Reconstitution of huPBL-NSG Mice with Donor-Matched Dendritic Cells Enables Antigen-Specific T-cell Activation

Humanized mouse models provide a unique opportunity to study human immune cells in vivo, but traditional models have been limited to the evaluation of non-specific T-cell interactions due to the absence of antigen-presenting cells. In this study, immunodeficient NOD/SCID/IL2r-??^null (NSG) mice were engrafted with human peripheral blood lymphocytes alone or in combination with donor-matched monocyte-derived dendritic cells (DC) to determine whether antigen-specific T-cell activation could be reconstituted. Over a period of 3 weeks, transferred peripheral blood lymphocytes reconstituted the spleen and peripheral blood of recipient mice with predominantly human CD45-positive lymphocytes. Animals exhibited a relatively normal CD4/CD8 ratio (average 1.63:1) as well as reconstitution of CD3/CD56 (averaging 17.8%) and CD20 subsets (averaging 4.0%). Animals reconstituted with donor-matched CD11c+ DC also demonstrated a CD11c+ population within their spleen, representing 0.27% to 0.43% of the recovered human cells with concurrent expression of HLA-DR, CD40, and CD86. When immunized with adenovirus, either as free replication-incompetent vector (AdV) or as vector-transduced DC (DC/AdV), there was activation and expansion of AdV-specific T-cells, an increase in Th1 cytokines in serum, and skewing of T-cells toward an effector/memory phenotype. T-cells recovered from animals challenged with AdV in vivo proliferated and secreted a Th1-profile of cytokines in response to DC/AdV challenge in vitro. Our results suggest that engrafting NSG mice with a combination of lymphocytes and donor-matched DC can reconstitute antigen responsiveness and allow the in vivo assessment of human immune response to viruses, vaccines, and other immune challenges.     

Suicide and Homicide: Durkheim's and Henry & Short's Theories Tested on Data from the Baltic States

The three Baltic States - Estonia, Latvia and Lithuania - were studied in two sociopolitically divergent periods. The first was the stagnation period under the Soviet regime in 1970-1984. The second was the period of democratic reforms that started in 1985, with encouraging political changes, ensuing difficulties associated with the change-over to the market economy from 1989; and finally stabilization in 1994-1998. The results of our study supported the hypothesis that suicide and homicide are affected by sociopolitical and economic conditions. Durkheim's suicide theory affords an explanation of the stable high suicide rate during stagnation period and the subsequent S-shaped suicide trend (fall-rise fall) during the reform period. However, the parallel suicide and homicide trends in the Baltic States do not tally with the postulates of Durkheim and Henry & Short regarding the inverse correlation of suicide and homicide at various levels of external restraints and with the varying state of the economy.     

Suicide among older people in relation to their subjective and objective well-being in different European regions

The aim of this study was to establish how different types of welfare states shape the context of the everyday life of older people by influencing their subjective well-being, which in turn might manifest itself in suicide rates. Twenty-two European countries studied were divided into Continental, Nordic, Island, Southern, and post-socialist countries, which were subdivided into Baltic, Slavic, and Central-Eastern groups based on their socio-political and welfare organization. Suicide rates, subjective well-being data, and objective well-being data were used as parameters of different welfare states and obtained from the World Health Organization European Mortality Database, European Social Survey, and Eurostat Database. This study revealed that the suicide rates of older people were the highest in the Baltic countries, while in the Island group, the suicide rate was the lowest. The suicide rate ratios between the age groups 65+ and 0–64 were above 1 (from 1.2 to 2.5), except for the group of the Island countries with a suicide rate ratio of 0.8. Among subjective well-being indicators, relatively high levels of life satisfaction and happiness were revealed in Continental, Nordic, and Island countries. Objective well-being indicators like old age pension, expenditure on old age, and social protection benefits in GDP were the highest in the Continental countries. The expected inverse relationship between subjective well-being indicators and suicide rates among older people was found across the 22 countries. We conclude that welfare states shape the context and exert influence on subjective well-being, and thus may lead to variations in risk of suicide at the individual level.     

