Patterns of adversity and post-traumatic stress among children adopted from care.

BACKGROUND: Children adopted from care are more likely to have experienced early adversity, but little is known about the impact of early adversity on later post-traumatic stress (PTS) symptoms. OBJECTIVE: To investigate sub-groups of adversity in a sample of adopted children and examine the association with later PTS symptoms. PARTICIPANTS AND SETTING: A study of British children adopted from care using social worker records (N = 374) and questionnaire-based longitudinal study of n = 58 children over 4-years post adoptive placement. METHODS: We used latent class analysis to identify subgroups of children based on commonalities in perinatal and postnatal adversity experienced prior to adoption and examined differences in PTS symptoms at 4-years post-placement between subgroups. RESULTS: Nearly one in five (19 %) children were in the clinical or borderline ranges for symptoms of PTS arousal, 14 % for PTS avoidance and 8 % for PTS intrusion. The 5-class solution fitted the data best, with one class characterized by children with a low probability of experiencing any adversity, one perinatal adversity class and three classes capturing different patterns of adversity. The multiple complex adversity class involving both perinatal and postnatal adversity had significantly higher symptoms of PTS avoidance and arousal than other sub-groups. CONCLUSIONS: The prevalence and complexity of PTS symptoms among adoptive children highlights the need for effective interventions considering different profiles of early adversity.

Palliative Care: Critical Concepts for the Geropsychiatrist.

Psychiatrists can make a significant contribution to improving quality end-of-life care for psychiatric patients, beyond managing their psychiatric and psychological conditions. Geriatric psychiatrists can build expertise in enhancing end-of-life care when caring for older adults with serious illnesses and their families, given the biopsychosociospiritual approach that significantly overlaps with palliative and hospice care approaches. To effectively add quality to end-of-life care, it is essential for psychiatrists to understand the core principles and practices of palliative and hospice care, learn basic symptom management skills, and hone the ability to have crucial conversations regarding prognosis and advance care planning. Also important is recognizing when to refer to hospice and palliative medicine subspecialists. This article provides an overview of palliative and hospice care, uses a case study to illustrate components of palliative and hospice care relevant to geriatric psychiatry practice, and comments on considerations pertinent to the coronavirus disease 2019 (COVID-19) pandemic.

Characterization of a cis-acting regulatory element which silences expression of the class II-A beta gene in epithelium.

Class II major histocompatibility complex (MHC) genes encode for alpha/beta chain pairs that are constitutively expressed principally on mature B cells and dendritic cells in mice. These gene products are easily induced on macrophages with cytokines, and may also aberrantly appear on the surface of epithelium during immune injury. The appearance of class II determinants in parenchymal tissue potentially renders these somatic cells capable of antigen presentation to circulating CD4+ T lymphocytes, and their absence may be protective for normal tissues expressing self-antigens. The low surface class II expression observed on parenchymal cells generally correlates with low levels of mRNA, suggesting that transcription rate is a major element in class II regulation. To understand the transcriptional mechanism maintaining low basal surface expression of class II in somatic cells, we transiently transfected mini-gene reporter constructs to study the regulation of the murine A beta promoter in a cultured renal epithelial cell line. We describe here a negative cis-acting regulatory region located between -552 and -489 bp upstream of the A beta cap site that silences the transcriptional activity of the A beta promoter in epithelial cells in an orientation-dependent manner, and is also able to silence a heterologous promoter. This region is not active in class II-expressing B cells (BAL-17) in culture, but is functional in two other murine class II-negative cell lines, fibroblasts and thymoma T cells. Using competition electrophoretic mobility shift assays, we have localized the core protein binding site within this region to an 8-10-bp response element, designated A beta NRE, at -543 to -534 bp. A nuclear extract from BAL-17 cells does not bind to this element. Mutation of this site abrogates the transcriptional silencing activity of the region. We conclude that the transcription of class II-A beta in parenchymal cells, and some lymphocytes, can be actively repressed by an upstream silencing element.

