Functionally distinct human marrow stromal cell lines immortalized by transduction with the human papilloma virus E6/E7 genes

A replication-defective recombinant retrovirus containing the human papilloma virus E6/E7 genes (LXSN-16 E6E7) was used to immortalize stromal cells from human marrow. The E6/E7 gene products interfere with the function of tumor-suppressor proteins p53 and Rb, respectively, thereby preventing cell cycle arrest without causing significant transformation. Twenty-seven immortalized clones designated HS-1 to HS- 27 were isolated, four of which are characterized in this report. Two cell lines, HS-5 and HS-21, appear to be fibroblastoid and secrete significant levels of granulocyte colony-stimulating factor (G-CSF), granulocyte-macrophage-CSF (GM-CSF), macrophage-CSF (M-CSF), Kit ligand (KL), macrophage-inhibitory protein-1 alpha, interleukin-6 (IL-6), IL- 8, and IL-11. However, only HS-5 supports proliferation of hematopoietic progenitor cells when cocultured in serum-deprived media with no exogenous factors. Conditioned media (CM) from HS-5 promotes growth of myeloid colonies to significantly greater extent than a cocktail of recombinant factors containing 10 ng/mL of IL-1, IL-3, IL- 6, G-CSF, GM-CSF, and KL and 3 U of erythropoietin (Epo). Two additional clones, HS-23 and HS-27, resemble "blanket" cells, with an epithelioid morphology, and are much larger, broader, and flatter when compared with HS-5 and HS-21. These lines secrete low levels of growth factors and do not support proliferation of isolated progenitor cells in cocultures. CM from HS-23 and HS-27 also fail to support growth of myeloid colonies. Both HS-23 and HS-27 express relatively high levels of VCAM-1, yet HS-27 is the only line that supports the formation of "cobblestone" areas by isolated CD34+38lo cells. We hypothesize that HS- 5, HS-21, HS-23, and HS-27 represent functionally distinct components of the marrow microenvironment.     

Nonopsonic uptake of Mycobacterium avium complex by human monocytes and alveolar macrophages.

The uptake of Mycobacterium avium complex (MAC) microorganisms by human peripheral blood monocytes (PBMs) and alveolar macrophages (AMs) is not well understood. We have previously shown, under opsonic conditions, that humoral factors are important in mediating the uptake of MAC by PBMs. However, the receptor-ligand interactions occurring under nonopsonic conditions remain unclear. We compared the uptake of untreated human PBMs and AMs in a serum-free medium with phagocytes treated to remove surface receptors. Removal of complement receptors CR1 and CR3, the Fc receptor (FcR), and the transferrin receptor (TfR) resulted in significantly lower levels of MAC uptake in serum-free medium by both PBMs and AMs. The addition of barley beta-glucan or mannan from Saccharomyces cerevisiae inhibited MAC uptake by untreated phagocytes in a dose-dependent manner. MAC uptake by PBMs or AMs was never completely abrogated by combining treatments (removal of CR1, CR3, FcR, and TfR and adding mannan or beta-glucan), indicating still-unknown mechanisms of uptake under nonopsonic conditions. We conclude that CR1, CR3, FcR, TfR, the mannose receptor, and possibly a separate beta-glucan-inhibitable receptor all may be involved in nonopsonic uptake of MAC by both PBMs and AMs.     

The tus gene of Escherichia coli: autoregulation, analysis of flanking sequences and identification of a complementary system in Salmonella typhimurium

The tus gene of Escherichia coli encodes a DNA-binding protein that, when bound to terminator sites, blocks replication forks. One of these sites, TerB, is immediately upstream from tus, and we have determined that the 5' end of tus mRNA is in the TerB site, that tus is autoregulated and that pTus is a very low efficiency promoter. Analysis of the DNA upstream from tus and TerB indicates a set of sensor/regulator genes which are comparable to envZ/ompR. Although tus mutants exhibit no growth phenotype in laboratory conditions, Salmonella typhimurium and E. coli have nevertheless maintained similar termination systems. Sequence homology can be demonstrated by Southern hybridizations, and the systems also exhibit functional complementation: the Tus protein of S. typhimurium blocks DNA replication at the TerA site of E. coli.     

