Biologically derived epicardial patch induces macrophage mediated pathophysiologic repair in chronically infarcted swine hearts.

There are nearly 65 million people with chronic heart failure (CHF) globally, with no treatment directed at the pathologic cause of the disease, the loss of functioning cardiomyocytes. We have an allogeneic cardiac patch comprised of cardiomyocytes and human fibroblasts on a bioresorbable matrix. This patch increases blood flow to the damaged heart and improves left ventricular (LV) function in an immune competent rat model of ischemic CHF. After 6 months of treatment in an immune competent Yucatan mini swine ischemic CHF model, this patch restores LV contractility without constrictive physiology, partially reversing maladaptive LV and right ventricular remodeling, increases exercise tolerance, without inducing any cardiac arrhythmias or a change in myocardial oxygen consumption. Digital spatial profiling in mice with patch placement 3 weeks after a myocardial infarction shows that the patch induces a CD45pos immune cell response that results in an infiltration of dendritic cells and macrophages with high expression of macrophages polarization to the anti-inflammatory reparative M2 phenotype. Leveraging the host native immune system allows for the potential use of immunomodulatory therapies for treatment of chronic inflammatory diseases not limited to ischemic CHF.

Injected matrix stimulates myogenesis and regeneration of mouse skeletal muscle after ischaemic injury.

Biomaterial-guided regeneration represents a novel approach for the treatment of myopathies. Revascularisation and the intramuscular extracellular matrix are important factors in stimulating myogenesis and regenerating muscle damaged by ischaemia. In this study, we used an injectable collagen matrix, enhanced with sialyl LewisX (sLeX), to guide skeletal muscle differentiation and regeneration. The elastic properties of collagen and sLeX-collagen matrices were similar to those of skeletal muscle, and culture of pluripotent mESCs on the matrices promoted their differentiation into myocyte-like cells expressing Pax3, MHC3, myogenin and Myf5. The regenerative properties of matrices were evaluated in ischaemic mouse hind-limbs. Treatment with the sLeX-matrix augmented the production of myogenic-mediated factors insulin-like growth factor (IGF)-1, and IGF binding protein-2 and -5 after 3 days. This was followed by muscle regeneration, including a greater number of regenerating myofibres and increased transcription of Six1, M-cadherin, myogenin and Myf5 after 10 days. Simultaneously, the sLeX-matrix promoted increased mobilisation and engraftment of bone marrow-derived progenitor cells, the development of larger arterioles and the restoration of tissue perfusion. Both matrix treatments tended to reduce maximal forces of ischaemic solei muscles, but sLeX-matrix lessened this loss of force and also prevented muscle fatigue. Only sLeX-matrix treatment improved mobility of mice on a treadmill. Together, these results suggest a novel approach for regenerative myogenesis, whereby treatment only with a matrix, which possesses an inherent ability to guide myogenic differentiation of pluripotent stem cells, can enhance the endogenous vascular and myogenic regeneration of skeletal muscle, thus holding promise for future clinical use.

Application of Different Commercial Tourniquets by Laypersons: Would Public-access Tourniquets Work Without Training?

BACKGROUND: The White House "Stop the Bleed" campaign has renewed interest in public-access bleeding kits and the use of tourniquets by the lay public. The objective of this study was to determine which type of tourniquet could be applied most effectively by the lay public using only manufacturer instructions included with each tourniquet. METHODS: This prospective study randomized participants to one of four different tourniquets (SOFTT-W, CAT, RMT, SWAT-T). Participants were all over 18 years of age. Individuals with prior military, EMS, or patient-care medical experience were excluded. Using only the manufacturer's packaging instructions, participants were asked to apply a tourniquet on a simulated bleeding arm. A trained observer noted if tourniquet application by the participant was effective, partially effective, or ineffective based on reduction or cessation of simulated blood flow. Participant's application of the tourniquet was also timed (in seconds) by the observer. The primary outcome of our study was the effectiveness of application for each of the four tourniquets. Secondary outcome was time to effective application. RESULTS: A total of 176 participants were enrolled. For untrained laypersons the RMT had the highest effective application rate of 64.4% and was also the most rapidly applied at 100.9 ± 8.8 seconds (95% confidence interval [CI] = 83.1 to 118.6). The SWAT-T had the highest ineffective application rate (55.5%) than any other tourniquet type (p = 0.002). There was no effect of age or education on time to application for any tourniquet type. Effective applications were performed significantly faster than partially effective or ineffective applications (93.4 ± 5.8 [95% CI = 81.7 to 104.9] vs. 136.7 ± 8.7 [95% CI = 118.8 to 154.7] vs. 151.9 ± 8.3 [95% CI = 135.2 to 168.6]; p ≤ 0.001). There was no difference in time between partial and ineffective applications (p = 0.261). CONCLUSIONS: Our study suggests that laypersons could benefit from prior training to effectively apply tourniquets in emergency situations. Of the tourniquets studied, the RMT was the most effectively and most rapidly applied.

