Functional asymmetry of the human brain.

Verbal and nonverbal memorization skills were tested before and after electroconvulsive shocks to the left, right, or both cerebral hemispheres of neurologically normal patients. As predicted, decrements for the left-hemisphere-shocked group were larger on the verbal than nonverbal tasks, while the reverse was true for the right-hemisphere-shocked group. Largest decrements on both tasks were shown by the bilaterally shocked group.

Mating and pregnancy can occur in genetically hypogonadal mice with preoptic area brain grafts.

Adult female hypogonadal mice, in whom hypogonadism is secondary to a genetic deficiency in hypothalamic gonadotropin-releasing hormone (GnRH), are infertile. Mating, pregnancy, and delivery of healthy litters were achieved after transplantation of normal fetal preoptic area tissue, a major site of GnRH-containing cell bodies, into the third ventricle of adult female hypogonadal mice. Immunocytochemistry revealed GnRH-containing neurons in the grafts and GnRH-containing processes extending to the lateral median eminence of the host brains.

Gene for familial psoriasis susceptibility mapped to the distal end of human chromosome 17q.

A gene involved in psoriasis susceptibility was localized to the distal region of human chromosome 17q as a result of a genome-wide linkage analysis with polymorphic microsatellites and eight multiply affected psoriasis kindreds. In the family which showed the strongest evidence for linkage, the recombination fraction between a psoriasis susceptibility locus and D17S784 was 0.04 with a maximum two-point lod score of 5.33. There was also evidence for genetic heterogeneity and although none of the linked families showed any association with HLA-Cw6, two unlinked families showed weak levels of association. This study demonstrates that in some families, psoriasis susceptibility is due to variation at a single major genetic locus other than the human lymphocyte antigen locus.

Eliciting the impacts of cellular noise on metabolic trade-offs by quantitative mass imaging.

Optimal metabolic trade-offs between growth and productivity are key constraints in strain optimization by metabolic engineering; however, how cellular noise impacts these trade-offs and drives the emergence of subpopulations with distinct resource allocation strategies, remains largely unknown. Here, we introduce a single-cell strategy for quantifying the trade-offs between triacylglycerol production and growth in the oleaginous microorganism Yarrowia lipolytica. The strategy relies on high-throughput quantitative-phase imaging and, enabled by nanoscale secondary ion mass spectrometry analyses and dedicated image processing, allows us to image how resources are partitioned between growth and productivity. Enhanced precision over population-averaging biotechnologies and conventional microscopy demonstrates how cellular noise impacts growth and productivity differently. As such, subpopulations with distinct metabolic trade-offs emerge, with notable impacts on strain performance and robustness. By quantifying the self-degradation of cytosolic macromolecules under nutrient-limiting conditions, we discover the cell-to-cell heterogeneity in protein and fatty-acid recycling, unmasking a potential bet-hedging strategy under starvation.

Cyclin D- and E-dependent kinases and the p57(KIP2) inhibitor: cooperative interactions in vivo.

This study examines in vivo the role and functional interrelationships of components regulating exit from the G1 resting phase into the DNA synthetic (S) phase of the cell cycle. Our approach made use of several key experimental attributes of the developing mouse lens, namely its strong dependence on pRb in maintenance of the postmitotic state, the down-regulation of cyclins D and E and up-regulation of the p57(KIP2) inhibitor in the postmitotic lens fiber cell compartment, and the ability to target transgene expression to this compartment. These attributes provide an ideal in vivo context in which to examine the consequences of forced cyclin expression and/or of loss of p57(KIP2) inhibitor function in a cellular compartment that permits an accurate quantitation of cellular proliferation and apoptosis rates in situ. Here, we demonstrate that, despite substantial overlap in cyclin transgene expression levels, D-type and E cyclins exhibited clear functional differences in promoting entry into S phase. In general, forced expression of the D-type cyclins was more efficient than cyclin E in driving lens fiber cells into S phase. In the case of cyclins D1 and D2, ectopic proliferation required their enhanced nuclear localization through CDK4 coexpression. High nuclear levels of cyclin E and CDK2, while not sufficient to promote efficient exit from G1, did act synergistically with ectopic cyclin D/CDK4. The functional differences between D-type and E cyclins was most evident in the p57(KIP2)-deficient lens wherein cyclin D overexpression induced a rate of proliferation equivalent to that of the pRb null lens, while overexpression of cyclin E did not increase the rate of proliferation over that induced by the loss of p57(KIP2) function. These in vivo analyses provide strong biological support for the prevailing view that the antecedent actions of cyclin D/CDK4 act cooperatively with cyclin E/CDK2 and antagonistically with p57(KIP2) to regulate the G1/S transition in a cell type highly dependent upon pRb.

