Contingency Training Alters Neurobiological Components of Emotional Resilience in Male and Female Rats.

Prior research with a rat model of behavioral therapy [i.e., effort-based reward (EBR) contingency training] suggests that strengthened associations between physical effort and desired outcomes enhance neurobiological indices of resilience. In the current study, male and female Long-Evans rats were exposed to either six weeks of EBR training or noncontingent training prior to 10 days of exposure to chronic unpredictable stress (CUS). Subsequently, all animals were exposed to a problem-solving task and then trained in a spatial learning/foraging task, the Dry Land Maze (DLM). Following habituation training and test trials, rats were assessed in a probe trial that generated a prediction error (cognitive uncertainty). Results indicated that, during CUS exposure, contingency-training enhanced dehydroepiandrosterone/corticosterone ratios (consistent with healthier stress responses), especially in male rats. Additionally, contingency training increased exploratory behaviors in the probe trial as well as differentially influenced on-task problem-solving performance in males and females. Following the probe trial, brains were exposed to histological analyses to determine the effects of sex and contingency training on various neurobiological markers. Contingency training decreased BDNF-immunoreactivity (ir) in the hippocampus CA1 and lateral habenula, implicating differential neuroplasticity responses in the training groups. Further, coordinated fos-ir activation in areas associated with emotional resilience (i.e., motivation-regulation) was observed in contingent-trained animals. In sum, the current findings confirm that behavioral training is associated with neurobiological markers of emotional resilience; however, further assessments are necessary to more accurately determine the therapeutic potential for the EBR contingency training model.

On the needle clogging of staked-in-needle pre-filled syringes: Mechanism of liquid entering the needle and solidification process.

Staked-in-needle prefilled syringes (SIN-PFS) are widely used for the parenteral administration of drug product solutions. During stability studies, clogging of the injection needle was observed in syringes filled with concentrated antibody solution. A prerequisite for this phenomenon is that liquid has entered the needle. In this study, we characterized the mechanism causing the entry and movement of liquid in the needle using neutron imaging without manipulating the container closure integrity of the syringe. The gas pressure difference between inside and outside of the syringe was identified as the major cause of liquid movement. The influence of external factors, such as temperature fluctuation and physical pressure on the stopper, were tested and were confirmed to have a relevant impact on the processes of liquid entering and moving inside the injection needle. In a second step, the solidification process of the liquid segments inside the needle via solvent evaporation was further investigated, and the process was found to be dependent on storage time, environmental climate and interaction between the drug product solution and the needle surface. The presence of air/liquid segments was identified as a further factor for the stochastic behavior of needle clogging. For the first time, this fundamental mechanism behind the needle clogging issue was investigated in depth and the results will help to reduce the defect rate for clogged SIN-PFS products.

Clogging in staked-in needle pre-filled syringes (SIN-PFS): Influence of water vapor transmission through the needle shield.

Staked-in needle pre-fillable syringes (SIN-PFS) are a convenient delivery system widely established in the growing pharmaceutical market. Under specific storage conditions, the needle of PFS containing high concentration drug product (DP) solution is prone to clogging, which prevents administration of the liquid. The purpose of this study is to clarify the clogging phenomenon of SIN-PFS and to elucidate the role of water vapor transmission via the needle shield. The presence of liquid within needles is a prerequisite condition for clogging and was investigated non-invasively by neutron imaging (NI) to confirm that liquid can migrate into the needle under certain processing conditions. The water vapor transmission rate (WVTR) of different needle shields was measured and the impact of temperature and relative humidity (rH) on the WVTR was investigated on sheets with the same composition as used in commercial needle shields. Our study clearly showed that the partial vapor pressure difference (ΔPP) across the needle shield is the dominant driving factor for water vapor transmission. A linear correlation between ΔPP and WVTR was found and a model to predict the water vapor transmission for PFS under specific storage conditions was developed. The impact of the WVTR on needle clogging was confirmed by clogging tests performed on SIN-PFS stored under different conditions. Thereby, we clearly show that high water loss induced by higher WVTR can be correlated to an increased occurrence of needle clogging. In conclusion, the WVTR of the needle shield plays a key role in needle clogging and the established WVTR model can be employed to assess the clogging risk for product development.