Transgender health education for medical students.

BACKGROUND: Transgender individuals face numerous health disparities and report negative experiences with health care providers related to their gender identity. Significant gaps in medical education regarding transgender health persist despite calls for increased sexual and gender minority content. The purpose of this student-led study was to assess the effectiveness of a half-day educational intervention on first- and second-year medical students' attitudes and knowledge of transgender health. METHODS: Students and faculty members collaborated to develop an educational session on transgender health. This content was presented to first- and second-year medical students at Integrated Grand Rounds, a pedagogical method in which basic science and clinical faculty members co-present didactic content interspersed between live patient interviews and student-led small group discussions. Student participants (n = 138) completed voluntary 9-item pre- and post-session surveys assessing comfort with and knowledge of transgender medicine. RESULTS: Students' comfort with and perceived knowledge about transgender patients increased significantly between pre- and post-test. Students' knowledge of transgender medicine standards of care also improved, though not all items reached significance. DISCUSSION: A half-day educational intervention improved many facets of medical students' attitudes and knowledge about transgender patients. The significant disparities in physical health, mental health and access to care currently experienced by transgender persons in the United States warrants the continued testing and refinement of educational interventions for future and practising providers. Students' comfort with transgender patients increased significantly between pre- and post-test.

FGF2-FGFR1 signaling regulates release of Leukemia-Protective exosomes from bone marrow stromal cells.

Protective signaling from the leukemia microenvironment leads to leukemia cell persistence, development of resistance, and disease relapse. Here, we demonstrate that fibroblast growth factor 2 (FGF2) from bone marrow stromal cells is secreted in exosomes, which are subsequently endocytosed by leukemia cells, and protect leukemia cells from tyrosine kinase inhibitors (TKIs). Expression of FGF2 and its receptor, FGFR1, are both increased in a subset of stromal cell lines and primary AML stroma; and increased FGF2/FGFR1 signaling is associated with increased exosome secretion. FGFR inhibition (or gene silencing) interrupts stromal autocrine growth and significantly decreases secretion of FGF2-containing exosomes, resulting in less stromal protection of leukemia cells. Likewise, Fgf2 -/- mice transplanted with retroviral BCR-ABL leukemia survive significantly longer than their +/+ counterparts when treated with TKI. Thus, inhibition of FGFR can modulate stromal function, reduce exosome secretion, and may be a therapeutic option to overcome resistance to TKIs. Editorial note: This article has been through an editorial process in which the authors decide how to respond to the issues raised during peer review. The Reviewing Editor's assessment is that all the issues have been addressed (see decision letter).

Multiple elements in the upstream glucokinase promoter contribute to transcription in insulinoma cells.

beta-cell type-specific expression of the upstream glucokinase promoter was studied by transfection of fusion genes and analysis of DNA-protein interactions. A construct containing 1,000 bp of 5'-flanking DNA was efficiently expressed in HIT M2.2.2 cells, a beta-cell-derived line that makes both insulin and glucokinase, but not in NIH 3T3 cells, a heterologous cell line. In a series of 5' deletion mutations between bases -1000 and -100 (relative to a base previously designated +1), efficient expression in HIT cells was maintained until -280 bp, after which transcription decreased in a stepwise manner. The sequences between -180 and -1 bp contributing to transcriptional activity in HIT cells were identified by studying 28 block transversion mutants that spanned this region in 10-bp steps. Two mutations reduced transcription 10-fold or more, while six reduced transcription between 3- and 10-fold. Three mutationally sensitive regions of this promoter were found to bind to a factor that was expressed preferentially in pancreatic islet beta cells. The binding sites, designated upstream promoter elements (UPEs), shared a consensus sequence of CAT(T/C)A(C/G). Methylation of adenine and guanine residues within this sequence prevented binding of the beta-cell factor, as did mutations at positions 2, 3, and 5. Analysis of nuclear extracts from different cell lines identified UPE-binding activity in HIT M2.2.2 and beta-TC-3 cells but not in AtT-20, NIH 3T3, or HeLa cells; the possibility of a greatly reduced amount in alpha-TC-6 cells could not be excluded. UV laser cross-linking experiments supported the beta-cell type expression of this factor and showed it to be approximately 50 kDa in size. Gel mobility shift competition experiments showed that this beta-cell factor is the same that binds to similar elements, termed CT boxes, in the insulin promoter. Thus, a role for these elements (UPEs or CT boxes), and the beta-cell factor that binds to them, in determining the expression of genes in the beta cells of pancreatic islets is suggested.

