapy is emerging as a very promising, novel strategy, particularly after its success in the treatment of certain hematologic cancers. However, CAR T cell therapy for solid tumors faces some challenges, and its application potential is still far from being fully exploited because of the lack of specific tumor antigens, tumor heterogeneity, and the presence of immunosuppressive factors in the tumor microenvironment ,110. CD9Recent advances in our understanding of cancer have revealed that tumors should be considered as complex tissues in which cancer cells communicate directly and indirectly with the surrounding cellular microenvironment to promote their own survival ,112. BecInteractions between tumor cells and their microenvironment involve extracellular vesicles, which are released by cells under physiological and pathological conditions and contain a bioactive cargo . The presence of CD99 in exosomes may provide another perspective on the role of this molecule in tumorigenesis.New perspectives:CD99 is clearly involved in regulating tumor growth and differentiation and, even more importantly, in cell migration/adhesion and metastasis. In addition, the molecule is expressed in both immune and stromal cells and may have a function in the interplay between tumor cells and normal cells reacting to the tumor . However, the role of CD99 in tumorigenesis has been largely underestimated. The mechanisms of action are complex and strictly cell type-dependent, and a more profound level of understanding is required to fully exploit the therapeutic potential of strategies targeting CD99.


Producing molecular imprinting-based materials has received increasing attention due to recognition selectivity, stability, cast effectiveness, and ease of production in various forms for a wide range of applications. The molecular imprinting technique has a variety of applications in the areas of the food industry, environmental monitoring, and medicine for diverse purposes like sample pretreatment, sensing, and separation/purification. A versatile usage, stability and recognition capabilities also make them perfect candidates for use in forensic sciences. Forensic science is a demanding area and there is a growing interest in molecularly imprinted polymers (MIPs) in this field. In this review, recent molecular imprinting applications in the related areas of forensic sciences are discussed while considering the literature of last two decades. Not only direct forensic applications but also studies of possible forensic value were taken into account like illicit drugs, banned sport drugs, effective toxins and chemical warfare agents in a review of over 100 articles. The literature was classified according to targets, material shapes, production strategies, detection method, and instrumentation. We aimed to summarize the current applications of MIPs in forensic science and put forth a projection of their potential uses as promising alternatives for benchmark competitors. The area of forensic science emerged due to the need for scientific techniques for investigating and proving crimes. Forensic science is quite a multidisciplinary area of study with ten and more subdivisions like chemistry, biology, toxicology, geology, archeology, anthropology, astronomy, engineering, etc. All these subdivisions have different methods for problem solving and use a series of specialized tools. In general, “problem solving” in the area of forensic analysis have two meanings: First, identifying the physical evidence or a questioned sample, and the second one is comparing the results with a known material to find the origin of the questioned sample. Molecular imprinting techniques present solutions for both of these requirements as well. Due to their versatility, molecular imprinted polymers (MIPs) have gained many applications in a variety of areas . VersatiMolecular imprinting is the method of producing tailor-made complementary cavities against a targeted structure called the template. By means of these smart and complementary cavities, the resulting molecules have both chemical and physical recognition capabilities; therefore, they are also classified as biomimetic receptors or plastic antibodies . MoleculIn imprinting history, researchers have integrated several polymerization techniques into the imprinting process which has resulted in five main types of molecular imprinting techniques. These can be summarized as covalent, non-covalent, semi-covalent, ionic, and metal coordination methods . A schemEach of these techniques has certain advantages with respect to the affinity, selectivity, kinetics and reproducibility of the final polymers.In this review, we have focused our attention on compiling the imprinting literature for forensic science applications after briefly summarizing the history of imprinted polymers. In this context, we have clustered the studies into groups related to target structures, material shapes, production strategy, application method and detection platforms. In each subsection, we summarize the related studies while mentioning its novelty, contribution and importance for forensic science applications.Availability of molecular imprinting as an analytical tool for variety of target structures is advantageous on forensic chemistry and forensic toxicology. Some of target molecules related to forensic sciences were