Work of breathing using different interfaces in spontaneous positive pressure ventilation: helmet,face-mask,and endotracheal tube

Purpose

Noninvasive positive pressure ventilation (NPPV) using a helmet is expected to cause inspiratory trigger delay due to the large collapsible and compliant chamber. We compared the work of breathing (WOB) of NPPV using a helmet or a full face-mask with that of invasive ventilation by tracheal intubation.

Methods

We used a lung model capable of simulating spontaneous breathing (LUNGOO; Air Water Inc., Japan). LUNGOO was set at compliance (C) = 50 mL/cmH₂O and resistance (R) = 5 cmH₂O/L/s for normal lung simulation, C = 20 mL/cmH₂O and R = 5 cmH₂O/L/s for restrictive lung, and C = 50 mL/cmH₂O and R = 20 cmH₂O/L/s for obstructive lung. Muscle pressure was fixed at 25 cmH₂O and respiratory rate at 20 bpm. Pressure support ventilation and continuous positive airway pressure were performed with each interface placed on a dummy head made of reinforced plastic that was connected to LUNGOO. We tested the inspiratory WOB difference between the interfaces with various combinations of ventilator settings (positive end-expiratory pressure 5 cmH₂O; pressure support 0, 5, and 10 cmH₂O).

Results

In the normal lung and restrictive lung models, WOB decreased more with the face-mask than the helmet, especially when accompanied by the level of pressure support. In the obstructive lung model, WOB with the helmet decreased compared with the other two interfaces. In the mixed lung model, there were no significant differences in WOB between the three interfaces.

Conclusion

NPPV using a helmet is more effective than the other interfaces for WOB in obstructive lung disease.

     

Neck muscles cross-sectional area in adolescents with and without headache – MRI study

Background: Cervical musculature may play an important role in the genesis of tension‐type headache. However, there are no reports on a possible association between the morphometrical features of the neck flexion and extension muscles and adolescence headache. Aim: To examine differences in neck flexion and extension muscles cross‐sectional area (CSA) in adolescents with and without headache. Methods: A population‐based sample of 17‐year‐old adolescents with migraine (N=19), tension‐type headache (N=24) and healthy controls without headache (N=22) was examined. CSA of the neck muscles was measured from axial T1‐weighted magnetic resonance images (MRI). Results: Boys with tension‐type headache showed significantly smaller CSA of right sternocleidomastoid muscle than boys with migraine and girls with tension‐type headache showed significantly smaller CSA of combined right sternocleidomastoid and scalenus muscles than girls with migraine. In addition, boys with migraine had significantly larger CSA of both right sternocleidomastoid and combined right sternocleidomastoid and scalenus muscles, and left semispinalis capitis muscle and combined left semispinalis and splenius muscles than boys without headache. In boys and girls no other significant differences were observed in the CSA of neck flexion or extension muscles. Conclusions: This preliminary work demonstrates that both girls and boys with tension‐type headache and migraine have differences in the size of neck flexion muscles, especially unilaterally. In boys, unilaterally increased size of neck flexion and extension muscles is associated with migraine. These findings, if confirmed in further studies, may have important diagnostic and therapeutic implications for rehabilitation of adolescents with headache.     

Real-time PCR analysis of enteric pathogens from fecal samples of irritable bowel syndrome subjects

Background

Growing amount of scientific evidence suggests that microbes are involved in the pathophysiology of irritable bowel syndrome (IBS). The predominant fecal microbiota composition of IBS subjects has been widely studied with DNA-based techniques but less research has been focused on the intestinal pathogens in this disorder. Here, we optimized a highly sensitive panel of 12 quantitative real-time PCR (qPCR) assays to shed light on the putative presence of intestinal pathogens in IBS sufferers. The panel was used to screen fecal samples from 96 IBS subjects and 23 healthy controls.

Results

Fifteen IBS samples (17%) tested positive for Staphylococcus aureus with a thermonuclease (nuc) gene-targeting qPCR assay, whereas none of the healthy controls were positive for S. aureus (p < 0.05). The S. aureus -positive IBS samples were confirmed by sequencing of the PCR amplicons. Clostridium perfringens was detected from IBS and control groups with a similar frequency (13% and 17%, respectively) with α-toxin (plc) gene -targeting qPCR assay while none of the samples tested positive for the Cl. perfringens enterotoxin-encoding gene (cpe).