BZP, a novel serum-responsive zinc finger protein that inhibits gene transcription.

We report the fortuitous isolation of cDNA clones encoding a novel zinc finger DNA-binding protein termed BZP. The protein encoded is 114 kDa and contains eight zinc finger motifs, seven of which are present in two clusters at opposite ends of the molecule. Both finger clusters bound to the 9-bp sequence AAAGGTGCA with apparent Kds of approximately 2.5 nM. Two of the finger motifs within the amino- and carboxy-terminal finger clusters share 63% amino acid identity. BZP inhibited transcription of the herpes simplex virus thymidine kinase promoter when copies of the 9-bp target motif were linked in cis, suggesting that it functions as a transcriptional repressor. BZP mRNA and immunoreactivity were detected in several established cell lines but were most abundant in hamster insulinoma (HIT) cells, the parental source of the cDNAs. In mouse tissues, BZP mRNA and immunoreactivity were identified in cells of the endocrine pancreas, anterior pituitary, and central nervous system. Interestingly, in HIT cells proliferating in culture, BZP immunoreactivity was predominately nuclear in location, whereas it was usually located in the cytoplasm in most neural and neuroendocrine tissues. Serum deprivation of HIT cells caused BZP immunoreactivity to become predominantly cytoplasmic in location and attenuated its inhibitory effect on transcription, thereby suggesting that the both the subcellular location and the function of this protein are modulated by factors in serum.

Phosphoenolpyruvate carboxykinase is necessary for the integration of hepatic energy metabolism.

We used an allelogenic Cre/loxP gene targeting strategy in mice to determine the role of cytosolic phosphoenolpyruvate carboxykinase (PEPCK) in hepatic energy metabolism. Mice that lack this enzyme die within 3 days of birth, while mice with at least a 90% global reduction of PEPCK, or a liver-specific knockout of PEPCK, are viable. Surprisingly, in both cases these animals remain euglycemic after a 24-h fast. However, mice without hepatic PEPCK develop hepatic steatosis after fasting despite up-regulation of a variety of genes encoding free fatty acid-oxidizing enzymes. Also, marked alterations in the expression of hepatic genes involved in energy metabolism occur in the absence of any changes in plasma hormone concentrations. Given that a ninefold elevation of the hepatic malate concentration occurs in the liver-specific PEPCK knockout mice, we suggest that one or more intermediary metabolites may directly regulate expression of the affected genes. Thus, hepatic PEPCK may function more as an integrator of hepatic energy metabolism than as a determinant of gluconeogenesis.

Development of a cryogenically cooled platform for the Magnetized Liner Inertial Fusion (MagLIF) Program.