summarized below:Legal/illicit Drugs: There is a wide range of abused legal or illicit drugs that come across in forensic cases. Some of the most common drugs are given below with the examples from the molecular imprinting literature. Pain relievers, cough/cold medicines, prescription sedatives like benzodiazepines, and barbiturate sleeping aid medicines are the most studied drugs [r > 0.98) with a low LOQ quantification < 10 μg/L that also had an acceptable precision and accuracy for day-to-day and in-day measurements [ed drugs ,7,8,9. Aed drugs . AndersoCannabinoids like marijuana and hashish are the most commonly used illicit drug ,11,12. N−7 M) at levels of selectivity similar to those of antibodies. They concluded that the high binding affinities and selectivity could lead to the use to MIPS in enzyme- based assays like ELISA and immune-affinity techniques for isolation/separation of water-soluble biologically related compounds [3-folds) in comparison to the competitor molecules [Opioids like heroin and opium are substances that act on the opioid receptors to produce morphine-like effects ,14,15. Aompounds . PiletskSalvia divinorum, and dextromethorphan  were extensively examined targets [Another class of the drugs often studied are stimulants like cocaine, amphetamine, and methamphetamine ,16,17. A targets . Djozan Hallucinogens like lysergic acid diethylamide (LSD), mescaline, psilocybin (magic mushrooms) are another class of targets . Chapuis−8–1.0 × 10−4 M with a detection limit of 2.5 × 10−8 M [−5 M, similar to those of the natural analog (acetylcholine esterase). They concluded that MIP-based artificial receptors are very useful for isolating and separating water-soluble biologically related compounds and detecting nicotine levels for addicted patients [The compounds including anabolic steroids, inhalants (solvents and gases), nicotine, and alcohol are other intensively examined targets ,23,24,25× 10−8 M . Krupadapatients . Zhou etpatients . MatsuguForensic analysis approaches are also applicable for anti-doping purposes to determine doping with performance-enhancing drugs, stimulants, steroids, and corticosteroids ,28,29,30−7–1.2 × 10−5 M, 2.1 × 10−7 M, and 99.5%–103.2%, respectively [−8–1.0 × 10−1 M with a LOD value as 30 nM [Poisons including cyanide, arsenic, nightshade, hemlock, curare, nicotine, caffeine, quinine, atropine, strychnine, and brucine were also intensively evaluated targets in the MIP literature ,32,33,34as 30 nM .DNA: Not only small molecules but also biomacromolecules, especially DNA, are supplying important information for forensic investigations. Although DNA sequences are 99.9% the same in every person, the remaining 0.01% is enough to distinguish one person from another, if you have the enough amount of DNA molecule to test [ to test . Therefo to test ,37,38,39 to test . DiltemiExplosives: Nitrobenzene, dinitrotoluene (DNT), trinitrotoluene (TNT), and cyclotrimethylene-trinitramine (RDX) were extensively studied explosives for criminal investigations [igations ,47,48,49igations   and organic  residues [residues   is an extremely sensitive spectroscopic technique based on molecular vibrations but it has some drawbacks, as any molecule with a similar chemical structure may interfere with the SERS and cause doubtful results. Thus, molecular imprinting provides a good solution to increase the selectivity. Holthoff et al. presented a novel strategy to combine molecular imprinting technology with SERS to improve the detection of explosives via deposing imprinted xerogels which provide a high porosity along with large surface areas on SERS active surface [ surface . By usinIon mobility spectrometry (IMS) is a widely used analytical method used for detection and identification of trace amounts of vapors based on the mobility of gas phase ions in a weak electric field. IMS has many uses in the area of forensic sciences, including personal markers, and detection of explosives, toxic substances and narcotics [arcotics . There harcotics ,73,74.Surface plasmon resonance (SPR) is an optical sensor based on optical excitation of surface electrons of metal on a metal-dielectric interface. SPR is a widely used method due to its availability for mobility, ease of modification and production of sensor surfaces available for multi-analyte detection via spotting and sensitivity [sitivity .Molecular imprinting-based SPR sensors are used for a variety of biological structures like proteins, antibodies and antibody fragments, hormones, cells, viruses, aptamers ,77 as wePhotonic crystals are periodic nanostructures that affect the motion of photons that can also be used as label-free detecting platforms due to their structural coloration ability. Hu et al. demonstrated a method for combining photonic crystals with molecular imprinting technology via hierarchical porous structured molecularly-imprinted hydrogel for sensor applications . They acChemiluminescence sensing is the one of the more sensitive colorimetric sensing strategies. Han et al. developed a molecular imprinting-based electroluminescence sensor which has the possibility of becoming an alternative to a trained dog for the detection of hidden drugs in luggage, mail, vehicles, and aircraft, and in the human body [−15 M was achieved. This method is quite promising for detection of hidden drugs or explosives from their odor. There