Mast cells in the brain: evidence and functional significance.

For the past two decades the brain has been considered to be an immune-privileged site that excludes circulating cells from the parenchyma. New evidence indicates that some hematocytes reside in the brain, while others traffic through it. Mast cells belong to both of these functional types. Moreover, the appearance of mast cells in the CNS can be triggered behaviorally. After a brief period of courtship, for example, there is a marked increase in mast cells in the medial habenula of sexually active doves compared with controls. Exposure to gonadal steroids that occur endogenously or that are administered exogenously increases both the number of mast cells and their state of activation in the brain. These results show that hematopoietic cells can provide targeted delivery of neuromodulators to specific regions of the brain, thereby influencing neural-endocrine interactions.

Abnormal methylation of the calcitonin gene in human colonic neoplasms.

Earlier studies of the methylation status of total genomic DNA and of specific genes have demonstrated, predominantly, hypomethylation in human neoplasms. However, we have recently documented the presence of new sites of methylation in the calcitonin gene in human lymphomas (100%), small cell lung carcinomas (92%), and acute myeloid leukemias (95%). We now report that these same novel calcitonin gene methylation sites are also a feature of DNA from human colonic adenomas (13 out of 14 studied), colon carcinomas (4/13), and established colon carcinoma cell lines (18/19), despite the presence of overall genomic DNA hypomethylation in these neoplasms. The data provide further evidence that regional increases in DNA methylation, like gene hypomethylation, occur in benign colonic neoplasms prior to malignant transformation. The fact that abnormalities of calcitonin gene methylation are less frequent in DNA from human colonic carcinomas than from adenomas and colon carcinoma cell culture lines is of special interest. This finding suggests that a more heterogeneous population of cells is present in the carcinomas and that the calcitonin gene hypermethylation may be inherent to cells which are initially selected for growth in culture or are capable of prolonged survival under culture conditions.

Excessive coupling of the salience network with intrinsic neurocognitive brain networks during rectal distension in adolescents with irritable bowel syndrome: a preliminary report.

BACKGROUND: The neural network mechanisms underlying visceral hypersensitivity in irritable bowel syndrome (IBS) are incompletely understood. It has been proposed that an intrinsic salience network plays an important role in chronic pain and IBS symptoms. Using neuroimaging, we examined brain responses to rectal distension in adolescent IBS patients, focusing on determining the alteration of salience network integrity in IBS and its functional implications in current theoretical frameworks. We hypothesized that (i) brain responses to visceral stimulation in adolescents are similar to those in adults, and (ii) IBS is associated with an altered salience network interaction with other neurocognitive networks, particularly the default mode network (DMN) and executive control network (ECN), as predicted by the theoretical models. METHODS: Irritable bowel syndrome patients and controls received subliminal and liminal rectal distension during imaging. Stimulus-induced brain activations were determined. Salience network integrity was evaluated by the functional connectivity of its seed regions activated by rectal distension in the insular and cingulate cortices. KEY RESULTS: Compared with controls, IBS patients demonstrated greater activation to rectal distension in neural structures of the homeostatic afferent and emotional arousal networks, especially the anterior cingulate and insular cortices. Greater brain responses to liminal vs subliminal distension were observed in both groups. Particularly, IBS is uniquely associated with an excessive coupling of the salience network with the DMN and ECN in their key frontal and parietal node areas. CONCLUSIONS & INFERENCES: Our study provided consistent evidence supporting the theoretical predictions of altered salience network functioning as a neuropathological mechanism of IBS symptoms.

Mast cells migrate from blood to brain.