Autophagy regulates satellite cell ability to regenerate normal and dystrophic muscles.

Autophagy is emerging as a key regulatory process during skeletal muscle development, regeneration and homeostasis, and deregulated autophagy has been implicated in muscular disorders and age-related muscle decline. We have monitored autophagy in muscles of mdx mice and human Duchenne muscular dystrophy (DMD) patients at different stages of disease. Our data show that autophagy is activated during the early, compensatory regenerative stages of DMD. A progressive reduction was observed during mdx disease progression, in coincidence with the functional exhaustion of satellite cell-mediated regeneration and accumulation of fibrosis. Moreover, pharmacological manipulation of autophagy can influence disease progression in mdx mice. Of note, studies performed in regenerating muscles of wild-type mice revealed an essential role of autophagy in the activation of satellite cells upon muscle injury. These results support the notion that regeneration-associated autophagy contributes to the early compensatory stage of DMD progression, and interventions that extend activation of autophagy might be beneficial in the treatment of DMD. Thus, autophagy could be a 'disease modifier' targeted by interventions aimed to promote regeneration and delay disease progression in DMD.

Paper or Plastic? Exploring the Effects of Natural Enrichment on Behavioural and Neuroendocrine Responses in Long-Evans Rats.

Enriched environments are beneficial to neurobiological development; specifically, rodents exposed to complex, rather than standard laboratory, environments exhibit evidence of neuroplasticity and enhanced cognitive performance. In the present study, the nature of elements placed in the complex environment was investigated. Accordingly, rats (n = 8 per group) were housed either in a natural environment characterised by stimuli such as dirt and rocks, an artificial environment characterised by plastic toys and synthetic nesting materials, a natural/artificial environment characterised by a combination of artificial and natural stimuli or a laboratory standard environment characterised by no enrichment stimuli. Following exposure to emotional and cognitive behavioural tasks, including a cricket hunting task, a novel object preference task and a forced swim task, brains were processed for glial fibrillary acidic protein (GFAP)-, neuronal nuclei (NeuN)- and brain-derived neurotrophic factor (BDNF) immunoreactivity. Baseline and stress foecal samples were collected to assess corticosterone (CORT) and dehydroepiandrosterone (DHEA). Natural environment animals exhibited shorter diving latencies and increased diving frequencies in the second forced swimming task, along with higher DHEA/CORT ratios, and higher GFAP immunoreactivity in the hippocampus. The type of environmental enrichment did not influence levels of BDNF immunoreactivity in the CA1, CA3 and dentate gyrus of the hippocampus; however, natural environment animals exhibited higher levels of NeuN immunoreactivity in the retrosplenial cortex, an area involved in spatial memory and other cognitive functions. These results suggest that, in addition to enhancing behavioural and endocrinological variables associated with resilience, exposure to natural stimuli might alter plasticity in brain areas associated with cortical processing and learning.

FAS-ligand regulates differential activation-induced cell death of human T-helper 1 and 17 cells in healthy donors and multiple sclerosis patients.

Functionally distinct T-helper (Th) subsets orchestrate immune responses. Maintenance of homeostasis through the tight control of inflammatory Th cells is crucial to avoid autoimmune inflammation. Activation-Induced Cell Death (AICD) regulates homeostasis of T cells, and it has never been investigated in human Th cells. We generated stable clones of inflammatory Th subsets involved in autoimmune diseases, such as Th1, Th17 and Th1/17 cells, from healthy donors (HD) and multiple sclerosis (MS) patients and we measured AICD. We find that human Th1 cells are sensitive, whereas Th17 and Th1/17 are resistant, to AICD. In particular, Th1 cells express high level of FAS-ligand (FASL), which interacts with FAS and leads to caspases' cleavage and ultimately to cell death. In contrast, low FASL expression in Th17 and Th1/17 cells blunts caspase 8 activation and thus reduces cell death. Interestingly, Th cells obtained from healthy individuals and MS patients behave similarly, suggesting that this mechanism could explain the persistence of inflammatory IL-17-producing cells in autoimmune diseases, such as MS, where their generation is particularly substantial.