are also molecular imprinted chemiluminescence sensors for poisons in the literature. Liu et al. developed a molecular imprinted chemiluminescence sensor for the determination of brucine, which is a dangerous poison. The detection limit was reported as 2.0 × 10−9 g/mL [man body . They ac0−9 g/mL .Fluorescence sensing is another colorimetric sensing approach for ultrasensitive target detection in combination with MIPs. Fluorescence emission may be obtained by a variety of fluorophore dyes or different-sized quantum dots. Fluorescent-labeled MIP materials could be used for visualizing the recognition between the imprinted material and the template molecules. Use of fluorescent probes in combination with MIPs has mainly focused on the detection of poisons and vapors of explosives and fire debris analysis [analysis ,85,86,87Piezoelectric (PZ) transduction sensors have many sensing applications for a variety of target structures. Their advantages in gas phase detection are quite important for on-site analysis. Quartz crystal microbalances, surface acoustic wave-guides, microcantilevers, and micro-electromechanical systems are among intensively studied mass sensitive sensor systems.Quartz crystal microbalance (QCM): Chianella et al. reported a molecular complete imprinting solution to detect microcystin-LR, a cyanobacteria-based toxin [ed toxin . They prSurface/bulk acoustic wave: Surface acoustic wave (SAW)- and/or bulk acoustic wave (BAW)-based sensors work according to the variation of velocity of the acoustic waves based on the increased mass on the transducer. Percival et al. studied a molecular imprinting-based surface acoustic wave sensor for the detection of an anabolic steroid, nandrolone. They synthesized MIP layers via a covalent imprinting approach and reported a frequency shift of up to 0.2 ppm with the sensor preferring the target, nandrolone, to analogous compounds. They also concluded that such acoustic wave devices could be integrated into lightweight and low cost oscillator circuits allowing inexpensive screening technique to be developed [3 and after 18 months, its detection signal decreased by about only 4.4% [eveloped . Pan et nly 4.4% .−9–1.0 × 10−4 M at pH 8.0 with a LOD value of 5.0 × 10−9 M and high recoveries between 96.1% and 105.6% [Even though caffeine is a widely used legal natural stimulant, it has also importance for forensic purposes. Tanada et al. reported caffeine analysis can be used for forensic hair discrimination . Liang ed 105.6% .−3 μM.Tan et al. produced a molecular imprinting-based BAW sensor for the detection of paracetamol, a frequently encountered drug in suicide cases . They reMicrocantilevers: Use of atomic force microscopes (AFMs) for chemical and biological sensing is a trending area. A modified AFM microcantilever AFM was used as mechanical transducer via the increased mass on the modified cantilever with binding of complementary species. This method may be seen as an artificial nose due to the availability of odor detection at ultra-trace levels. Implementing microcantilever-based detection with molecular imprinting is a relatively new area of study [of study .Micro-electromechanical systems (MEMS): MEMS are made up of components between 1 and 100 micrometers in size for miniaturized sensors, actuators, and structures. The use of molecular imprinting technology is a relatively new area for this approach as well. Holthoff et al. developed a microbeam-based MEMS gas sensor with molecular imprinting for the detection of TNT and dimethyl methylphosphonate (DMMP), a simulant for the nerve gas sarin [as sarin . To form2 = 0.996. LOD and LOQ values were reported as 26.9 and 81.6 ppb. They also showed very good repeatability with precision values %) of 1.098% and 0.55% for 1108 and 2216 ppb, respectively. They concluded that the sensor can selectively detect TATP in presence of other explosives including pentaerythritol tetranitrate, 1,3,5-trinitroperhydro-1,3,5-triazine, octahydro-1,3,5,7-tetranitro-1,3,5,7-tetrazocine, and 2,4,6-trinitrotoluene [Electrochemical sensors are a main sensor platform due to their simplicity, cost-efficiency and widely usability. Triacetone triperoxide (TATP) is one of the most common components of explosives and therefore Mamo et al. produced a highly sensitive electrochemical sensor for TATP via a differential pulse voltammetry-based molecular imprinted sensor. They produced a MIP-based glassy carbon electrode which demonstrated good performance at low concentrations for a linear and wide concentration range of 82–44,300 parts per billion (ppb) with a high correlation coefficient of Rotoluene .There are many studies on MIP-based chromatographic methods in the area of forensic sciences like determining gunshot residues from hand and clothes. These residues are obtained by application of adhesive tapes to suspected person hands and clothes which may interfere with the results due to the adhesives on the tapes. Pereira et al. produced molecularly imprinted polymers for diphenylamine removal from organic gunshot residues, which is one of the most common components of gunshot residue .Capillary electrophoresis (CE) is a widely used method for trace analysis and it is a valuable tool in the area of forensic chemistry for analyzing inks, dyes, gunshot/explosive residues and drugs. Deng et al. produced a fiber-based SPME method by