Quantitative Assessment of Optimal Bone Marrow Site for the Isolation of Porcine Mesenchymal Stem Cells.

Background. One of the most plentiful sources for MSCs is the bone marrow; however, it is unknown whether MSC yield differs among different bone marrow sites. In this study, we quantified cellular yield and evaluated resident MSC population from five bone marrow sites in the porcine model. In addition, we assessed the feasibility of a commercially available platelet concentrator (Magellan® MAR01™ Arteriocyte Medical Systems, Hopkinton, MA) as a bedside stem cell concentration device. Methods. Analyses of bone marrow aspirate (BMA) and concentrated bone marrow aspirate (cBMA) included bone marrow volume, platelet and nucleated cell yield, colony-forming unit fibroblast (CFU-F) number, flow cytometry, and assessment of differentiation potential. Results. Following processing, the concentration of platelets and nucleated cells significantly increased but was not significantly different between sites. The iliac crest had significantly less bone marrow volume; however, it yielded significantly more CFUs compared to the other bone marrow sites. Culture-expanded cells from all tested sites expressed high levels of MSC surface markers and demonstrated adipogenic and osteogenic differentiation potential. Conclusions. All anatomical bone marrow sites contained MSCs, but the iliac crest was the most abundant source of MSCs. Additionally, the Magellan can function effectively as a bedside stem cell concentrator.

Transplantation and perfusion of microvascular fragments in a rodent model of volumetric muscle loss injury.

Few clinical options are available for the treatment of volumetric muscle loss (VML). An important consideration that needs to be addressed for the development of treatments for these injuries is the establishment of a vascular supply sufficient to support skeletal muscle regeneration. The objective of the current study was to evaluate the potential for microvascular fragments (MVFs) harvested from adipose tissue to support tissue perfusion for VML. Tibialis anterior muscle defects in rats were replaced with constructs that were created on the day of surgery containing either (1) collagen only (COL), (2) freshly isolated microvascular fragments in collagen (MVF), or (3) adipose tissue derived stem cells (ASCs) in collagen. Muscles were harvested 7 and 14 days after surgery. Defects treated with MVFs had a vessel density higher than the other groups at both 7 and 14 days, and those treated with ASCs had a higher vessel density than COL by day 14 (p < 0.05). Perfused vessels were observed in both the ASC and MVF treated defects at day 14, as well as at day 7 in the MVF. This study supports the use of MVFs as a platform to improve tissue perfusion to treat large VML defects. The use of freshly isolated MVFs on the day of surgery supports their clinical use and application.

Mutational analysis of AXIN2, MSX1, and PAX9 in two Mexican oligodontia families.

The genes for axin inhibition protein 2 (AXIN2), msh homeobox 1 (MSX1), and paired box gene 9 (PAX9) are involved in tooth root formation and tooth development. Mutations of the AXIN2, MSX1, and PAX9 genes are associated with non-syndromic oligodontia. In this study, we investigated phenotype and AXIN2, MSX1, and PAX9 gene variations in two Mexican families with non-syndromic oligodontia. Individuals from two families underwent clinical examinations, including an intra-oral examination and panoramic radiograph. Retrospective data were reviewed, and peripheral blood samples were collected. The exons and exon-intronic boundaries of the AXIN2, MSX1, and PAX9 genes were sequenced and analyzed. Protein and messenger RNA structures were predicted using bioinformative software programs. Clinical and oral examinations revealed isolated non-syndromic oligodontia in the two Mexican families. The average number of missing teeth was 12. The sequence analysis of exons and exon-intronic regions of AXIN2, MSX1, and PAX9 revealed 11 single-nucleotide polymorphisms (SNPs), including seven in AXIN2, two in MSX1, and three in PAX9. One novel SNP of MSX1, c.476T>G (Leu159Arg), was found in all of the studied patients in the families. MSX1 Leu159Arg and PAX9 Ala240Pro change protein and messenger RNA structures. Our findings suggested that a combined reduction of MSX1 and PAX9 gene dosages increased the risk for oligodontia in the Mexican families, as in vivo investigation has indicated that interaction between Msx1 and Pax9 is required for tooth development.

Autologous minced muscle grafts: a tissue engineering therapy for the volumetric loss of skeletal muscle.