Alternatively spliced products CC3 and TC3 have opposing effects on apoptosis.

The human gene CC3 is a metastasis suppressor for small cell lung carcinoma (SCLC) in vivo. The ability of CC3 to impair the apoptotic resistance of tumor cells is likely to contribute to metastasis suppression. We describe here an alternatively spliced RNA of CC3, designated TC3, that encodes an unstable protein with antiapoptotic activity. TC3 and CC3 proteins share amino-terminal sequences, but TC3 has a unique short hydrophobic carboxyl terminus. Overexpression of CC3 results in massive death of rodent fibroblasts, but TC3 protects cells from CC3-induced death and from other death stimuli such as treatment with tumor necrosis factor or overexpression of Bax protein. The death-inducing activity of CC3 resides within its amino-terminal domain, which is conserved in TC3. The carboxyl terminus of TC3 is responsible for the antiapoptotic function of TC3; mutations in this domain abolish the ability of TC3 to protect cells from apoptosis. TC3 protein is short-lived due to its rapid degradation by proteasome, and it forms complexes with a regulatory subunit of proteasome known as s5alpha. The signal for the rapid degradation of TC3 resides within its carboxyl terminus, which is capable of conferring instability on a heterologous protein. The proapoptotic activity of CC3 in SCLC cells is induced by a wide variety of signals and involves disruption of the mitochondrial membrane potential (Deltapsim). The CC3 protein has sequence similarity to bacterial short-chain dehydrogenases/reductases and might represent a phylogenetically old effector of cell death similar to the recently identified apoptosis-inducing factor. CC3 and TC3 have opposing functions in apoptosis and represent a novel dual regulator of cell death.

Coordinate transcriptional and translational regulation of ferritin in response to oxidative stress.

The global increase in transcription of cytoprotective genes induced in response to oxidative challenge has been termed the antioxidant response. Ferritin serves as the major iron-binding protein in nonhematopoietic tissues, limiting the catalytic availability of iron for participation in oxygen radical generation. Here we demonstrate that ferritin is a participant in the antioxidant response through a genetically defined electrophile response element (EpRE). The EpRE of ferritin H identified in this report exhibits sequence similarity to EpRE motifs found in antioxidant response genes such as those encoding NAD(P)H:quinone reductase, glutathione S-transferase, and heme oxygenase. However, the EpRE of ferritin H is unusual in structure, comprising two bidirectional motifs arranged in opposing directions on complementary DNA strands. In addition to EpRE-mediated transcriptional activation, we demonstrate that ferritin is subject to time-dependent translational control through regulation of iron-regulatory proteins (IRP). Although IRP-1 is initially activated to its RNA binding (ferritin-repressing) state by oxidants, it rapidly returns to its basal state. This permits the translation of newly synthesized ferritin transcripts and ultimately leads to increased levels of ferritin protein synthesis following oxidant exposure. Taken together, these results clarify the complex transcriptional and translational regulatory mechanisms that contribute to ferritin regulation in response to prooxidant stress and establish a role for ferritin in the antioxidant response.

The partial nontandem duplication of the MLL (ALL1) gene is a novel rearrangement that generates three distinct fusion transcripts in B-cell acute lymphoblastic leukemia.