using molecular imprinting in capillary electrophoresis application for ephedrine and pseudoephedrine. Even though ephedrine and pseudoephedrine are therapeutic drugs, their amphetamine-like effects at high doses makes them important for forensic purposes. Limits of detection values were increased from 0.20 to 0.00096 μg/mL for ephedrine and 0.12 to 0.0011 μg/mL for pseudoephedrine via successful molecular imprinting-based SPME [sed SPME . As mentsed SPME . They apHeat transfer methods are one of the recent approaches to develop selective biosensors in combination of MIPs. Peters et al. presented a novel approach for molecular imprinting-based nicotine detection from saliva via differential heat transfer resistance [2) of 0.97. LOD value reported as 125 nM was proper for biological samples in which nicotine levels vary in the range of 0–500 μM. Due to the importance of detecting small molecules for forensic purposes, this method offers a promising alternative.sistance . With thIn this review, we aimed to draw a projection on the use of molecular imprinting in the related areas of forensic science. There are two main challenges with forensic science applications: (i) the complexity of the analytes and (ii) a quite low concentration of the analyte in this complex medium. Therefore, MIPs-based materials are intensively utilized as pre-concentrators before quantification through commercial techniques. Meanwhile, the integration of MIPs into traditional setups has recently attracted the efforts of the researchers . Online


A) as an example, the relationship between molecular packing and charge-transport parameters has been computed and analysed. The conducting properties of A are also predicted within the framework of hopping models.Taking 4-aniline salicylaldehyde hydrazone ( A) can apparently overcome the dynamic intramolecular rotations and the intramolecular steric repulsion, effectively enhancing the molecular rigidity and decreasing the internal reorganization energy. The conducting properties of A have also been simulated within the framework of hopping models, and the calculation results show that the intrinsic electron mobility in A is much higher than the corresponding intrinsic hole mobility. These theoretical investigations provide guidance for the efficient and targeted control of the molecular packing and charge-transport properties of organic small-molecule semiconductors and conjugated polymeric materials.Based on first-principles calculations, the relationship between molecular packing and charge-transport parameters has been investigated and analysed in detail. It is found that the crystal packing forces in the flexible organic molecule 4-aniline salicylaldehyde hydrazone ( In this work, we selected flexible A as an example to calculate its reorganization energies in the gas phase and in the crystalline state, and to discuss the effect of packing forces on the reorganization energies. The effective electronic couplings between adjacent A molecules are predicted, and the way these couplings are affected by the molecular orbital shape and relative positions of the interacting units is addressed in detail. Based on these quantum-chemistry calculation results, combined with the Marcus–Hush electron-transfer theory, we simulated the anisotropic electron and hole mobilities of A and provide here an assessment of its field-effect properties as a potential p-type, n-type or ambipolar organic semiconducting material.Recently, a new flexible organic molecule, 4-aniline salicylaldehyde hydrazone . As shown in a)Fig. S2 and hole-transfer reorganization energy (λh) should be much larger than the values observed for typical π-conjugation molecules with good rigidity/planarity, such as oligoacenes and oligofurans. Our calculation results show that the λh value is 0.405 eV, which is considerably larger than those of pentacene /B3LYP calculations on the reorganization energies of A molecules are all nearly planar structures : the inter-ring torsion angles lie between 8.6 and 9.9°, suggesting that the dynamic intramolecular rotations of the conjugate plane and the intramolecular steric repulsion in A are apparently overcome by the packing forces in the crystal. Our calculations yielded λh = 0.101 eV and λe = 0.274 eV for a molecule of A in a tightly packed crystal, which are much smaller than the calculation results for the gas phase. In order to evaluate the effect of the dispersion corrections on the reorganization energy results, the functional B3LYP-D3 and basis set 6-311g were also performed for neutral and charged A. The calculation results show that the electron-transfer and hole-transfer reorganization energies at B3LYP-D3/6-311g (λe = 0.273 eV and λh = 0.105 eV) nearly equal the results at B3LYP/6-311g, consistent with our previous studies  and 1d), we can see that the relaxation of the C—C bonds upon electron transfer is similar to that of the same bonds upon hole transfer, while the geometric changes in dihedral angle between the conjugate planes are more pronounced upon electron transfer than upon hole transfer, which is consistent with the observation of larger λe than λh in the gas phase. This suggests that the large difference between λe and λh (0.153 eV) comes mainly from the torsional vibration between the N atom and its neighbouring benzene ring. In the crystal, both the C—C bonds