It is well established that mast cells (MCs) occur within the CNS of many species. Furthermore, their numbers can increase rapidly in adults in response to altered physiological conditions. In this study we found that early postpartum rats had significantly more mast cells in the thalamus than virgin controls. Evidence from semithin sections from these females suggested that mast cells were transiting across the medium-sized blood vessels. We hypothesized that the increases in mast cell number were caused by their migration into the neural parenchyma. To this end, we purified rat peritoneal mast cells, labeled them with the vital dyes PKH26 or CellTracker Green, and injected them into host animals. One hour after injection, dye-filled cells, containing either histamine or serotonin (mediators stored in mast cells), were located close to thalamic blood vessels. Injected cells represented approximately 2-20% of the total mast cell population in this brain region. Scanning confocal microscopy confirmed that the biogenic amine and the vital dye occurred in the same cell. To determine whether the donor mast cells were within the blood-brain barrier, we studied the localization of dye-marked donor cells and either Factor VIII, a component of endothelial basal laminae, or glial fibrillary acidic protein, the intermediate filament found in astrocytes. Serial section reconstructions of confocal images demonstrated that the mast cells were deep to the basal lamina, in nests of glial processes. This is the first demonstration that mast cells can rapidly penetrate brain blood vessels, and this may account for the rapid increases in mast cell populations after physiological manipulations.

Pulmonary embolism after cardiac surgery.

OBJECTIVES: We examined the incidence of pulmonary embolism after cardiac surgery. BACKGROUND: Because venous thromboembolism is considered to be an uncommon complication after cardiac surgery, its incidence was documented in a consecutive series of 1,033 patients who underwent cardiac surgery over a 5-year period. METHODS: Parallel cohorts of patients in a tertiary referral center were evaluated and the incidence of pulmonary embolism was compared in subgroups of patients undergoing coronary bypass surgery, valve surgery and combined procedures. RESULTS: Pulmonary embolism developed in 33 (3.2%) of the 1,033 cardiac surgical patients, within 2 weeks of a coronary bypass operation in most; it did not develop in any patient who had isolated valve replacement surgery (p < 0.05). The diagnosis of pulmonary embolism was established by pulmonary angiography in 24 patients, ventilation/perfusion lung scan in 3, postmortem examination in 5 and clinical examination in 1 patient. Important risk factors for pulmonary embolism included prolonged postoperative recovery, obesity and hyperlipidemia. The mortality rate was 18.7% in patients with in contrast to 3.3% in those without pulmonary embolism (p < 0.01). CONCLUSIONS: Although pulmonary embolism is rare after isolated valve replacement, it is not an uncommon complication after coronary bypass surgery.

Hypothalamic transplantation repair of reproductive defects in hypogonadal mice.

The defect of the hypogonadal mouse, resulting in infantile reproductive organs and severely reduced gonadotropin levels, is due to a truncation of the gene encoding for preprogonadotropinreleasing hormone. The hypogonadal mouse bearing a graft containing normal gonadotropin-releasing hormone neurons may show testicular development, seminal vesicle growth, and increased gonadotropin production in males. Normalization of gonadotropin levels in females is frequently associated with the capacity for a reflex ovulation followed by pregnancy and bearing of live young. All of these phenomena are dependent on the outgrowth of gonadotropin-releasing hormone axons from the graft to the host median eminence and the hypophysial portal capillaries.

Methotrexate-induced cirrhosis requiring liver transplantation in three patients with psoriasis. A word of caution in light of the expanding use of this 'steroid-sparing' agent.

Methotrexate has been used for many years to treat refractory psoriasis. Three cases of methotrexate-induced cirrhosis requiring orthotopic liver transplantation are presented to emphasize the importance of strict adherence to published criteria for patient selection, monitoring of cumulative drug dosages, and the performance of serial liver biopsies. Each patient had been treated with long-term methotrexate therapy (cumulative doses far in excess of 1.5 g) without undergoing serial liver biopsies, contrary to well-established treatment guidelines. Caution must be exercised in using methotrexate as a steroid-sparing agent in the treatment of inflammatory diseases because of its potential to cause severe hepatotoxic effects with long-term usage and cumulative doses above 1.5 g. Patients easily become psychologically dependent on the drug, and physicians need to guard against the false sense of security engendered by normal results on liver function studies.