Modeling paternal attentiveness: distressed pups evoke differential neurobiological and behavioral responses in paternal and nonpaternal mice.

With the exception of parturition and lactation, male California deer mice (Peromyscus californicus) exhibit the same parental responses toward offspring as conspecific females. A closely related species, Peromyscus maniculatus, however, rarely exhibits paternal responses. In the current study, a comparative species approach was used to assess paternal responses in both Peromyscus species with varying levels of paternal experience (biological fathers, pup-exposed virgins, and pup-naïve virgins). Of special interest was the persistence of the males to direct their attention toward a distressed pup housed in a small enclosure (i.e., a barrier existed between males and pups). In addition to pup-directed responses, non-pup-directed responses such as grooming, resting and jumping were recorded. Subsequently, all animals' brains were assessed for fos-immunoreactivity (ir) in several areas previously associated with the paternal brain circuit. Overall, P. californicus exhibited more pup-directed responses as well as less fos-ir in brain areas involved in emotional integration and processing such as the insula and anterior cingulate. In addition to increased activation of emotional regulatory areas, P. maniculatus males, observed to direct their behavior away from the pup, exhibited higher fos-ir in the nucleus accumbens (involved in goal acquisition), perhaps due to a heightened motivation to avoid the pups. Interestingly, experience with pups altered the lateral septum and amygdala activation of P. maniculatus to levels similar to P. californicus biological fathers. Finally, fos-ir was increased in the medial preoptic area, involved in the maintenance of maternal behavior, in the biological fathers of both species. Thus, although biological predispositions toward pup-directed behaviors were observed in P. californicus males, evidence of a few shifts toward the paternal neural activation profile was apparent in P. maniculatus males. Specifically, modifications in fear responses and social processing may represent the cornerstones of the gradual shift from social tentativeness to social attentiveness in the presence of pups.

Cytogenetic and molecular cytogenetic profile of bone marrow-derived mesenchymal stromal cells in chronic and acute lymphoproliferative disorders.

The possibility that human mesenchymal stromal cells (hMSC) may derive from the malignant clone in hematological malignancies (HM) is a controversial issue. In order to clarify hMSC origin and disclose possible cytogenetic heterogeneity in hMSC belonging to different patients, bone marrow (BM)-derived hMSC samples from chronic lymphocytic leukemia (CLL) and acute lymphoblastic leukemia (ALL) were expanded in culture, characterized by flow cytometry, and screened by conventional cytogenetic analysis and fluorescent in situ hybridization for the presence of possible cytogenetic aberrations, related or not to the hematopoietic neoplastic clone. Our data showed that the presence of cytogenetic aberrations in successfully expanded HM-MSC stromal layers derives from the persistence of contaminating hemopoietic cells (HC), which is greatly supported by in vitro culture conditions that could mimic in vivo microenvironmental niche. Interestingly, the presence of binucleated HM-MSC maintaining a diploid numerical setting has been also detected, while aneuploidies were observed more frequently in mononucleated HM-MSC from patients with an altered karyotype than in patients with a normal karyotype and controls. In conclusion, here, we showed that in ALL and in CLL, the BM-MSC has a normal karyotype, thus supporting a distinct origin from hematopoietic cells (HC). The presence of in vitro hMSC aneuploidy is associated with lymphoid neoplasias carrying chromosome abnormalities, suggesting that hMSC should be characterized before clinical application. The adequacy of hMSC cytogenetic characterization here proposed could represent a "prerequisite" to standardize the hMSC analysis before their use in the autologous setting and cellular therapy.

Fatherhood alters behavioural and neural responsiveness in a spatial task.