A partial nontandem duplication (PNTD) of mixed lineage leukemia (MLL) gene is described in B-cell acute lymphoid leukemia without structural cytogenetic abnormalities at 11q23 and 9p22. A duplicated portion of MLL is interrupted by the insertion of a region of 9p22 that includes the 3'-end of the AF9 gene. The PNTD encodes: (a) a PNTD transcript; (b) a partial tandem duplication of MLL; and (c) a chimeric transcript fusing MLL to the 3'-end of AF9, mimicking the t(9;11)(p22;q23) and expressed 1024-fold higher than the other two. The MLL PNTD, therefore, contributes toward leukemogenesis through simultaneous production of fusion transcripts that are otherwise encoded by three distinct genetic defects.

Absence of the wild-type allele predicts poor prognosis in adult de novo acute myeloid leukemia with normal cytogenetics and the internal tandem duplication of FLT3: a cancer and leukemia group B study.

The FLT3 gene is mutated by an internal tandem duplication (ITD) in 20-25% of adults with acute myeloid leukemia (AML). We studied 82 adults <60 years of age with primary AML and normal cytogenetics, who received uniform high-dose therapy and found FLT3 ITD in 23 (28%) patients. When the 23 FLT3 ITD+ cases were compared with the 59 cases with wild-type (WT) FLT3, disease-free survival (DFS) was inferior (P = 0.03), yet overall survival (OS) was not different (P = 0.14). However, 8 (35%) of 23 FLT3 ITD/+ cases also lacked a FLT3 WT allele (FLT3(ITD-R)) as determined by PCR and loss of heterozygosity. Thus, three genotypic groups were identified: normal FLT3(WT/WT), heterozygous FLT3(ITD/WT), and hemizygous FLT3(ITD/-). DFS and OS were significantly inferior for patients with FLT3(ITD/-) (P = 0.0017 and P = 0.0014, respectively). Although DFS and OS for FLT3(WT/WT) and FLT3(ITD/WT) groups did not differ (P = 0.32 and P = 0.98, respectively), OS of the FLT3(ITD/-) group was worse than the FLT3(WT/WT) (P = 0.0005) and FLT3(ITD/WT) (P = 0.008) groups. We propose a model in which FLT3(ITD/-) represents a dominant positive, gain-of-function mutation providing AML cells with a greater growth advantage compared with cells having the FLT3(WT/WT) or FLT3(ITD/WT) genotypes. In conclusion, we have identified the FLT3(ITD/-) genotype as an adverse prognostic factor in de novo AML with normal cytogenetics. A poor prognosis of the relatively young FLT3(ITD/-) adults (median age, 37 years), despite treatment with current dose-intensive regimens, suggests that new treatment modalities, such as therapy with a FLT3 tyrosine kinase inhibitor, are clearly needed for this group of patients.

UBASH3B/Sts-1-CBL axis regulates myeloid proliferation in human preleukemia induced by AML1-ETO.

The t(8;21) rearrangement, which creates the AML1-ETO fusion protein, represents the most common chromosomal translocation in acute myeloid leukemia (AML). Clinical data suggest that CBL mutations are a frequent event in t(8;21) AML, but the role of CBL in AML1-ETO-induced leukemia has not been investigated. In this study, we demonstrate that CBL mutations collaborate with AML1-ETO to expand human CD34+ cells both in vitro and in a xenograft model. CBL depletion by shRNA also promotes the growth of AML1-ETO cells, demonstrating the inhibitory function of endogenous CBL in t(8;21) AML. Mechanistically, loss of CBL function confers hyper-responsiveness to thrombopoietin and enhances STAT5/AKT/ERK/Src signaling in AML1-ETO cells. Interestingly, we found the protein tyrosine phosphatase UBASH3B/Sts-1, which is known to inhibit CBL function, is upregulated by AML1-ETO through transcriptional and miR-9-mediated regulation. UBASH3B/Sts-1 depletion induces an aberrant pattern of CBL phosphorylation and impairs proliferation in AML1-ETO cells. The growth inhibition caused by UBASH3B/Sts-1 depletion can be rescued by ectopic expression of CBL mutants, suggesting that UBASH3B/Sts-1 supports the growth of AML1-ETO cells partly through modulation of CBL function. Our study reveals a role of CBL in restricting myeloid proliferation of human AML1-ETO-induced leukemia, and identifies UBASH3B/Sts-1 as a potential target for pharmaceutical intervention.

Depsipeptide (FR 901228) promotes histone acetylation, gene transcription, apoptosis and its activity is enhanced by DNA methyltransferase inhibitors in AML1/ETO-positive leukemic cells.