Volumetric muscle loss (VML) results in a large void deficient in the requisite materials for regeneration for which there is no definitive clinical standard of care. Autologous minced muscle grafts (MG), which contain the essential components for muscle regeneration, may embody an ideal tissue engineering therapy for VML. The purpose of this study was to determine if orthotopic transplantation of MG acutely after VML in the tibialis anterior muscle of male Lewis rats promotes functional tissue regeneration. Herein we report that over the first 16 wk postinjury, MG transplantation 1) promotes remarkable regeneration of innervated muscle fibers within the defect area (i.e., de novo muscle fiber regeneration); 2) reduced evidence of chronic injury in the remaining muscle mass compared with nonrepaired muscles following VML (i.e., transplantation attenuated chronically upregulated transforming growth factor-ß1 gene expression and the presence of centrally located nuclei in 30% of fibers observed in nonrepaired muscles); and 3) significantly improves net torque production (i.e., ∼55% of the functional deficit in nonrepaired muscles was restored). Additionally, voluntary wheel running was shown to reduce the heightened accumulation of extracellular matrix deposition observed within the regenerated tissue of MG-repaired sedentary rats 8 wk postinjury (collagen 1% area: sedentary vs. runner, ∼41 vs. 30%), which may have been the result of an augmented inflammatory response [i.e., M1 (CCR7) and M2 (CD163) macrophage expression was significantly greater in runner than sedentary MG-repaired muscles 2 wk postinjury]. These findings support further exploration of autologous minced MGs for the treatment of VML.

Phenotype characterization and sequence analysis of BMP2 and BMP4 variants in two Mexican families with oligodontia.

Both BMP2 and BMP4 are involved in tooth development. We examined phenotypes and BMP2 and BMP4 gene variations in two Mexican oligodontia families. Physical and oral examinations and panoramic radiographs were performed on affected and unaffected members in these two families. The affected members lacked six or more teeth. DNA sequencing was performed to detect BMP2 and BMP4 gene variations. Three single nucleotide polymorphisms (SNPs) in BMP2 and BMP4 genes were identified in the two families, including one synonymous and two missense SNPs: BMP2 c261A>G, pS87S, BMP2 c570A>T, pR190S, and BMP4 c455T>C, pV152A. Among the six affected patients, 67% carried "GG" or "AG" genotype in BMP2 c261A>G and four were "TT" or "AT" genotype in BMP2 c570A>T (pR190S). Polymorphism of BMP4 c455T>C resulted in amino acid changes of Val/Ala (pV152A). BMP2 c261A>G and BMP4 c455T>C affect mRNA stability. This was the first time that BMP2 and BMP4 SNPs were observed in Mexican oligodontia families.

Predictors of psychological adjustment in school-age children infected with HIV.

OBJECTIVE: To assess for significant differences in psychological functioning between HIV-infected children and a demographically matched healthy control group and to examine the utility of applying a stress and coping model to children with HIV disease. METHODS: Participants included HIV-infected children (ages 6-16) and their caregivers (n = 36) and a control group of healthy children and their caregivers (n = 32). During routine clinic visits, children completed measures of psychological adjustment, health locus of control, and coping style, and caregivers completed measures of their own and their child's psychological adjustment. RESULTS: Caregiver-reported and child self-reported psychological adjustment scores did not significantly differ between the HIV and control groups, with the exception of significantly more internalizing behavior problems reported in the control group. Hierarchical multiple regression analyses revealed that the stress and coping model accounted for 36% of the variance in HIV-infected children's self-reported psychological adjustment. In addition, child age and coping style were significant predictors of child self-reported psychological adjustment, but not of caregiver-reported child adjustment. CONCLUSIONS: Approximately 25% of children with HIV disease exhibited clinically significant emotional or behavioral problems; however, even higher rates of psychological adjustment problems were found in healthy children. Children with HIV disease who have not been told their diagnosis and children who endorse more emotion-focused coping strategies tend to exhibit more psychological adjustment problems.

Psychological adjustment in caregivers of school-age children infected with HIV: stress, coping, and family factors.