The hormones and experiences of pregnancy, parturition and lactation have been shown to dramatically remodel the female rat's hippocampus, potentially enhancing behaviours critical for meeting the increased demands of motherhood. Previous work in our laboratory has also suggested that pup exposure, apart from pregnancy and lactation, constitutes an important influence on ancillary maternal behaviour (e.g. foraging behaviour). In the present study, we press the parental model further by examining the effect of pup exposure on the hippocampus of males from a biparental mouse species, the California mice (Peromyscus californicus). Males were either Fathers (i.e. first-time fathers housed with a female from mating until 7 days after parturition), pup-exposed virgins (PEV; i.e. sexually naïve males briefly exposed to pups daily for 7 days) or Virgins (i.e. never exposed to females or pups). A dry-land maze (DLM), as used for assessing spatial learning, was employed to determine the foraging abilities of the males. The results indicated that, on the most challenging day of testing (i.e. acquisition day), California mouse Fathers demonstrated superior memory for the task compared to PEVs and Virgins. In addition to the behavioural data, significantly more fos-immunoreactivity was observed in the CA1, CA3 and dentate gyrus regions of the hippocampi of Fathers than PEVs or Virgins in response to the probe trial. Additionally, a trend for altered performance on the DLM was observed in the PEVs on the last day of testing, which was accompanied by the highest levels of nestin-immunoreactivity, an indicant of neuroplasticity, of the three groups. In summary, these data suggest that, in accordance with previous observations of maternal rats, the paternal brain is similarly influenced by parental experience, as demonstrated by accompanying modifications to relevant neurobiological and behavioural responses.

Behavioral and physiological correlates of stress related to examination performance in college chemistry students.

This study was designed to assess physiological and behavioral correlates of academic stress during a college course in organic chemistry in the USA. Participants (45 females, 46 males, mean age 19.88 years) were screened for their basal hypothalamic-pituitary-adrenocortical activity using saliva samples collected at the beginning of the course and after each major test. Displacement activities (DAs) were observed during each test by videotaping students' behavior when they were taking the tests. These variables were then used as predictors of the students' achievement as measured by their grade point average (GPA) scores, American College Testing (ACT) scores, and their final grade in the class. Ninety-one students, enrolled in Organic Chemistry I at Marshall University during the summer of 2009, were recruited for this study. It was found that individual differences in the physiological stress responses are a factor in predicting the students' ability to pass a challenging class. A logistic model built on GPA, DAs during stress, and salivary hormone (cortisol and dehydroepiandrosterone) concentrations was able to correctly classify almost 90% of the students passing the class. The same model was not nearly as successful in determining the possible factors behind failing the class, because the classification success was just 52%, a figure close to chance. We conclude that a clear set of characteristics related to the students' ability and resilience to psychological stress are necessary to succeed in a challenging class. The reason behind dropping or failing a class could be less defined. These data indicated that investigating the physiological and behavioral propensities associated with psychological stress can help us better understand an individual's coping responses to a long-term challenging situation.

Maternal care patterns and behavioral development of rhesus macaque abused infants in the first 6 months of life.

We investigated the maternal care patterns of rhesus macaque mothers who physically abuse their infants, and compared their infants' behavior to that of nonabused infants. Parametric and multidimensional scaling analyses indicated that abusive mothers have a distinct parenting style characterized by high rates of rejection and contact-breaking from their infants. Compared to control infants, abused infants exhibited signs of delayed independence from their mothers including higher rates of distress calls and anxiety, lower rates of contact-breaking, and differences in play. Several aspects of the abused infants' behavior were correlated with rates of abuse received during the first month, or with other maternal behaviors. These findings provide a more comprehensive characterization of the parenting styles of abusive mothers and the early behavioral development of their infants than previously available. Detailed knowledge of the early experience of abused infants is crucial for understanding possible pathological alterations in behavior and neuroendocrine function later in life.

A survey on parent-child conflict resolution: intrafamily violence in Italy.