and the dihedral angles undergo geometric changes to a much smaller extent , which indicates that the intramolecular C—C stretching vibrations and intramolecular rotations are greatly restricted in the aggregated state. This well explains the significantly smaller λe and λh values of A in the crystal than in the gas phase. Moreover, it should be noted that the geometric relaxations in the C—C bonds and dihedral angles occur predominantly upon electron transfer; in contrast, the smaller geometric changes upon hole transfer contribute less to λh. Both bond length and dihedral angle variations are responsible for the larger λe than λh in the crystal.In order to analyse qualitatively the contribution of different vibration modes to the variation in reorganization energies, the changes in bond length and dihedral angle upon oxidation and reduction of A molecules. As shown in Fig. 2a), the crystal structure of A is a tight herringbone arrangement, which is quite common in chain molecule crystals. Compound A crystallizes in the monoclinic system in space group C2/c, and each unit cell has 16 molecules . Examination of the geometries of the hopping pairs reveals that there are two types of molecular contact among all the pathways: (i) the two A molecules are parallel to each other (pathways P1 and P2), which are usually defined as P-type dimers; and (ii) the two A molecules deviate significantly from being parallel , which were defined as T-type dimers in our previous studies. In these hopping pathways, intermolecular electronic coupling takes the form ijS is the spatial overlap, ijJ are the charge transfer integrals, and ie and je are site energies  the shape of the frontier molecular orbitals of the isolated molecule of A; and (ii) the overlap pattern of the highest occupied molecular orbital (HOMO) or lowest unoccupied molecular orbital (LUMO) wavefunctions in the dimers, which is determined by the relative positions of the interacting units. As shown in Figs. 3a) and 3b), the HOMO is mainly located on intra-ring C—C bonds and partially on the C=N bond and O atom, which are aligned predominantly along the short molecular axis, whilst the LUMO is mainly located on the C—C bonds and partially on the C—N bond and O atom, which are aligned predominantly along the long axis. These distribution characteristics of the HOMOs and LUMOs lead to different extents of the spatial overlap of the HOMO–HOMO wavefunctions with the spatial overlap of the LUMO–LUMO wavefunctions in the same dimer. In the P dimers, there is a displacement of more than one benzene ring along the short molecular axis , and thus the electronic couplings are mainly decided by the interaction between the frontier molecular orbitals distributed on the edge part of the monomers. In this configuration, the overall extent of the spatial overlap of the LUMO wavefunctions is much greater than that of the HOMO wavefunctions, which well explains the stronger electronic couplings for electron transfer than that for hole transfer. Similarly, the T dimers show a typical face-to-edge packing mode , and the LUMOs distributed on the edge part of the monomer are responsible for the strong electronic couplings for electron transfer.The calculated electronic couplings are collected in Table 1A using the mobility orientation function proposed by Han and co-workers . From the simulation results we can conclude that: (i) as a molecular crystal A has the potential to be developed as a potential n-type or ambipolar organic semiconducting material, due to the fact that the optimum values of electron-transfer mobility are larger by nearly one order of magnitude than the corresponding optimum values of hole-transfer mobility; and (ii) the maximum predicted hole-transfer and electron-transfer mobilities are present in the same direction, which means that the hole and electron mobilities could achieve their highest values at the same time if the transistor channel orientation were in the direction of Φ = 80°/170° .Comparing these results, it is easy to find that the intermolecular electronic couplings are nearly one order of magnitude less than the corresponding λ. In this case, the hopping model might be preferred for the prediction of carrier mobilities. Here, we simulated the anisotropic hole and electron mobilities for 3.A not only maximizes intermolecular interactions, but also enhances the molecular rigidity and decreases the internal reorganization energy effectively. Furthermore, the conducting properties of A were also simulated within the framework of hopping models, and our calculation results show that the intrinsic electron mobility in A is much higher than the corresponding intrinsic hole mobility, which means that A may be a better electron transporter than hole transporter.We have shown that the most crucial parameters affecting charge-transport properties, the charge-transfer integral and the reorganization energy, are sensitive to the molecular packing, and a densely packed crystal of the flexible π-conjugated compound Our theoretical investigations here are helpful for understanding the relationship between crystal structure and intermolecular charge-transport behaviours, and provide guidance for the efficient and targeted control of the molecular packing and charge-transport properties of organic small-molecule semiconductors and conjugated polymeric materials.10.1107/S2052252517010685/lt5007sup1.pdfSupporting information file. DOI:


Aberrant regulation of dopamine system function is thought to contribute to psychosis in schizophrenia patients; however, the brain regions associated with this dysregulation have not been conclusively demonstrated. We have recently demonstrated that medium spiny neurons in the nucleus accumbens (NAc) receive convergent input from the ventral hippocampus (vHipp) and paraventricular nucleus of the thalamus (PVT). Furthermore, inactivation of either the vHipp or PVT is sufficient to reverse aberrant dopamine system function in rodent models of schizophrenia. Using, chemogenetic experiments we now provide conclusive evidence that the thalamic input to the NAc plays a role in the regulation of dopamine neuron activity. These data demonstrate that the vHipp and thalamus  work in concert to regulate VTA dopamine neuron population activity. Such data are important as they provide evidence that thalamic abnormalities may contribute to the aberrant dopamine system function observed in schizophrenia and suggest that the PVT may be a novel site for intervention in psychosis. To examine this, we explored the orexin system, which is known to provide a dense innervation of the PVT.Pregnant Sprague Dawley (SD) rats were treated on gestational day (GD) 17 with either methylazoxymethanol acetate  or saline. For Poly I:C, pregnant dams were treated on GD12 . Male pups weaned on post-natal day 21 in groups of 2–3 until adulthood (>60 days). For chemogenetic experiments, normal SD rats were bilaterally micro-injected with AAV2 vectors (Addgene) expressing hm3D(Gq)(pAAV-h8yn-HA-hm3D(Gq)-mcherry; 0.5μL) into the PVT or mPFC. Control rats were administered the viral vector lacking the hm3D encoding gene. Prior to testing, CNO  was injected into the nucleus accumbens. In vivo extracellular recordings were performed to measure dopamine neuron activity in the VTA. Spontaneously active VTA dopamine neurons were recorded using previously established electrophysiological criteria.NMDA activation of the PVT induces a significant increase in VTA dopamine neuron population activity. MAM- and Poly I:C-treated rats (both verified rodent models of schizophrenia) consistently display aberrant VTA dopamine neuron population activity, which is restored by pharmacological inactivation of the PVT with TTX. Chemogenetic activation of PVT neurons projecting to the mPFC do not affect VTA dopamine neuron activity; however, activation of PVT neurons projecting to the nucleus accumbens induces a significant increase in dopamine neuron population activity. This effect can be replicated in rats that receive microinjections of the endogenous orexin peptide A or B into the PVT. Consequently, dopamine neuron function can be restored in MAM-treated rats that received a systemic injection of the orexin peptide antagonist TCS 1102.We now demonstrate that orexin receptors are expressed on PVT neurons projecting to the NAc and may serve as a substrate for pharmacological manipulation of this pathway. Here, we provide evidence that both systemic and intracranial (PVT) administration of the orexin receptor antagonist, TCS1102, can normalize aberrant dopamine system function in a rodent model of schizophrenia. Collectively, these data suggest that targeting orexin signaling in the thalamus, specifically, the PVT, may represent a novel site of intervention for psychosis associated with schizophrenia.


The patient successfully underwent robotic assisted radical left adrenalectomy and was diagnosed with a dopamine secreting pheochromocytoma. Pathology revealed increased malignant potential associated with the tumor. The patient underwent full metastatic workup, which was negative. At two years of follow-up there was no recurrence and normalization of lab values.We report on a 43-year-old, asymptomatic female who presented with incidental finding of left adrenal mass. MRI gave concerns for possible pheochromocytoma but markers for pheochromocytoma were not elevated as expected. 24-hour urine dopamine levels (6988  Pheochromocytomas or paragangliomas are neuroendocrine tumors that arise from chromaffin cells. The annual incidence of pheochromocytoma/paragangliomas is approximately 0.8/100,000 person-years  but may A 43-year-old Hispanic woman was referred to our urology clinic at our institution for an incidental finding of left sided adrenal mass on computed tomography (CT) done for right hip and abdominal pain. CT revealed a 5.8 cm enhancing mass from the left adrenal gland. Confirmatory MRI revealed a 6 cm left adrenal mass . The patμg/day (0–250). Surgical intervention was recommended to the patient likely for a dopamine producing pheochromocytoma (dopaminoma). The patient elected to proceed with robotic assisted left radical adrenalectomy. Preoperatively patient was placed on alpha blockade with phenoxybenzamine. Intraoperative and postoperatively patient had no issues with blood pressure control. There were no postoperative complications.Patient was referred to endocrinology for further workup for possible pheochromocytoma. 24-hour urine metanephrines 220 mcmol/mol (0–172) and normetanephrines 276 mcmol/mol (0–247), as well as plasma normetanephrines 1.15 nmol/L (0.00–0.89) and metanephrines .53 nmol/L (0.00–0.49), were all mildly elevated. Urine epinephrine and norepinephrine were normal. 