Concurrent advanced HIV disease and viral load suppression in a high-burden setting: Findings from the 2015-6 ZIMPHIA survey.

BACKGROUND: As Zimbabwe approaches epidemic control of HIV, programs now prioritize viral load over CD4 monitoring, making it difficult to identify persons living with HIV (PLHIV) suffering from advanced disease (AD). We present an analysis of cross-sectional ZIMPHIA data, highlighting PLHIV with AD and concurrent viral load suppression (VLS). METHODS: ZIMPHIA collected blood specimens for HIV testing from 22,501 consenting adults (ages 15 years and older); 3,466 PLHIV had CD4 and VL results. Household HIV testing used the national serial algorithm, and those testing positive then received point-of-care CD4 enumeration with subsequent VL testing. We used logistic regression analysis to explore factors associated with concurrent AD and VLS (<1000 copies/mL). All analyses were weighted to account for complex survey design. RESULTS: Of the 3,466 PLHIV in the survey with CD4 and VL results, 17% were found to have AD (CD4<200cells/mm3). Of all AD patients, 30% had VLS. Concurrent AD and VLS was associated with male sex (aOR 2.45 95%CI 1.61-3.72), older age (35-49 years [aOR 2.46 95%CI 1.03-5.91] and 50+ years [aOR 4.82 95%CI 2.02-11.46] vs 15-24 years), and ART duration (<6 months [aOR 0.46 95%CI 0.29-0.76] and 6-24 months [aOR 2.07 95%CI 1.35-3.17] vs more than 2 years). The relationship between sex and AD is driven by age with significant associations among men aged 25-34, (aOR 3.37 95%CI 1.35-8.41), 35-49 (aOR 5.13 95%CI 2.16-12.18), and 50+ (aOR 12.56 95%CI 4.82-32.72) versus men aged 15-24. CONCLUSIONS: The percentage of PLHIV with AD and VLS illustrates the conundrum of decreased support for CD4 monitoring, as these patients may not receive appropriate clinical services for advanced HIV disease. In high-prevalence settings such as Zimbabwe, CD4 monitoring support warrants further consideration to differentiate care appropriately for the most vulnerable PLHIV. Males may need to be prioritized, given their over-representation in this sub-population.

Calretinin: a gene for a novel calcium-binding protein expressed principally in neurons.

A novel gene of the calmodulin superfamily, encoding a 29-kD neuronal protein here named "calretinin," has been isolated as a cDNA clone from chick retina. The encoded sequence includes four putative calcium-binding sites and a fusion protein binds calcium. The most similar protein known is the 28-kD intestinal calcium-binding protein, calbindin (58% homology). Both genes date from before the divergence of chicks from mammals. The distribution of calretinin and calbindin mRNAs in chick tissues has been mapped using RNA gel blots and in situ hybridization. RNAs from both genes are abundant in the retina and in many areas of the brain, but calretinin RNA is absent from intestine and other nonneural tissues. Calretinin and calbindin are expressed in different sets of neurons throughout the brain. Calretinin RNA is particularly abundant in auditory neurons with precisely timed discharges.

How were introns inserted into nuclear genes?

There is now abundant evidence that many introns have been inserted into nuclear genes after the divergence of multigene families, sometimes in a semi-regular pattern with respect to pre-existing domains. This note examines ways in which these insertions might have occurred using known molecular mechanisms.

RGS4 causes increased mortality and reduced cardiac hypertrophy in response to pressure overload.

RGS family members are GTPase-activating proteins (GAPs) for heterotrimeric G proteins. There is evidence that altered RGS gene expression may contribute to the pathogenesis of cardiac hypertrophy and failure. We investigated the ability of RGS4 to modulate cardiac physiology using a transgenic mouse model. Overexpression of RGS4 in postnatal ventricular tissue did not affect cardiac morphology or basal cardiac function, but markedly compromised the ability of the heart to adapt to transverse aortic constriction (TAC). In contrast to wild-type mice, the transgenic animals developed significantly reduced ventricular hypertrophy in response to pressure overload and also did not exhibit induction of the cardiac "fetal" gene program. TAC of the transgenic mice caused a rapid decompensation in most animals characterized by left ventricular dilatation, depressed systolic function, and increased postoperative mortality when compared with nontransgenic littermates. These results implicate RGS proteins as a crucial component of the signaling pathway involved in both the cardiac response to acute ventricular pressure overload and the cardiac hypertrophic program.

Calcium-binding proteins in the nervous system.