OBJECTIVE: The purpose of this paper was to investigate factors associated with intrafamily violence toward children in Italy. Family structure and the characteristics of both caretakers and children were explored. Their association with the incidence of minor and severe violence was analyzed to test the hypothesis that child physical abuse is related to a combination of different factors involving the family as a whole. METHOD: This research was carried out by submitting an anonymous questionnaire to 2388 families residing in Tuscany, Italy. The form included two sections, one related to the family cultural substrate, and to potentially influential events affecting the family during the year 1998. The other one, taken from the conflict tactics (CT) scales, presented a hierarchy of possible responses to conflict. RESULTS: Physical punishment appeared to be a general behavior in Italy, because the incidence of minor violence was 77%. The incidence of severe violence was about 8%. When considering family, caretaker, and child characteristics mostly associated with physical punishment in Italy, we found that families with a low income, where caretakers had health problems or were stressed, and with younger and more "problematic" children presented the highest risk of intrafamily violence during conflict resolution. CONCLUSIONS: The results point to the importance of a balance between potentiating and compensatory factors in complex and dynamic relationships among family members, to avoid strategy resolution of conflict ultimately leading to violence. Moreover, minor and severe violence appeared to be two related but different phenomena.

Parental failure in captive cotton-top tamarins (Saguinus Oedipus).

Several New World monkey species experience high rates of infant mortality in captivity, and parental failure in the form of infant neglect and abuse is often regarded as one of the leading causes of this problem. We explored a large archival database to assess environmental, familial, and biological variables identified as significant for parental success in previous studies of captive tamarins, through several generations and across several dozen pedigrees. Using a stepwise multiple regression analysis we developed a model including the fewest variables able to identify statistically significant predictors of infant outcome. We found that seven independent variables could predict infant outcome in the colony. The most important appeared to be the presence of helpers with whom parents could share infant carrying. Mother's experience and litter size were two other variables that contributed to a significant extent to explaining parental failure. Moreover, the model showed that there is a measurable contribution to infant outcome due to the health status of both parents. Finally, we found a distinct role for mothers and fathers, and that parental failure follows different patterns for abuse and rejection.

Social behavior and hormonal correlates during the perinatal period in Japanese macaques.

This work assessed the changes in both social interactions and estrogen metabolite excreted in feces in eight group-living Japanese macaques (Macaca fuscata). We tested the hypothesis that the social behavior of pregnant females shows significant changes during the late prepartum and early postpartum period. We also tested the hypothesis that the marked fluctuation in estrogen levels during the perinatal period is associated with the changes in social behavior. We found that pregnant females withdrew from the social life of their group in preparation for parturition and only slowly regained their normal social activity after delivery. These changes were correlated with the fluctuation in estrogen conjugate excreted in feces, giving further evidence that hormones can enhance responsiveness to the infant and may predict maternal competence in macaques. We also found that the high frequency of self-grooming by pregnant females during the perinatal period may be a functional way to improve the quality of care toward an infant by a simple shifting from the care for oneself to the care for the infant after parturition.

Endometrioid carcinoma in pelvic endometriosis in a postmenopausal woman with tamoxifen adjuvant therapy for breast cancer. A case report.

This concerns a 57 year old woman operated on in 1988 for a left radical mastectomy due to ductal breast carcinoma and subsequently treated with chemotherapy and Tamoxifen adjuvant. In 1990 a laparo-hystero-oophorectomy was carried out due to uterine fibromas. The woman continued taking Tamoxifen. Two years later a pelvic regeneration appeared, resulting in endometriosis, site of adenomatose hyperplasia and of endometrioid carcinoma GI. This furthermore confirms the importance of a gynecological follow-up for all women treated with Tamoxifen adjuvant therapy.

[Danazol in the therapy of dysfunctional uterine hemorrhage in women of fertility age]. / II Danazol nella terapia delle emorragie uterine disfunzionali nella donna in età fertile.

The effectiveness of Danazol in a dose of 200 mg/day for 3 consecutive months has been assessed in 30 women complaining of dysfunctional uterine haemorrhages. The patients were selected by excluding any associated genital pathology (myomas, ovarian neoformations, endometrial cancer, etc.) and any clotting pathology. Danazol proved effective in all cases treated. Side-effects were inconsequential save for 2 women who had to suspend treatment following increase in transaminases in one case and defluvium capitis in the other. The treatment remained effective for at least 2 months after termination. In 26% of the women, the haemorrhagic pathology reappeared within 6 months but, apart from one case, repetition of the cycle using the same modalities as before, mastered and normalised the menstrual picture.