24-hour urine dopamine was drawn due to minimal elevation of metanephrines and normal catecholamines. Dopamine levels were severely elevated, >25x normal range at 6988 Gross pathology revealed 4 × 4 × 3 cm black partially encapsulated mass attached to adrenal gland. Tumor was revealed to be a pheochromocytoma with staining pattern positive for chromogranin, synaptophysin, and neuron specific enolase supporting the diagnosis of pheochromocytoma. The tumor had focal extension through capsule into surrounding adipose tissue along with focal vascular invasion. Tumor was assigned a Pheochromocytoma of the Adrenal Gland Scaled Score (PASS) score of 4  increasing risk of malignant potential. The mitoses were less than three per high power fields, and no atypical mitotic figures were noted. The patient was diagnosed with a dopamine secreting pheochromocytoma.μg/day preoperatively to 180 μg/day (0–250) postoperatively. Normetanephrines and metanephrines did not have a significant change. All labs remained within normal levels at follow-up two years postoperatively. Due to PASS score of 4, (PET) scans were obtained at 6 months and two years postoperatively with no signs of disease. Metaiodobenzylguanidine (MIBG) scan was also done postoperatively with no uptake noted. Genetic testing for multiple endocrine neoplasia type 2, neurofibromatosis type 1, and von Hippel Lindau were negative.After mass resection, 24-hour plasma and urine dopamine, normetanephrines, and metanephrines were repeated at 1 and 6 months and were normal. Urinary dopamine decreased from 6988 Dopamine secreting pheochromocytomas are extremely rare, as only 35 abdominal dopaminomas have been reported in the literature in 35 years. . PoirierAnother issue with dopamine secreting pheochromocytomas is difficulty in diagnosis. Many institutions do not routinely screen for dopamine when doing urinary or plasma catecholamine screening , 4. Thisα- and intraoperative β-blockade to prevent life-threatening hypertensive crises. Alpha-blocker treatment has been considered contraindicated for dopamine secreting pheochromocytomas because case reports have shown it can lead to cardiovascular collapse and potential hypotensive crisis [α-receptors but acts on D1 and D2 receptors. In our case urinary dopamine was used as a surrogate for plasma dopamine preoperatively due to extreme elevation of urinary dopamine. Urinary dopamine is not a direct correlation to circulating plasma dopamine but previous studies looking at dopamine secreting paragangliomas have shown urinary outputs of dopamine were positively related to plasma concentrations of dopamine [Historical management for a classic pheochromocytoma is preoperative e crisis . This ace crisis . This dedopamine . Our patdopamine  have alsdopamine . The resDopamine secreting pheochromocytomas are rare. These tumors have increased risk of malignant potential and may be difficult to diagnose due to atypical symptoms and lack of routine screening for dopamine. Robotic assisted adrenalectomy is an acceptable method of tumor excision in appropriately selected patients.


Staphylococcus aureus and are the most common causative agent in staphylococcal food poisoning. The staphylococcal enterotoxin A (SEA) has been associated with large staphylococcal food poisoning outbreaks, but newer identified SEs, like staphylococcal enterotoxin H (SEH) has recently been shown to be present at similar levels as SEA in food poisoning outbreaks. Thus, we set out to investigate the thermo-stability of the three-dimensional structures of SEA, SEH and staphylococcal enterotoxin E (SEE), since heat inactivation is a common method to inactivate toxins during food processing. Interestingly, the investigated toxins behaved distinctly different upon heating. SEA and SEE were more stable at slightly acidic pH values, while SEH adopted an extremely stable structure at neutral pH, with almost no effects on secondary structural elements upon heating to 95°C, and with reversible formation of tertiary structure upon subsequent cooling to room temperature. Taken together, the data suggests that the family of staphylococcal enterotoxins have different ability to withstand heat, and thus the exact profile of heat inactivation for all SEs causing food poisoning needs to be considered to improve food safety.The staphylococcal enterotoxins (SEs) are secreted by the bacteria  Staphylococcus aureus secretes a wide variety in virulence factors like the staphylococcal superantigen  × mdeg cm2 dmol−1) based on 233 amino acid residues and a molecular weight of 27.1 kDa for SEA and 26.8 kDa for SEE, respectively, and 217 amino acids and a molecular weight of 25.1 kDa for SEH. Protein concentrations used were 0.2 mg/ml, determined by UV/Vis absorbance spectroscopy using the extinction coefficient at 280 nm of 37945 M-1 cm-1 for SEA, 36455 M-1 cm-1 for SEE and 26485 M-1 cm-1 for SEH. Deconvolution of the CD spectra into pure component spectra was performed using the CD secondary structure analysis algorithm BeStSEl with the data range from 200 to 250 nm  for each spectrum measured in millidegrees (mdeg). Protein concentrations used were 0.2 mg/ml in 20 mM NaH2PO4 at pH 5.0 supplemented with either 1 mM EDTA or 0.1 mM ZnCl2.Far-UV CD spectra recorded on SEA at pH 5.0 in the presence of ((DOCX)Click here for additional data file.