A cryogenically cooled hardware platform has been developed and commissioned on the Z Facility at Sandia National Laboratories in support of the Magnetized Liner Inertial Fusion (MagLIF) Program. MagLIF is a magneto-inertial fusion concept that employs a magnetically imploded metallic tube (liner) to compress and inertially confine premagnetized and preheated fusion fuel. The fuel is preheated using a ∼2 kJ laser that must pass through a ∼1.5-3.5-µm-thick polyimide "window" at the target's laser entrance hole (LEH). As the terawatt-class laser interacts with the dense window, laser plasma instabilities (LPIs) can develop, which reduce the preheat energy delivered to the fuel, initiate fuel contamination, and degrade target performance. Cryogenically cooled targets increase the parameter space accessible to MagLIF target designs by allowing nearly 10 times thinner windows to be used for any accessible gas density. Thinner LEH windows reduce the deleterious effects of difficult to model LPIs. The Z Facility's cryogenic infrastructure has been significantly altered to enable compatibility with the premagnetization and fuel preheat stages of MagLIF. The MagLIF cryostat brings the liquid helium coolant directly to the target via an electrically resistive conduit. This design maximizes cooling power while allowing rapid diffusion of the axial magnetic field supplied by external Helmholtz-like coils. A variety of techniques have been developed to mitigate the accumulation of ice from vacuum chamber contaminants on the cooled LEH window, as even a few hundred nanometers of ice would impact laser energy coupling to the fuel region. The MagLIF cryostat has demonstrated compatibility with the premagnetization and preheat stages of MagLIF and the ability to cool targets to liquid deuterium temperatures in approximately 5 min.

Selective effects of nonionic detergent and salt solutions in dissolving nuclear envelope protein.?20U.

Protein has been selectively extracted from isolated chicken erythrocyte nuclear envelope by (1) dilute MgCl2/Triton X-100 followed by (2) concentrated MgCl2/Triton X-100 solutions. Certain proteins appear to be selectively dissolved in the first solvent and may occur in the nuclear envelope primarily as lipoproteins. Among the proteins insoluble in the low MgCl2/Triton X-100 wash, as well as in 500 mM MgCl2 without Triton previously used in the preparation of the envelope fraction, the quantitatively major polypeptides dissolve in a combination of high MgCl2 and Triton X-100. Further, much of this dissolved protein precipitates when the MgCl2 concentration is lowered by dialysis. The insolubility of these proteins appears to result from a combination of ionic and hydrophobic interactions and may explain the resistance of nuclei to various manipulative procedures including nonionic detergent washes. The procedures described provide a route for gently and selectively dissolving representative proteins from the nuclear envelope lipoprotein matrix and from the envelope "residual" protein.

Characterization of the Pal motifs in the upstream glucokinase promoter: binding of a cell type-specific protein complex correlates with transcriptional activation.

The upstream glucokinase (GK) promoter is expressed specifically in several different neural/neuroendocrine (NE) cell types, including the pancreatic beta-cell and pituitary corticotrope. Previously, a mutational and evolutionary analysis of this promoter identified two identical 9-bp motifs (TGGTCACCA) termed Pal-1 and Pal-2 that are essential for high level expression in HIT M2.2.2 cells, an insulinoma cell line. Here we show that these motifs are also necessary for efficient expression in AtT-20 cells, a corticotrope-derived cell line, and that proteins from both NE and non-NE cells bind to the Pal motifs, although the DNA-protein complexes differ by cell type. Complexes formed using nuclear extracts from NE cells contained an extra NE cell-specific band and differed in the relative abundance of two other bands when compared with non-NE cells, UV laser cross-linking experiments further supported the cell-specific binding of two proteins, 110 and 150 kDa in size, to these motifs. The presence or absence of the NE-specific band correlates with transcription of GK promoter fusion gene constructs, suggesting a key role for this protein in determining the cell-specific expression of GK. The Pal motifs themselves do not function as enhancers but seem to be essential components of a larger transcriptional regulatory domain that is active only in certain NE cells. Together, these studies suggest that the NE cell-specific expression of the upstream GK promoter involves the formation of a distinct protein complex on the two Pal motifs.

Effects of acute tryptophan depletion on raphé functional connectivity in depression.