OBJECTIVE: To assess for significant differences in psychological functioning between caregivers of HIV-infected children and caregivers of healthy children, and to examine the utility of applying a stress and coping model to caregivers of children with HIV disease. METHODS: Participants included caregivers of HIV-infected children (n = 36) and caregivers of a demographically matched control group of healthy children (n = 32). During their child's pediatric clinic visits, caregivers completed measures of psychological adjustment, stress, coping style, and family resources and support. They also completed a measure of their child's psychological adjustment. RESULTS: Caregiver psychological distress scores did not significantly differ between the HIV and control groups, and clinically significant rates of psychological distress were reported by more than a third of caregivers in both groups. Hierarchical multiple regression analyses revealed that independent of their child's illness status, stress and coping style were significant predictors of caregiver's psychological adjustment. In addition, caregiver psychological distress was a significant predictor of children's maladjustment. CONCLUSIONS: High rates of psychological distress were observed in caregivers of children with HIV disease; however, similarly high rates of psychological adjustment problems were found in caregivers of healthy children. Caregivers who reported high levels of daily stress and emotion-focused coping styles tended to report more psychological distress. Further, caregivers who reported more psychological distress also reported more internalizing and more externalizing behavior problems in their children, regardless of the child's illness status. These findings reflect the impact of poverty and environmental stress on caregivers' adjustment.

Higher than normal plasma interleukin-6 concentrations in cancer patients with depression: preliminary findings.

OBJECTIVE: This study investigated whether cancer patients with and without major depression exhibit immune system abnormalities similar to those reported in medically healthy, depressed subjects without cancer. METHOD: The study subjects consisted of patients diagnosed with pancreatic, esophageal, or breast cancer. Other groups consisted of subjects with major depression (without cancer) and healthy comparison subjects. Subjects' diagnoses were made with the Structured Clinical Interview for DSM-III-R. Severity of depression was measured with the Hamilton Depression Rating Scale. Plasma concentrations of interleukin-6 (IL-6) and postdexamethasone cortisol were measured. RESULTS: Cancer patients with depression had markedly higher plasma concentrations of IL-6 than healthy comparison subjects and cancer patients without depression. Although significant correlations were found between Hamilton depression scale scores and plasma concentrations of postdexamethasone cortisol, no significant correlations were found between plasma IL-6 and postdexamethasone cortisol concentrations. CONCLUSIONS: Higher than normal plasma IL-6 concentrations were associated with a diagnosis of major depression in cancer patients. IL-6 may contribute to sickness behavior that has overlapping symptoms with major depression.

Mood and anxiety disorders.

This chapter describes two of the most frequent psychiatric conditions occurring in people with HIV and AIDS, namely, anxiety disorders and depression, and describes the commonly occurring features, clinical correlates, and treatment considerations for these disorders.

Resilience factors associated with adaptation to HIV disease.

This study examines the hardiness dimensions of commitment, challenge, and control as resilience factors in adaptation among persons with symptomatic HIV disease and AIDS. Two hundred participants completed self-report questionnaires measuring hardiness, psychological distress, quality of life, and core personal beliefs. A series of standard multiple regression analyses revealed that high hardiness was significantly related to 1) lower psychological distress levels; 2) higher perceived quality of life in physical health, mental health, and overall functioning domains; 3) more positive personal beliefs regarding the benevolence of the world and people, self-worth, and randomness of life events; and 4) lowered belief in controllability of life events. Commitment was the hardiness factor that most frequently made a unique contribution to predicting adaptation in the regression models. Implications of these findings for understanding HIV-related adaptation and for clinical mental health intervention are considered. Future directions in HIV-related adaptation research are suggested.

Double-blind comparison of fluoxetine and desipramine in the treatment of depressed women with advanced HIV disease: a pilot study.

A double-blind, placebo-controlled study was conducted to assess the relative efficacy and tolerability of fluoxetine and desipramine in depressed, human immunodeficiency virus (HIV)-positive women. Although difficulty in the recruitment and retention of participants led to insufficient power to detect differences between treatment groups, results indicated that participants experienced improvement in their depression. However, for most women, significant depressive symptoms remained after 6 weeks of treatment. In addition, although most participants reported at least one adverse event after treatment began, most of the side effects, regardless of treatment condition, were mild to moderate in severity. Important barriers to study participation and completion are discussed, as well as suggestions for increasing the involvement of depressed, HIV-positive women in future treatment studies.

A National, Randomized Survey of HIV/AIDS Knowledge and Attitudes Among Psychiatrists in Training.