In t(8;21) acute myeloid leukemia (AML), the AML1/ETO fusion protein promotes leukemogenesis by recruiting histone deacetylase (HDAC) and silencing AML1target genes important for hematopoietic differentiation. We hypothesized that depsipeptide (FR901228), a novel HDAC inhibitor evaluated in ongoing clinical trials, restores gene transcription and cell differentiation in AML1/ETO-positive cells. A dose-dependent increase in H3 and H4 histone acetylation was noted in depsipeptide-treated AML1/ETO-positive Kasumi-1 cells and blasts from a patient with t(8;21) AML. Consistent with this biological effect, we also showed a dose-dependent increase in cytotoxicity, expression of IL-3, here used as read-out for silenced AML1-target genes, upregulation of CD11b with other morphologic changes suggestive of partial cell differentiation in Kasumi-1 cells. Some of these biologic effects were also attained in other myeloid leukemia cell lines, suggesting that depsipeptide has differentiation and cytotoxic activity in AML cells, regardless of the underlying genomic abnormality. Notably, the activity of depsipeptide was enhanced by 5-aza-2'-deoxycytidine, a DNA methyltransferase inhibitor (DNMT). These two agents in combination resulted in enhanced histone acetylation, IL-3 expression, and cytotoxicity, suggesting HDAC and DNMT activities as a potential dual target in future therapeutic strategies for AML1/ETO and other molecular subgroups of AML.

Marked declines in human immunodeficiency virus-related mortality in Chicago in women, African Americans, Hispanics, young adults, and injection drug users, from 1995 through 1997.

BACKGROUND: Declines in human immunodeficiency virus (HIV)-related mortality between 1995 and 1996 were seen across the United States but were small to nonexistent among people in marginalized sectors who are most likely to contract HIV and die of its effects. No comprehensive analysis describing HIV-related mortality in 1997 was available. OBJECTIVE: To describe Chicago's HIV-related mortality trends up to and including 1997, with specific attention focused on marginalized populations. METHODS: An analysis of cross-sectional HIV-related mortality data with emphasis on the years 1995 through 1997 was conducted for Chicago, Ill. Numbers, proportions, and rates of declines in HIV-related deaths were examined for the city as a whole and also among those diagnosed at Cook County Hospital, as a proxy for people with very low socioeconomic status. RESULTS: Between 1995 and 1996 there was an overall decline of 19% in HIV-related mortality in Chicago but small or no declines among women, African Americans, Hispanics, injection drug users, and people aged 20 to 29 years and more than 50 years. Between 1995 and 1997 there was an overall decline of 61%. At that time the declines were spread more evenly across diverse groups. There were almost no significant differences between the declines for these groups at Cook County Hospital and in the rest of Chicago. CONCLUSIONS: The HIV-related mortality has fallen dramatically in Chicago since 1995, the year of its maximum. During 1997, declines were seen among all groups. Declines were also seen among the most disenfranchised of the city. Access to care and the new combination therapies are apparently sustaining life for many in Chicago.

Receptor tyrosine kinase Axl is required for resistance of leukemic cells to FLT3-targeted therapy in acute myeloid leukemia.

In acute myeloid leukemia (AML), about 25-30% of patients harbor a constitutively active receptor tyrosine kinase (RTK) FLT3 encoded by a FLT3 allele harboring internal tandem duplication (FLT3-ITD) mutation. The presence of FLT3-ITD correlates with poor prognosis in AML and it makes FLT3 an attractive therapeutic target in AML. Unfortunately, to date small-molecule inhibitors of FLT3 have resulted in only partial and transient clinical responses with residual leukemic blasts resistant to FLT3 inhibitors detected in blood or bone marrow. In this study, we investigated whether the RTK Axl is responsible for resistance of FLT3-ITD(+) AML cells to PKC412 and AC220, FLT3 inhibitors currently under clinical trials for FLT3-ITD(+) AML patients. Upon treatment with PKC412 or AC220, phosphorylation of Axl was significantly enhanced in the FLT3-ITD(+) MV4-11 AML cell line and in primary blasts from a FLT3-ITD(+) AML patient. Consistently, a PKC412-resistant AML cell line and PKC412-resistant primary blasts from FLT3-ITD(+) AML patients had significantly higher levels of constitutively phosphorylated Axl and total Axl when compared with a PKC412-sensitive AML cell line and PKC412-sensitive primary blasts from FLT3-ITD(+) AML patients. We also found that resistance of AML cells against the FLT3 inhibitor PKC412 and AC220 was substantially diminished by the inhibition of Axl via a small-molecule inhibitor TP-0903, a soluble receptor Axl fusion protein Axl-Fc or knockdown of Axl gene expression by shRNA. Collectively, our study suggests that Axl is required for resistance of FLT3-ITD(+) AML cells against the FLT3 inhibitor PKC412 and AC220, and that inhibition of Axl activation may overcome resistance to FLT3-targeted therapy in FLT3-ITD(+) AML.