Direct innervation of GnRH neurons by encephalic photoreceptors in birds.

In nonmammalian vertebrates, photic cues that regulate the timing of seasonal reproductive cyclicity are detected by nonretinal, nonpineal deep brain photoreceptors. It has long been assumed that the underlying mechanism involves the transmission of photic information from the photoreceptor to a circadian system, and thence to the reproductive axis. An alternative hypothesis is that there is direct communication between the brain photoreceptor and the reproductive axis. In the present study, light and confocal microscopy reveal that gonadotropin releasing hormone (GnRH) neurons and processes are scattered among photoreceptor cells (identified by their opsin-immunoreactivity) in the lateral septum (SL). In the median eminence (ME), opsin and GnRH immunoreactive fibers overlap extensively. Single and double label ultrastructural immunocytochemistry indicate that in the SL and preoptic area (POA), opsin positive terminals form axo-dendritic synapses onto GnRH dendrites. In the ME, opsin and GnRH terminals lie adjacent to each other, make contact with tanycytes, or terminate on the hypophyseal portal capillaries. These results reveal thatbrain photoreceptors communicate directly with GnRH-neurons; this represents a means by which photoperiodic information reaches the reproductive axis.

Origins of Cell-to-Cell Bioprocessing Diversity and Implications of the Extracellular Environment Revealed at the Single-Cell Level.

Bioprocess limitations imposed by microbial cell-to-cell phenotypic diversity remain poorly understood. To address this, we investigated the origins of such culture diversity during lipid production and assessed the impact of the fermentation microenvironment. We measured the single-cell lipid production dynamics in a time-invariant microfluidic environment and discovered that production is not monotonic, but rather sporadic with time. To characterize this, we introduce bioprocessing noise and identify its epigenetic origins. We linked such intracellular production fluctuations with cell-to-cell productivity diversity in culture. This unmasked the phenotypic diversity amplification by the culture microenvironment, a critical parameter in strain engineering as well as metabolic disease treatment.

Characterization of skin-infiltrating lymphocytes in patients with psoriasis.

In this study, skin-infiltrating cells in psoriasis patients were characterized in biopsies from both involved and uninvolved skin. Histologic examination of biopsies showed the presence of both CD4+ and CD8+ T cells and the lack of B lymphocytes. Skin biopsies were also placed in tissue culture medium supplemented with human serum, interleukin-2 (IL-2), and irradiated autologous blood lymphocytes. T lymphocytes grew from both plaques and univolved skin biopsies and consisted of a heterogeneous population of T-cell subsets. The immunophenotypic analysis of cultured cells was comparable to the histologic examination on frozen section, i.e., there was a greater number of CD4/CDw29+ cells than CD8+/CD45+ cells. Cultures were tested in the primed lymphocyte test (PLT) and cell-mediated lympholysis (CML) assays. All cultures tested demonstrated secondary proliferative but not cytolytic reactivity. The PLT results indicate that the cell cultures generated are autoreactive. This autoreactivity was found to be directed against non-human leukocyte antigens (HLA), i.e., minor HLA with some restriction to major HLA antigens.

Distribution of luteinizing hormone-releasing hormone (LHRH) in the guinea pig brain.

With the use of the unlabeled antibody enzyme technique and antiserum to luteinizing hormone-releasing hormone (LHRH), the distribution of LHRH in the adult male guinea pig was studied. Immunoreactive deposits were found in the cell bodies of hypothalamic neurons in the medial preoptic, anterior hypothalamic, suprachiasmatic, and arcuate nuclei. Ten to thirty LHRH-containing perikarya were seen per brain. Four fiber tracts positive for LHRH were described: Tract I runs on the pre-commissural side of the anterior commissure from the bed nucleus of the stria terminalis and medial preoptic area to the suprachismatic nucleus: Tract II runs on the post-commissural side to the retrochiasmatic portion of the suprachiasmatic nucleus; Tract III originates in the arcuate nucleus and the ventral portion of the ventromedial nucleus and courses medially and caudally to enter the median eminence; Tract IV travels from the mammillary bodies to the mid-brain. Three weeks after surgical isolation of the medial basal hypothalamus there was a slight decrease in the amount of LHRH in the median eminence. These data suggest that LHRH is synthesized in more than one hypothalamic nuclear group but that the majority of axons containing the hormone in the median eminence originate in the medial basal hypothalamus.