Among the many calcium-binding proteins in the nervous system, parvalbumin, calbindin-D28K and calretinin are particularly striking in their abundance and in the specificity of their distribution. They can be found in different subsets of neurons in many brain regions. Although it is not yet known whether they play a 'triggering' role like calmodulin, or merely act as buffers to modulate cytosolic calcium transients, they are valuable markers of neuronal subpopulations for anatomical and developmental studies.

Neurocan is upregulated in injured brain and in cytokine-treated astrocytes.

Injury to the CNS results in the formation of the glial scar, a primarily astrocytic structure that represents an obstacle to regrowing axons. Chondroitin sulfate proteoglycans (CSPG) are greatly upregulated in the glial scar, and a large body of evidence suggests that these molecules are inhibitory to axon regeneration. We show that the CSPG neurocan, which is expressed in the CNS, exerts a repulsive effect on growing cerebellar axons. Expression of neurocan was examined in the normal and damaged CNS. Frozen sections labeled with anti-neurocan monoclonal antibodies 7 d after a unilateral knife lesion to the cerebral cortex revealed an upregulation of neurocan around the lesion. Western blot analysis of extracts prepared from injured and uninjured tissue also revealed substantially more neurocan in the injured CNS. Western blot analysis revealed neurocan and the processed forms neurocan-C and neurocan-130 to be present in the conditioned medium of highly purified rat astrocytes. The amount detected was increased by transforming growth factor beta and to a greater extent by epidermal growth factor and was decreased by platelet-derived growth factor and, to a lesser extent, by interferon gamma. O-2A lineage cells were also capable of synthesizing and processing neurocan. Immunocytochemistry revealed neurocan to be deposited on the substrate around and under astrocytes but not on the cells. Astrocytes therefore lack the means to retain neurocan at the cell surface. These findings raise the possibility that neurocan interferes with axonal regeneration after CNS injury.

Comparing astrocytic cell lines that are inhibitory or permissive for axon growth: the major axon-inhibitory proteoglycan is NG2.

Astrocytes, oligodendrocytes, and oligodendrocyte/type 2 astrocyte progenitors (O2A cells) can all produce molecules that inhibit axon regeneration. We have shown previously that inhibition of axon growth by astrocytes involves proteoglycans. To identify inhibitory mechanisms, we created astrocyte cell lines that are permissive or nonpermissive and showed that nonpermissive cells produce inhibitory chondroitin sulfate proteoglycans (CS-PGs). We have now tested these cell lines for the production and inhibitory function of known large CS-PGs. The most inhibitory line, Neu7, produces three CS-PGs in much greater amounts than the other cell lines: NG2, versican, and the CS-56 antigen. The contribution of NG2 to inhibition by the cells was tested using a function-blocking antibody. This allowed increased growth of dorsal root ganglion (DRG) axons over Neu7 cells and matrix and greatly increased the proportion of cortical axons able to cross from permissive A7 cells onto inhibitory Neu7 cells; CS-56 antibody had a similar effect. Inhibitory fractions of conditioned medium contained NG2 coupled to CS glycosaminoglycan chains, whereas noninhibitory fractions contained NG2 without CS chains. Enzyme preparations that facilitated axon growth in Neu7 cultures were shown to either degrade the NG2 core protein or remove CS chains. Versican is present as patches on Neu7 monolayers, but DRG axons do not avoid these patches. Therefore, NG2 appears to be the major axon-inhibitory factor made by Neu7 astrocytes. In the CNS, NG2 is expressed by O2A cells, which react rapidly after injury to produce a dense NG2-rich network, and by some reactive astrocytes. Our results suggest that NG2 may be a major obstacle to axon regeneration.

Long interspersed sequences in mammalian DNA. Properties of newly identified specimens.

The genomes of primates and of rodents contain numerous long interspersed sequences or LINEs, which are mutually homologous and show characteristics of inserted reverse transcripts or retroposons. Here I report the identification of five new specimens in published DNA sequences, including the first two examples from the rat. These specimens demonstrate the generality of certain sequence arrangements seen in LINEs, viz.: 5' truncation; internal inversion with deletion; clustering with other retroposons; and evolutionary divergence at the 3' end. The 3' segments show a patchwork pattern of homology suggestive of frequent sequence exchanges between multiple subfamilies.

Eph receptors and ephrins demarcate cerebellar lobules before and during their formation.