Rewards are fundamental to everyday life. They confer pleasure, support learning, and mediate decisions. Dopamine-releasing neurons in the midbrain are critical for reward processing. These neurons receive input from more than 30 brain areas and send widespread projections to the basal ganglia and frontal cortex. Their phasic responses are tuned to rewards. Specifically, dopamine signals code reward prediction error, the difference between received and predicted rewards. Decades of research in awake, behaving non-human primates (NHP), have shown the importance of these neural signals for learning and decision making. In this review, we will provide an overview of the bedrock findings that support the reward prediction error hypothesis and examine evidence that this signal plays a role in learning and decision making. In addition, we will highlight some of the conceptual challenges in dopamine neurophysiology and identify future areas of research to address these challenges. Keeping with the theme of this special issue, we will focus on the role of NHP studies in understanding dopamine neurophysiology and make the argument that primate models are essential to this line of research. Rewards are a central feature of everyday life that promote learning, incentivize value-based decisions, and provide a currency for social exchanges. Dopamine neurons respond to rewarding events. Specifically, phasic dopamine activity reflects the difference between received and predicted rewards, i.e., reward prediction errors. Systems neuroscience studies in non-human primates (NHP) have been critical to understanding this coding scheme, as well as investigating the behavioral implications of reward prediction error coding.Right now, we stand at the threshold of a new neuroscientific age. This age will be defined by big data and molecular control of neural information processing. From this vantage point, this review endeavors to provide a brief introduction to dopamine neurons, examine the critical findings revealed by NHP studies that have shaped our understanding of dopamine function, and evaluate what role this valuable species should have in future investigations. We will attempt to identify outstanding challenges to our current understanding of this critical brain system, propose areas for future research, and reinforce the need to preserve and technologically advance behavioral neurophysiology studies in NHP. Keeping with the theme of this special issue on NHP studies of basal ganglia function, this review does not attempt to be comprehensive and passes over many of the recent advances observed in rodent models. In addition, we do not discuss the role of dopamine cell loss in Parkinson’s disease, nor the valuable contribution of the monkey MPTP Parkinson’s disease model , and ventral tegmental area (VTA), respectively. . l-DOPA is converted to dopamine via aromatic l-amino acid decarboxylase (AAAD) and packaged into vesicles by the vesicular monoamine transporter (VMAT)  tonic activity and phasic (burst firing) activity when the impulse rate can briefly reach 20–30 imp/s , whereas rewards that are worse than predicted inhibit dopamine activity (negative prediction error responses). Modeling studies demonstrated that the prediction error term in popular reinforcement learning (RL) algorithms closely resembles the phasic dopamine signal  as a function of physical value Debreu . Risk-avAn outstanding question related to economic value coding is whether economic costs reduce the magnitude of dopamine responses. In one study, increasing the effort required to get a reward resulted in reduced responses of some dopamine neurons, but not others  determining whether dopamine neurons are functionally homogenous, (2) elucidating the relationship between behavioral measures and striatal dopamine release, and (3) defining the functional role of dopamine signals in the cortex.Several studies have reported that a fraction of dopamine neurons are excited by novel or aversive stimuli and outcomes (Brischoux et al. Multiple aspects of dopamine signaling may contribute to the observed complexity of dopamine responses, including context dependency and complex temporal dynamics. Dopamine neurons are exquisitely sensitive to the experimental context. For instance, when visual stimuli predict both appetitive and aversive stimuli, approximately 40% of dopamine neurons respond to the stimulus predicting the aversive outcome (Matsumoto and Hikosaka The temporal dynamics of the dopamine response can further complicate the interpretation of these responses. Short latency activations can occur 50–90 ms following behavioral events that reflect physical impact, novelty, and stimulus generalization Fiorillo . These sClassic studies have repeatedly shown that, even during operant paradigms, dopamine responses are time-locked to external reward predictors, rather than to the onset of well controlled, single joint movements or associated EMG activity Fig. a Ljungb. In contIt is widely believed that phasic dopamine signals update action values in the striatum (Shen et al. Electrophysiological recordings from dopamine cell bodies in the midbrain have demonstrated that phasic dopamine responses code reward prediction error, the difference between received and predicted reward. Studies in awake, behaving NHP have been critical to this endeavor, because they are highly trainable and can provide a wealth of data through in-depth exploration of single unit dopamine activity. NHP possess a rich and complex behavioral repertoire which has led to advanced understanding of the role of dopamine in learning, movement, and decision making. Not discussed here but worth mentioning, the MPTP monkey model has been critical to the study of neuronal and behavioral deficits associated with Parkinson’s disease. In short, the unique properties of NHP models have made them essential to understanding midbrain dopamine function and dysfunction.The findings reviewed here demonstrate that, even for studying a relatively simple and evolutionarily old neural structure like dopamine neurons, there are significant advantages to using NHP models. Non-human primates possess behavioral and anatomical characteristics that are more similar to humans than any other experimental animal model. From a behavioral standpoint, the cognitive capability and choice flexibility reviewed here and demonstrated elsewhere (Eiselt and Nieder A new generation of molecular tools—including optogenetics and in vivo single cell imaging—has revolutionized how we ask questions and even what questions we can ask. These technologies, however, have not been widely incorporated into monkey neurophysiology studies. Although progress is being made, as reviewed elsewhere in this issue (Galvan et al.