Effects of gonadectomy and treatment with gonadal steroids and luteinizing hormone secretion in hypogonadal male and female mice with preoptic area implants.

The ability of male and female hypogonadal (hpg) mice with preoptic area (POA) grafts to show negative feedback was studied. GnRH neurons within POA grafts send axons into the median eminence of the hpg host (HPG/POA), resulting in increased gonadotropin production and gonadal development in the mice that are genetically unable to produce GnRH. The present studies evaluated whether negative feedback, an aspect of normal reproductive function, is present in male and female HPG/POA mice. In normal male mice plasma LH was increased 1 or 5 months after castration and returned to baseline after testosterone propionate treatment. In contrast, no alterations in plasma LH were measured in similarly treated HPG/POA males. HPG/POA female mice were ovariectomized 6 weeks or 3 or 6 months after graft surgery and received sc 17 beta-estradiol (E2) implants 3 months later. Normal mice were studied when 6 weeks old and 8 months old (age-matched to HPG/POA mice ovariectomized 3 months after graft surgery). Further, to determine whether the mice were capable of positive feedback, 1 week after receiving E2 implants, mice in the 6-week and 3-month postgraft surgery groups were challenged with sequential administration of estradiol benzoate and progesterone. The significant increase in plasma LH after ovariectomy or decrease after E2 implant in normal female mice was not present in most HPG/POA female mice. Just 2 of the 24 HPG/POA females studied had increased plasma LH after gonadectomy, and in only 1 of these was plasma LH suppressed by E2 treatment. The ability of an individual HPG/POA mouse to show positive feedback did not predict the ability to show negative feedback, nor did the ability to show negative feedback predict positive feedback capability. Among the mice that failed to respond to ovariectomy with increased LH release were some that had elevated LH in response to steroid challenge or had spontaneously ovulated. On the other hand, neither mouse that had increased LH release after ovariectomy had shown positive feedback to a steroid challenge. Immunocytochemical evaluation revealed GnRH cells within the grafts and GnRH fiber innervation of the host's median eminence, but there was no correlation between numbers of GnRH cells or extent of innervation with the ability to show either negative or positive feedback, nor was the presence of vasoactive intestinal peptide cells within the grafts of predictive value. The failure of negative feedback in most of the HPG/POA mice tested may be due to the failure to establish as yet unidentified but essential afferents to the grafted GnRH cells and/or their axonal processes.

Effects of N-methyl-D,L-aspartic acid on luteinizing hormone secretion in normal mice and in hypogonadal mice with fetal preoptic area implants.

Many aspects of reproductive function are corrected in hypogonadal mice with preoptic area grafts (HPG/POA). Gonadotropin release and gonadal development are dependent on the presence of GnRH cells within the grafts and GnRH innervation of the median eminence. This study examined the effect of a known modulator of GnRH secretion, N-methyl-D,L-aspartic acid (NMA), in adult normal and HPG/POA male and female mice. All HPG/POA males had significant testicular development after graft surgery, and most HPG/POA females were in constant vaginal estrus and showed ovarian and uterine development; a few also demonstrated ovulatory cyclicity after pregnancies initiated by reflex ovulation. Groups of normal and HPG/POA males that were intact (INT) or castrated (CX) 7 days before testing were challenged with saline, NMA (20 mg/kg), and GnRH (100 ng/0.1 ml). Sequential blood samples from awake animals were obtained via intracardiac catheters for evaluation of plasma LH. There were significant increases in plasma LH after NMA challenge in normal INT [n = 15; 0 min, 0.26 +/- 0.02 (mean +/- SE); 10 min, 1.20 +/- 0.10 ng/ml; P less than 0.05] and normal CX (n = 13; 0 min, 0.36 +/- 0.06, 10 min, 3.25 +/- 0.27). Plasma LH secretion in response to NMA was significantly correlated (r = 0.786; P less than 0.001) with plasma LH release after the GnRH challenge in normal males. In contrast, only 3 of 17 HPG/POA (1 INT and 2 CX) showed increased circulating LH after NMA challenge, despite heightened pituitary sensitivity to GnRH. Normal and HPG/POA female mice were ovariectomized (OX) or OX and estrogen primed (OXE2) 7 days before testing. Intact cycling normal and cycling HPG/POA mice were tested in estrus (EST). There was a greater response to NMA in normal OX (n = 8; 0 min, 0.39 +/- 0.02; 10 min, 1.44 +/- 0.28) than in OXE2 (n = 13; 0 min, 0.29 +/- 0.01; 10 min, 0.52 +/- 0.07) despite similar gonadotroph sensitivity to GnRH. There was also a significant increase in plasma LH in response to NMA in HPG/POA-OX (n = 7; 0 min, 0.50 +/- 0.10; 10 min, 1.62 +/- 0.22) and HPG/POA-OXE2 (n = 12; 0 min, 0.39 +/- 0.04; 10 min, 1.31 +/- 0.26). Plasma LH levels after NMA treatment were significantly correlated with responses to GnRH in female HPG/POA (r = 0.58; P less than 0.03), but not in normal females. Neither normal-EST nor HPG/POA-EST had increased LH release after NMA challenge, perhaps due to the low gonadotroph sensitivity in this state.(ABSTRACT TRUNCATED AT 400 WORDS)