Conclusions

The qPCR panel consisting of 12 assays for an extensive set of pathogenic microorganisms provides an efficient alternative to the conventional detection of gastrointestinal pathogens and could accelerate the initiation of targeted antibiotic therapy reducing the risk of post-infectious IBS (PI-IBS). S. aureus has not been previously reported to be associated with the onset of IBS. Although we discovered significant differences in the prevalence of S. aureus between the study groups, its importance in giving rise to IBS symptoms requires further studies.     

Longer Survival in Patients with Breast Cancer with Cyclin D1 Over-Expression after Tumor Recurrence: Longer,but Occupied with Disease

Purpose

The effect of cyclin D1 overexpression on breast cancer outcomes and prognosis is controversial, even though amplification of the cyclin D1 gene, CCND1, has been shown to be associated with early relapse and poor prognosis. In this study, we examined the relationship between cyclin D1 overexpression and disease-specific survival (DSS). We also analyzed survival in patients who experienced recurrence.

Methods

We retrospectively analyzed data from patients diagnosed with ductal carcinoma between April 2005 and December 2010. We examined clinicopathologic factors associated with cyclin D1 overexpression and analyzed the influence of cyclin D1 on recurrence-free survival and DSS.

Results

We identified 236 patients diagnosed with primary breast cancer who completed all phases of their primary treatment. Cyclin D1 overexpression was significantly associated with longer DSS (5-year DSS, 89.9% in patients without cyclin D1 overexpression vs. 98.9% in patients with cyclin D1 overexpression; p=0.008). Multivariate analysis also found that patients with cyclin D1 overexpressing tumors had significantly longer disease-specific survival than patients whose tumors did not overexpress cyclin D1, with a hazard ratio for disease-specific mortality of 7.97 (1.17-54.22, p=0.034). However, in the group of patients who experienced recurrence, cyclin D1 overexpression was not significantly associated with recurrence-free survival. Cyclin D1 overexpression was significantly associated with increased survival after disease recurrence, indicating that cyclin D1 overexpression might be indicative of more indolent disease progression after metastasis.

Conclusion

Cyclin D1 overexpression is associated with longer DSS, but not recurrence-free survival, in patients with breast cancer. Longer postrecurrence survival could explain the apparent inconsistency between DSS and recurrence-free survival. Patients with cyclin D1-overexpressing tumors survive longer, but with metastatic disease after recurrence. This information should spark the urgent development of tailored therapies to cure these patients.     

Adolescent subthreshold‐depression and anxiety: psychopathology,functional impairment and increased suicide risk

Background: Subthreshold‐depression and anxiety have been associated with significant impairments in adults. This study investigates the characteristics of adolescent subthreshold‐depression and anxiety with a focus on suicidality, using both categorical and dimensional diagnostic models. Methods: Data were drawn from the Saving and Empowering Young Lives in Europe (SEYLE) study, comprising 12,395 adolescents from 11 countries. Based on self‐report, including Beck Depression Inventory‐II (BDI‐II), Zung Self‐Rating Anxiety Scale (SAS), Strengths and Difficulties Questionnaire (SDQ) and Paykel Suicide Scale (PSS) were administered to students. Based on BDI‐II, adolescents were divided into three groups: nondepressed, subthreshold‐depressed and depressed; based on the SAS, they were divided into nonanxiety, subthreshold‐anxiety and anxiety groups. Analyses of Covariance were conducted on SDQ scores to explore psychopathology of the defined groups. Logistic regression analyses were conducted to explore the relationships between functional impairments, suicidality and subthreshold and full syndromes. Results: Thirty‐two percent of the adolescents were subthreshold‐anxious and 5.8% anxious, 29.2% subthreshold‐depressed and 10.5% depressed, with high comorbidity. Mean scores of SDQ of subthreshold‐depressed/anxious were significantly higher than the mean scores of the nondepressed/nonanxious groups and significantly lower than those of the depressed/anxious groups. Both subthreshold and threshold‐anxiety and depression were related to functional impairment and suicidality. Conclusions: Subthreshold‐depression and subthreshold‐anxiety are associated with an increased burden of disease and suicide risk. These results highlight the importance of early identification of adolescent subthreshold‐depression and anxiety to minimize suicide. Incorporating these subthreshold disorders into a diagnosis could provide a bridge between categorical and dimensional diagnostic models.