Depression remains a great societal burden and a major treatment challenge. Most antidepressant medications target serotonergic raphé nuclei. Acute tryptophan depletion (ATD) modulates serotonin function. To better understand the raphé's role in mood networks, we studied raphé functional connectivity in depression. Fifteen depressed patients were treated with sertraline for 12 weeks and scanned during ATD and sham conditions. Based on our previous findings in a separate cohort, resting state MRI functional connectivity between raphé and other depression-related regions (ROIs) was analyzed in narrow frequency bands. ATD decreased raphé functional connectivity with the bilateral thalamus within 0.025-0.05 Hz, and also decreased raphé functional connectivity with the right pregenual anterior cingulate cortex within 0.05-0.1 Hz. Using the control broadband filter 0.01-0.1 Hz, no significant differences in raphé-ROI functional connectivity were observed. Post-hoc analysis by remission status suggested increased raphé functional connectivity with left pregenual anterior cingulate cortex in remitters (n=10) and decreased raphé functional connectivity with left thalamus in non-remitters (n=5), both within 0.025-0.05 Hz. Reducing serotonin function appears to alter coordination of these mood-related networks in specific, low frequency ranges. For examination of effects of reduced serotonin function on mood-related networks, specific low frequency BOLD fMRI signals can identify regions implicated in neural circuitry and may enable clinically-relevant interpretation of functional connectivity measures. The biological significance of these low frequency signals detected in the raphé merits further study.

Adipose tissue deposition in sleep apnea.

To determine whether adipose tissue is deposited in the neck adjacent to the upper airway in patients with obstructive sleep apnea (OSA), we studied 21 subjects with OSA and nine without OSA using magnetic resonance imaging with a T-1 weighted spin echo sequence and polysomnography. We observed that patients with OSA had a larger volume of adipose tissue adjacent to their upper airway than did subjects without OSA.

Isolation and characterization of the mouse cytosolic phosphoenolpyruvate carboxykinase (GTP) gene: evidence for tissue-specific hypersensitive sites.

A 72 kilobase pair DNA fragment that contains the mouse phosphoenolpyruvate carboxykinase (PEPCK) gene locus, pck1, was isolated from a genomic bacterial artificial chromosome library. The region from approximately -5.5 to +6.6 kilobase pairs relative to the pck1 transcription start site was sequenced and exhibits a high degree of homology to the rat and human genes. Additionally, the chromatin structure of the PEPCK gene in mouse liver resembles that seen in rat. Backcross panel analysis of a microsatellite sequence confirms that the gene is located on chromosome 2. Hypersensitive site analysis was performed on nuclei isolated from the adipocyte cell line 3T3-F442A in the preadipose and adipose states. Several hypersensitive sites are present in the undifferentiated 3T3-F442A cells, before PEPCK mRNA is detected. The same sites are present after differentiation, however, the sensitivity of mHS 3 increases relative to the others. We conclude that the chromatin is open in 3T3-F442A cells and that factors are able to bind in the undifferentiated state but that something else is required for transcription.

A procedure for purifying low-abundance protein components from the brain cytoskeleton-nuclear matrix fraction.

We describe a preparative procedure for low-abundance proteins of the cytoskeleton-nuclear matrix fraction from frozen bovine brain. Strigent centrifugation and washing conditions in the preparation of the cytoskeleton-nuclear matrix fraction are avoided to minimize loss of nuclear material. A recently described horizontal isoelectric focusing column, which tolerates appreciable precipitation, is used. In concert with selection of urea concentration and temperature, this isoelectric focusing apparatus provides a new approach to the fractionation of this complex, relatively insoluble mixture of proteins and other components. In addition, a heated, sodium dodecyl sulfate-sizing column has been utilized in order to eliminate interactions between the desired low abundance proteins and more abundant contaminating proteins. Together these procedures purify a specific low-abundance protein sufficiently to be detected by Coomassie blue staining in two-dimensional gels. The methods are robust and can be applied to multiple, relatively large brain samples (150 g of crude grey matter per batch); thus they should facilitate partial peptide sequencing for brain proteins of this operational class.