Gonadal steroids regulate the number and activational state of mast cells in the medial habenula.

While mast cells in connective tissues have long been associated with allergic reactions, it is now clear that they are also present within the central nervous system under normal physiological conditions. The mast cell population increases 10-fold in the medial habenular region of the brain within 2 h after pairing in doves. The first study explored whether this increase was due to exposure to gonadal steroids. Light microscopic immunocytochemistry indicates an increased number of brain MC following exposure to either testosterone (T) or dihydrotestosterone (DHT) in the male, or 17beta estradiol (E) in the female, but not in cholesterol-treated controls. Thus, the increased habenular MC population is produced by gonadal hormones in the absence of sexual behavior, is not sexually dimorphic, and does not require aromatization of androgen. In the next study, MC activational state was determined using electron microscopy. Cells were categorized into five states: (I) resting; (II) initiation of degranulation; (III) fully degranulated; (IV) piecemeal secretion; and (V) resynthesizing. Hormone treatment (T, DHT, or E) resulted in a significant increase in the percent of cells in activated states. MC granules contain a wide range of biologically active molecules. The release of these granule contents into the neuropil of the central nervous system is likely to have wide ranging effects at multiple levels including vascular permeability and neuronal excitability. In that steroid treatment is known to result in such effects, the present demonstration of a hormonally induced shift in MC secretory state is one avenue by which these effects are mediated.

Sexual dimorphism in the synaptic input to gonadotropin releasing hormone neurons.

GnRH neurons form the final common pathway regulating the secretion of gonadotropins from the anterior pituitary. Since the patterns of gonadotropin release display profound sexual dimorphism among mammals including the rodent, we undertook an ultrastructural analysis to determine whether these neurosecretory cells were differentially innervated between the sexes. As a further exploration of the organization of the neurocircuitry integrating GnRH neurons with the central nervous system, we also determined the degree to which GnRH cells and their processes were innervated by terminals containing either the endogenous opiate, beta-endorphin (BE) or GnRH itself. Sections from the diagonal band of Broca and the preoptic area of adult male and diestrus II female rats were immunocytochemically processed for dual localization of GnRH and BE. GnRH neurons cut through the plane of the nucleus were identified in 1 micron sections. Serial ultrathin sections were made and analyzed for 1) total synaptic input to both cell bodies and dendrites; 2) BE input; and 3) input arising from GnRH itself. We report that GnRH neuronal cell bodies in females received approximately twice the number of synapses as did those of males. The input to the GnRH dendrites, when measured as percent of plasma membrane in synaptic contact, also showed a profound sexual dimorphism with the female having a larger proportion of the dendrite in synaptic apposition. BE innervation contributed to this dimorphism at the level of both the cell body and dendrite. In contrast, the distribution and number of GnRH terminals did not differ between the sexes. In both they were confined to the dendritic arbor. We hypothesize that the capacity of the female rodent GnRH system to show neurogenic derived alterations in GnRH output not seen in the male may be due in part to these anatomical differences.