Altered in vitro Handling of Mycobacterium Avium Complex by Monocytes and Serum from HIV(+) Patients

In patients with acquired immunodeficiency syndrome (AIDS), mycobacterial diseases are leading opportunistic infections. The reasons for the peculiar propensity for disseminated infection with Mycobacterium avium complex (MAC) remain unclear. We have previously examined, in detail, the ability of monocytes from healthy donors to take up and kill MAC under both nonopsonic and opsonic conditions. We have now evaluated the in vitro ability of peripheral blood monocytes from HIV(+) patients to take up and kill MAC organisms, and have discovered a reduced ability under both nonopsonic and opsonic conditions. This reduction is due to: 1) apparent defect (s) in the phagocytes themselves, and 2) substance (s) in the HIV(+) serum which actively suppresses phagocyte activity.     

Photoperiod is associated with hippocampal volume in a large community sample

Although animal research has demonstrated seasonal changes in hippocampal volume, reflecting seasonal neuroplasticity, seasonal differences in human hippocampal volume have yet to be documented. Hippocampal volume has also been linked to depressed mood, a seasonally varying phenotype. Therefore, we hypothesized that seasonal differences in day‐length (i.e., photoperiod) would predict differences in hippocampal volume, and that this association would be linked to low mood. Healthy participants aged 30–54 (M = 43; SD = 7.32) from the University of Pittsburgh Adult Health and Behavior II project (n = 404; 53% female) were scanned in a 3T MRI scanner. Hippocampal volumes were determined using an automated segmentation algorithm using FreeSurfer. A mediation model tested whether hippocampal volume mediated the relationship between photoperiod and mood. Secondary analyses included seasonally fluctuating variables (i.e., sleep and physical activity) which have been shown to influence hippocampal volume. Shorter photoperiods were significantly associated with higher BDI scores (R² = 0.01, β = −0.12, P = 0.02) and smaller hippocampal volumes (R² = 0.40, β = 0.08, P = 0.04). However, due to the lack of an association between hippocampal volume and Beck Depression Inventory scores in the current sample, the mediation hypothesis was not supported. This study is the first to demonstrate an association between season and hippocampal volume. These data offer preliminary evidence that human hippocampal plasticity could be associated with photoperiod and indicates a need for longitudinal studies. © 2014 Wiley Periodicals, Inc.     

Cognitive-behavioral therapy, light therapy, and their combination in treating seasonal affective disorder

Background: The need to develop supplementary or alternative treatments for seasonal affective disorder (SAD) is underscored by the significant minority (47%) of SAD patients that is refractory to light therapy, the persistence of residual symptoms despite light treatment, and poor long-term compliance with light use. Because preliminary studies suggest that cognitive and behavioral factors are involved in SAD, cognitive-behavioral therapy (CBT) warrants investigation as a possible treatment option. Methods: We piloted a 6-week randomized clinical trial to compare a standard light therapy protocol; a novel, SAD-tailored, group CBT intervention; and their combination in ameliorating and remitting a current SAD episode and as prophylaxis against episode recurrence. Depressive symptom severity and remission rates were assessed at post-treatment and at a 1-year follow-up visit to examine long-term treatment durability. Results: CBT, light therapy, and their combination all demonstrated significant reductions in depressive symptoms on two different outcome measures. Remission rates varied by measure, but did not reach statistical significance. During the subsequent winter, CBT, particularly in combination with light therapy, appeared to improve long-term outcome regarding symptom severity, remission rates, and relapse rates. No CBT-treated participant, with or without light, experienced a full SAD relapse compared to over 60% of those treated with light alone. Limitations: These results should be viewed as preliminary and are limited by the small sample size (n=23) and lack of a control group. Conclusions: The nearly half of SAD patients who do not remit with light alone may benefit from CBT as an adjunct or alternative treatment, especially as a prophylaxis against episode recurrence.     

The role of beliefs and attitudes about sleep in seasonal and nonseasonal mood disorder,and nondepressed controls

Background

Unhelpful sleep-related cognitions play an important role in insomnia and major depressive disorder, but their role in seasonal affective disorder has not yet been explored. Therefore, the purpose of this study was to determine if individuals with seasonal affective disorder (SAD) have sleep-related cognitions similar to those with primary insomnia, and those with insomnia related to comorbid nonseasonal depression.