Prognostic and biologic significance of DNMT3B expression in older patients with cytogenetically normal primary acute myeloid leukemia.

DNMT3B encodes a DNA methyltransferase implicated in aberrant epigenetic changes contributing to leukemogenesis. We tested whether DNMT3B expression, measured by NanoString nCounter assay, associates with outcome, gene and microRNA expression and DNA methylation profiles in 210 older (⩾60 years) adults with primary, cytogenetically normal acute myeloid leukemia (CN-AML). Patients were dichotomized into high versus low expressers using median cut. Outcomes were assessed in the context of known CN-AML prognosticators. Gene and microRNA expression, and DNA methylation profiles were analyzed using microarrays and MethylCap-sequencing, respectively. High DNMT3B expressers had fewer complete remissions (CR; P=0.002) and shorter disease-free (DFS; P=0.02) and overall (OS; P<0.001) survival. In multivariable analyses, high DNMT3B expression remained an independent predictor of lower CR rates (P=0.04) and shorter DFS (P=0.04) and OS (P=0.001). High DNMT3B expression associated with a gene expression profile comprising 363 genes involved in differentiation, proliferation and survival pathways, but with only four differentially expressed microRNAs (miR-133b, miR-148a, miR-122, miR-409-3p) and no differential DNA methylation regions. We conclude that high DNMT3B expression independently associates with adverse outcome in older CN-AML patients. Gene expression analyses suggest that DNMT3B is involved in the modulation of several genes, although the regulatory mechanisms remain to be investigated to devise therapeutic approaches specific for these patients.

Psychopathology in epilepsy: how great is the risk?

The relationship between epilepsy and psychiatric disorder in general and the relationship between epilepsy and psychosis in particular remain controversial issues of long standing. In order to reexamine these central issues concerned with epilepsy/psychopathology relationships, we utilized a Minnesota Multiphasic Personality Inventory (MMPI) sequential diagnostic system to reanalyze 87 published MMPI profiles of patients with epilepsy, other neurological disorders, and chronic physical illnesses encompassing a total of 2786 patients. We found that overall rates of psychopathology were not increased in epilepsy. However, when psychopathology was present, the level of psychosis was greatest in patients with epilepsy. The results are related to the larger literature concerned with the relationship between epilepsy and psychopathology.

Epilepsy in prison: elevated prevalence and no relationship to violence.

The epidemiologic, clinical, and social characteristics of epilepsy were investigated in men entering the Illinois prison system and compared with a matched control group of prisoners without epilepsy. The prevalence of epilepsy was 2.4%, four times higher than the prevalence among men aged 20 to 39 in Rochester, MN. Head trauma was the probable cause of epilepsy among 45% of the prisoners with epilepsy, a much higher percentage than that reported in studies of other populations. In comparison with an age- and race-matched group of prisoners without epilepsy, the epilepsy group was not convicted of more serious or more violent crimes.

Asthma hospitalizations and mortality in Chicago: an epidemiologic overview.