The formation of the ten cerebellar lobules is an unsolved problem in brain development. We report a screen for the four subfamilies of Eph receptors and their ligands (ephrins) in developing mouse cerebellum, using soluble receptor-immunoglobulin and ligand-immunoglobulin fusion proteins, and antibodies against EphA and ephrin-B proteins. Our results identify Eph receptors and ephrins as the first molecules known to demarcate individual lobules during development. Staining for ephrin-A ligands is in lobule VIII as it forms, across the whole width of the cerebellum. Staining for three EphA receptors approximately coincides with presumptive lobules VI and/or VII before and just after birth, whereas a fourth EphA receptor (EphA4, which binds ligands of both subfamilies) has more widespread expression. Staining for EphB receptors is in lobules VII, VIII, and IX. Staining for ephrin-B ligands is much weaker, becomes detectable only after birth, and does not appear to be lobule-specific. Staining for all subfamilies spreads to at least some adjacent lobules as maturation proceeds. The lobule-specific patterns appear before the lobules form, and initially extend across the width of the cerebellum, in spite of the lesser conservation of the lateral extensions of the lobules. These expression patterns define previously unknown developmental units and suggest that Eph family proteins may contribute to cerebellar morphogenesis.

Aspects of depression associated with borderline personality disorder.

OBJECTIVE: Shared symptoms between borderline personality disorder and depression have resulted in inherent difficulties in evaluating the relationship between these disorders. Some theorists have argued that depression in patients with borderline personality disorder is qualitatively distinct from depression in nonborderline patients. The purpose of this study was to empirically identify aspects of depression most associated with borderline personality disorder. METHOD: Through interview and self-report measures, the authors studied depression in 50 inpatients, 21 of whom had borderline personality disorder. RESULTS: The aspects of depression most associated with borderline personality disorder were self-condemnation, emptiness, abandonment fears, self-destructiveness, and hopelessness; boredom and somatic complaints exhibited no association. CONCLUSIONS: Depression associated with borderline pathology appears to be in some respects unique, as well as distinct from nonborderline depression. The study's implications delineate the importance of considering the phenomenological aspects of depression in borderline personality disorder.

The role of introns in evolution.

What are the roles of 'classical' introns in the evolution of nuclear genes, and what was the origin of these introns? Exon shuffling has been important in the evolution of cell surface and extracellular proteins, but the evidence for it in respect of intracellular proteins is weak. Intron distributions imply that some introns have been removed while others have been inserted in the course of evolution: ancestral patterns of introns may thus have been obscured. Recent evidence on the self-splicing and reverse-splicing abilities of Group II introns supports the hypothesis that these could have been the ancestors of classical introns.

Two calcium-binding proteins mark many chick sensory neurons.

The first immunohistochemical results with a new neuronal calcium-binding protein, calretinin, are presented. Calretinin is related to the 28,000 mol. wt calcium-binding protein, calbindin, and a survey of the chick brain by in situ hybridization has identified the brain nuclei that expressed the genes for the two proteins [Rogers J.H., J. Cell Biol. 105, 1343 (1987)]. Now, antisera have been raised against calretinin fusion proteins in order to visualize individual neurons. The antisera have been used in an immunohistochemical survey of calretinin and calbindin in the chick sensory nuclei and ganglia, where these two proteins are found to be particularly prevalent. In the central nervous system, they are seen in many secondary sensory neurons and local circuit neurons, the two proteins being almost always in separate cells. However, in ganglion cells of the spinal nerves, inner ear, and retina, they are often expressed together. Their distribution in the brain is generally different from that of a third calcium-binding protein, parvalbumin. These proteins may modulate many important calcium-dependent processes in neurons, and probably have multiple functions.

Immunoreactivity for calretinin and other calcium-binding proteins in cerebellum.

Two calcium-binding proteins, calbindin and parvalbumin, have been reported to be abundant in Purkinje cells and other cell types in the cerebellum. Immunoreactivity for a related protein, calretinin, is now reported in cerebellum of chick and rat. In the chick, antibodies against calretinin stain mossy fibres throughout, and climbing fibres in a distinct group of folia. They also stain several cell types in the molecular layer. As there is no detectable calretinin mRNA in the cerebellar cortex, this cellular staining may be due to cross-reaction with an unknown antigen. In the rat, antibodies against calretinin stain the Lugaro cells, and some granule cells in lobe X; they also give weak staining of all the granule cells in the other lobes. Thus almost all the neuronal cell types in the cerebellum show immunoreactivity for at least one of the calcium-binding proteins in one or both species.

Calretinin in rat brain: an immunohistochemical study.