Given the broad spectrum of medical and psychiatric complications associated with HIV (human immunodeficiency virus) disease, HIV-related education is an important curriculum topic for all physicians. This study examined the HIV-related knowledge and attitudes among psychiatrists in training to evaluate their training needs. A survey instrument was developed containing 15 knowledge items, 15 attitude items, demographic items, and questions concerning how HIV/AIDS (acquired immunodeficiency syndrome) affected residency choice and self-perception of training needs. The questionnaire was mailed to 2,252 psychiatrists in training randomly selected and stratified by American Psychiatric Association district and postgraduate training year. The sample consisted of 825 respondents representing 37% of the initial pool. Knowledge was most lacking in areas dealing with HIV-related neuropsychiatric complications and issues concerning HIV/AIDS and special populations. In general, the respondents expressed positive attitudes about HIV-related issues; however, three items correctly endorsed by fewer than 75% of the sample dealt specifically with the management of neuropsychiatric complications, working with dying patients, and the range of normal sexuality. Targeted HIV-related education is needed for psychiatrists in training and particularly should encompass neuropsychiatric syndromes and issues of special populations.

Severe mental illness and HIV-related medical and neuropsychiatric sequelae.

Medical and neuropsychiatric sequelae of HIV infection present a spectrum of diagnostic and treatment challenges to mental health clinicians. Both HIV and the many opportunistic infections that manifest in patients due to their immunocompromised state also can affect the central nervous system (CNS). Thus, mental health clinicians need to be familiar with the diagnosis and management of HIV-related medical and psychiatric complications. This article provides an overview of the CNS-related manifestations resulting from HIV disease, including HIV-related dementia, psychotic disorders, delirium, CNS opportunistic infections and tumors, systemic abnormalities, psychoactive substances, and the adverse effects of certain medical treatments. Treatment strategies for individuals with HIV disease and comorbid severe mental illness are outlined and recommendations for future research are offered.

An assessment of rates of psychiatric morbidity and functioning in HIV disease.

This study examined demography, rates of psychopathology, and functional impairment in HIV-seropositive women and men in a large, urban, public outpatient infectious disease clinic. Fifty-three percent of the women and 70% of the men met Structural Clinical Interview for DSM-III-R criteria for psychiatric disorders. Current mood disorders were the most frequent diagnoses, followed by psychoactive substance abuse/dependence disorders and psychotic disorders. Seventy-six percent of the women and 90% of the men had previous psychiatric histories, including 59% of the women and 55% of the men who had psychiatric histories prior to their knowledge of HIV seroconversion. Depressed subjects reported significant impairment in physical, social, and role functioning. Similarly, impairment in physical functioning was highly correlated with self-reported anxiety symptoms. These data suggest considerable past and current psychiatric comorbidity in HIV-infected individuals seeking medical care, and draw attention to the need for recognition and aggressive psychiatric treatment, particularly for those depressed and anxious patients with impaired functioning.

A survey of knowledge and attitudes about HIV and AIDS among medical students.

Researchers have found that a significant number of medical students harbor attitudes that have a negative impact on their willingness to care for persons who are HIV positive or have AIDS. To assess current HIV and AIDS knowledge and attitudes, the authors administered a 25-item survey tailored for medical professionals to 63 preclinical medical students. Respondents' mean score on the knowledge scale was 6.25 (SD 1.63) out of a possible score of 10. Factor analysis suggested three major groupings of medical students with regard to attitudes. The largest group had generally positive attitudes about patients with HIV and AIDS; two subgroups, however, would tend to refer such patients to another physician. One subgroup reported feeling more uncomfortable with homosexual behavior and with HIV-seropositive patients than they did with patients with other infectious diseases. This group also expressed discomfort with physically touching HIV-seropositive clients. The other group expressed discomfort with taking a patient's sexual history. Effective educational interventions must take these findings into account.

Depression in patients with cancer. Diagnosis, biology, and treatment.

This article reviews the challenge of diagnosing depression in patients with cancer. Major depression and depressive symptoms, although commonly encountered in medical populations, are frequently underdiagnosed and undertreated. This is especially true for patients with cancer in whom the diagnosis of major depression is clouded by neurovegetative symptoms that may be secondary to either cancer or depression. Well-established biological markers for major depression are proposed as diagnostic adjuncts in patients with cancer. Studies using biological markers in depressed patients with and without cancer are reviewed, and the implications of diminished immune function in depressed patients with cancer are discussed. The limited database on treatment of depression in patients with cancer also is reviewed. Treatment of depression in these patients improves their dysphoria and other signs and symptoms of depression, improves quality of life, and may improve immune function and survival time. Guidelines for future research are proposed.