STUDY OBJECTIVES: To characterize the patterns and correlates of asthma hospitalizations and mortality in Chicago. DESIGN: Cross-sectional analysis of discharge data for 1996 and mortality time trend data for the period from 1990 to 1997. SETTING: The city of Chicago, IL, with Cook County, IL, and US data employed for comparisons. POPULATION STUDIED: People who were hospitalized with a primary diagnosis of asthma and people whose underlying cause of death was asthma. INTERVENTIONS: None. MEASUREMENTS AND RESULTS: The 1996 asthma hospitalization rate for Chicago was 42.8 per 10,000, more than twice as high as suburban Chicago or US rates. Medicaid patients were overrepresented. Length of stay was longer for older patients and Medicaid patients. Age-adjusted asthma mortality in Chicago was 4.7 times higher in non-Hispanic blacks than in non-Hispanic whites. The black/white asthma mortality ratio is 2.5:1 for the nation overall. Asthma mortality rates for Hispanics in Chicago were between those of non-Hispanic whites and blacks but have almost doubled during this decade. CONCLUSIONS: The rising asthma mortality and high asthma hospitalization rates in Chicago constitute a significant public health problem. Comorbidities more common in urban environments, such as substance abuse, may play a unique role in determining the distribution of adverse outcomes within Chicago's population. Asthma hospitalizations and deaths may vary in their risk profiles, and this should be taken into account when developing research and intervention strategies.

Do chart reviews and interviews provide the same information about breast and cervical cancer screening?

The National Cancer Institute of the United States has set a goal for the year 2000 that 80-90% of eligible women should have a Pap smear every 3 years and that 80% of women aged 50-70 should receive an annual breast examination and mammogram. Very few studies have examined how we might best measure our progress towards this goal. Specifically, should we employ interview data or data derived from medical records? To respond to this question, data were gathered at two different public health clinics in poor areas of Chicago using both techniques. The interviews estimated significantly higher proportions of women receiving Pap smears, breast examinations, and mammograms in the previous 12-month interval than were estimated from randomly selected medical records. A review of the literature suggests the same pattern exists when other studies using these two data gathering processes are compared. We are thus left with a serious problem, one that must be resolved before we will be able to fully assess our progress in increasing breast and cervical cancer screening.

Sequelae associated with head injuries in patients who were not hospitalized: a follow-up survey.

Data about sequelae associated with head injuries in patients presenting at a suburban hospital but not hospitalized were collected from emergency department medical records and two follow-up telephone interviews. During the study period 669 patients with head injuries were discharged from the emergency department. Of these, 288 were asked to participate in the study, 275 (95%) agreed, and 262 (91%) were eventually contacted. Participants and nonparticipants were compared on six variables and differed significantly only on age--younger patients were more likely to be included. Forty-eight hours after trauma, 52% of the respondents suffered headaches, 14% complained of dizziness, and 13% complained of drowsiness. One week after trauma, the complaints were headaches in 27%, dizziness in 11%, and drowsiness in 9%. Twenty-seven per cent had not resumed normal activity at 48 hours after trauma, and 13% had not at 1 week. Sixty-six per cent of the patients followed the patient instructions regarding head injuries.

The architecture of research in the epilepsy/psychopathology field.

In order to examine how research about the relationship between epilepsy and psychopathology has proceeded, the literature that has appeared since 1966 was reviewed and 78 relevant studies were located. The variables employed in these studies (510 in all) were tabulated to determine if they examined biological variables, social variables or medication variables as etiology of psychopathology in people with epilepsy. The results indicate an overwhelming tendency to omit social variables when searching for etiological factors. It is concluded that such an omission is of theoretical and practical significance and 3 hypotheses are put forward to account for the current distribution of research efforts. Finally, the implications of such a structure of investigation are considered.

Multietiological determinants of psychopathology and social competence in children with epilepsy.

The purpose of this investigation was to inquire into the multietiological determinants of psychopathology and social competence in children with epilepsy. The relationship between behavioral functioning as assessed by the Child Behavior Checklist and a variety of biological, psychosocial, medication and demographic risk factors was investigated in a sample of 183 children with epilepsy aged 6-16. Several risk factors were found to be related to each behavioral measure. The results are discussed both in terms of their implications for models of psychopathology in epilepsy as well as their relationship to previous findings in the epilepsy/psychopathology field.

Psychopathology in epilepsy. The role of psychology in altering paradigms of research, treatment, and prevention.

This article offers a brief review of the literature examining epilepsy and abnormal behavior. The generally accepted concept of epilepsy as a high psychiatric risk disorder is reexamined in light of research that has investigated the influence of selection bias. A conceptual model that organizes the known or postulated risk variables (neurological, psychosocial, and medication) is presented, with emphasis on psychology's potential contribution.