Methods

Participants (n=147) completed the Dysfunctional Beliefs and Attitudes about Sleep 16-item scale (DBAS-16) and the Structured Interview Guide for the Hamilton Depression Rating Scale, Seasonal Affective Disorder Version (SIGH-SAD), which assesses self reported sleep problems including early, middle, or late insomnia, and hypersomnia in the previous week. All participants were assessed in winter, and during an episode for those with a depressive disorder.

Results

Individuals with SAD were more likely to report hypersomnia on the SIGH-SAD, as well as a combined presentation of hypersomnia and insomnia on the Pittsburgh Sleep Quality Index (PSQI). The SAD group reported DBAS-16 scores in the range associated with clinical sleep disturbance, and DBAS-16 scores were most strongly associated with reports of early insomnia, suggesting circadian misalignment.

Limitations

Limitations include the self-report nature of the SIGH-SAD instrument on which insomnia and hypersomnia reports were based.

Conclusions

Future work could employ sleep- or chronobiological-focused interventions to improve clinical response in SAD.     

Transketolase abnormality in cultured fibroblasts from familial chronic alcoholic men and their male offspring.

We have investigated a thiamine-dependent enzyme, transketolase, in cultured fibroblasts from 41 human subjects, including patients with alcoholism-associated Wernicke-Korsakoff syndrome (n = 3), familial chronic alcoholic males (n = 7), their sons (n = 7), nonalcoholic men (n = 7), their male offspring (n = 7), and three generations of an Amish family (n = 10) without any history of alcoholism. This study was undertaken to delineate whether transketolase abnormality (i.e., high Michaelis Menton constant (Km) for thiamine pyrophosphate), previously reported in patients with Wernicke-Korsakoff syndrome is prevalent among familial chronic alcoholic men and their sons without prior history of alcohol abuse but who are at high risk for alcoholism. Our data suggest that an inborn error (i.e., high Km of transketolase for thiamine pyrophosphate) predisposing to thiamine deficiency diseases similar to those reported in Wernicke-Korsakoff syndrome may occur in the general population. However, for some as yet unexplained reason(s) this variant seems to occur more frequently among familial chronic alcoholic men and their male offspring without any history of alcoholism. The inheritance pattern of this enzyme variant as revealed from an Amish pedigree study may be autosomal recessive as previously suggested.     

Cognitive vulnerability in moderate, mild, and low seasonality

This study examined the association between cognitive vulnerability factors and seasonality. Students (N = 88), classified based on the Seasonal Pattern Assessment Questionnaire as experiencing moderate (n = 26) or mild (n = 32) seasonality, and nondepressed, low-seasonality controls (n = 30) completed explicit (i.e., dysfunctional attitudes, automatic negative thoughts, seasonal attitudes, and rumination) and implicit (i.e., implicit associations test) measures of cognitive vulnerability at one winter and one nonwinter assessment. Relative to low- and mild-seasonality participants, moderate-seasonality participants endorsed more automatic thoughts and rumination in winter and more dysfunctional attitudes across both seasons. Moderate- and mild-seasonality participants endorsed more maladaptive seasonal attitudes than did low-seasonality participants. All groups demonstrated increased dysfunctional attitudes, automatic thoughts, and rumination and stronger implicit associations about light and dark during the winter. The findings support a possible cognitive mechanism of winter depression onset and/or maintenance unique to individuals with moderate, as opposed to mild, seasonality.     

Using Standardized Nursing Languages in End‐of‐Life Care Plans

PURPOSE: This case study demonstrates the challenges to achieve dignified life closure and a comfortable death for a middle‐aged woman with terminal cancer and her family. DATA SOURCES: Data were obtained from a patient known through the author's clinical experiences, personal family experiences, and published sources. DATA SYNTHESIS: The appropriate nursing diagnosis, patient outcomes, and nursing interventions were identified through the use of NANDA‐International, the Nursing Outcomes Classification, and the Nursing Interventions Classification. CONCLUSIONS: This case study illustrates the appropriate nursing diagnosis, interventions, and outcomes pertinent to an individual with emotional distress at the end of life. IMPLICATIONS FOR NURSING PRACTICE: Employing the NANDA‐International standardized nursing diagnosis, the Nursing Interventions Classification and the Nursing Outcomes Classification provided the needed constructs for considering and improving a dying patient's care in a primary and home setting.