Supersensitivity to the reinforcing effects of cocaine following 6-hydroxydopamine lesions to the medial prefrontal cortex in rats.

The effects of neurotoxic lesions to the medial prefrontal cortex on both the acquisition and maintenance of intravenous cocaine self-administration were examined. In one experiment, acquisition of intravenous cocaine self-administration (0.25, 0.5 or 1.0 mg/kg/infusion) was measured in separate groups of rats 14 days following either a sham or 6-hydroxydopamine lesion to the medial prefrontal cortex. For sham rats, the 1.0 and 0.5 mg/kg dose supported reliable self-administration as indicated by discriminative responding. These rats reliably chose a lever that resulted in the delivery of these doses of cocaine over an inactive lever. Reinforced response rates were reduced when 0.25 mg/kg was the available dose and there was a loss of discriminative responding for some of the rats suggesting that it was close to threshold for self-administration. For rats that sustained a 70% depletion of dopamine in the medial prefrontal cortex, the dose-response curve was an inverse function across the entire dose range tested. In contrast to the data from the control rats, lesioned rats had a high rate of reinforced responses and demonstrated good discrimination for all doses including 0.25 mg/kg/infusion, suggesting a supersensitive response to the initial reward effect of cocaine. Another group of rats was first screened for reliable cocaine self-administration (0.5 mg/kg/infusion) and then subjected to either the prefrontal cortical 6-hydroxydopamine or sham lesion. Dose-response curves for cocaine self-administration were compared 14 days following the infusions. The lesioned rats responded reliably for low doses of cocaine that were unable to maintain responding in sham rats. These data support the hypothesis that the medial prefrontal cortex plays an important role in cocaine self-administration.

A lead-associated nuclear protein which increases in maturing brain and in differentiating neuroblastoma 2A cells exposed to cyclic AMP-elevating agents.

The lead-associated nuclear protein, p32/6.3, increases significantly in the postnatally developing rat cerebral cortex (Egle and Shelton, J. Biol. Chem., 261 (1986) 2294-2298). In the present study, this increase has been identified with late development of the cerebral cortex or forebrain because p32/6.3 reached adult levels 10 to 14 days after birth in guinea pig (a precocial animal) and after hatching in chicken. Comparison with other developmental processes indicates that p32/6.3 reaches adult levels just before or during the period of synapse maturation. Thus p32/6.3 may prove useful as a biochemical indicator of nuclear maturation in this period. The developmental regulation of p32/6.3 was further studied in mouse neuroblastoma 2a (Nb2a) cells. In vitro induction of differentiation of Nb2a cells by serum withdrawal from the culture medium increased p32/6.3, implicating p32/6.3 with differentiating neurons. This association was further strengthened when treatment of the Nb2a cells for 24 h with dibutyryl cAMP (1-5 mM), papaverine (5-12.5 micrograms/ml) or 3-isobutyl-1-methylxanthine (IBMX; 50-250 microM) increased the abundance of p32/6.3 1.5- to 3-fold more than serum withdrawal alone. 8-Bromo-cAMP (2-4 mM), N6-benzoyl cAMP (4 mM) and forskolin (10 microM) also increased the abundance of p32/6.3 in Nb2a cells, arguing that cAMP is involved in p32/6.3 regulation. These results, in conjunction with the postnatal increase of p32/6.3 in cerebral cortex, suggest a relationship between p32/6.3 levels and neuronal maturation.

Ovum retention in the human.

Two cases of ovum retention occurring in postovulatory follicles are described. The ova were recovered at laparoscopy by aspiration of decompressed ovulatory follicles, one during a natural cycle and the other following a programmed clomiphene/human chorionic gonadotropin cycle. Each patient had a normal luteal phase with an increased progesterone level indicative of ovulation. The implications of these findings and their relevance to human fertility studies are discussed.