Calretinin is a calcium-binding protein related to calbindin-D28k; both are present in different though overlapping sets of neurons in brains of birds and mammals. We describe in detail the pattern of calretinin immunoreactivity in the rat brain. As in chick brain, calretinin immunoreactivity is abundant in various sensory pathways (particularly certain cells and fibres of the cochlear nuclei and olfactory bulb), in the heterogeneous parts of the brainstem and in parts of the hypothalamus. Many primary sensory fibres are strongly positive. Major groups of calretinin-positive neurons also include the thalamic reticular nucleus, triangular septal nucleus, lateral mammillary nucleus and substantia nigra pars compacta. Many other calretinin-positive cells are recognizable as local inhibitory neurons. Calretinin is absent from all but a few cells in the cerebral cortex, and is never found in motor neurons. There are also some distinctive positive structures whose identity is uncertain, notably irregular "shells" of cells and fibres around the thalamus and in the amygdala and an unnamed cell type in the vestibulocerebellum.

Calretinin and calbindin-D28k in rat brain: patterns of partial co-localization.

Calretinin and calbindin-D28k are homologous calcium-binding proteins, each present in a variety of neurons in the brain. Their distributions in the rat brain have been compared at the cellular level to determine whether they tend to occur in the same or in different cells, and to determine whether calbindin-positive cells show any common features once crossreaction with calretinin has been eliminated. The results show great heterogeneity. Most cells which contain one of the proteins do not contain the other, but many cells do contain both; even in the ventral cochlear nucleus, where there is abundant calretinin and most calbindin-like immunoreactivity is due to crossreaction, a few cells contain both proteins. In the substantia nigra and ventral tegmental area, many cells are double-positive but some only contain one or the other protein. Only the triangular septal nucleus is uniformly positive for both proteins. Cells which look like local-circuit neurons in many forebrain areas (cortex, hippocampus, olfactory bulb, anterior olfactory nucleus) are exclusively positive for either calretinin or calbindin, in spite of their similar morphology. In the more heterogeneous parts of the brain (including hypothalamus central gray and substantia gelatinosa), there are mixtures of calretinin-positive, calbindin-positive, and double-positive cells. In comparison with previous data on the chick, some aspects of the distributions are conserved, but double-positive cells are more frequent in the rat. The degree of heterogeneity observed, even within comparatively well-defined neuronal populations, makes it difficult to infer in what neuronal properties these proteins could be involved.

Carbonic anhydrase-II messenger RNA in neurons and glia of chick brain: mapping by in situ hybridization.

The enzyme carbonic anhydrase is widespread in brain tissue. In rodent brains it has been reported to be exclusively in oligodendroglia but there has been some debate about the generality of this finding. To investigate the cellular distribution of carbonic anhydrase by an independent technique, we have examined the chick brain by in situ hybridization to detect mRNA from the carbonic anhydrase-II gene, using as controls the actin and vimentin genes. The most intense carbonic anhydrase-II hybridization is to the choroid plexus, to the Bergmann glia of the cerebellum, and to the Müller cells in the retina. Elsewhere, some brain regions are negative while others show many individual strongly positive cells; carbonic anhydrase-II mRNA is particularly abundant in some parts of the hyperstriatum, tectum and thalamus. Some of the larger labelled cells are identifiable as neurons. By histochemistry, we confirm the presence of the carbonic anhydrase enzyme in choroid plexus and Bergmann glia, but the enzyme is also present in blood vessel walls where there is no carbonic anhydrase-II mRNA; this may be a different isozyme. During embryogenesis, carbonic anhydrase-II mRNA appears in the retina as early as two days of incubation, but does not appear in the brain until much later.

Ebstein's anomaly of the left atrioventricular valve with congenital corrected transposition of the great arteries. Diagnosis by intracavitary electrocardiography.

We present the findings in a 13-month-old infant with angiographically confirmed congenital corrected transposition of the great arteries and insufficiency of the left atrioventricular valve. Simultaneous intracavitary electrocardiographic and pressure recordings across the left atrioventricular valve were similar to those obtained in Ebsteins' anomaly and suggested Ebstein's disease of the left atrioventricular valve. To our knowledge, this is the first reported case with intracavitary electrocardiograms in a patient with congenital corrected transposition of the great arteries with Ebstein's malformation of the left atrioventricular valve. The usefulness of the simultaneous recording of the intracavitary ECG and pressure in the diagnosis by catheterization of Ebstein's anomaly of the left atrioventricular valve in patients with congenital corrected transposition of the great arteries is emphasized.

Segregation of the receptor EphA7 from its tyrosine kinase-negative isoform on neurons in adult mouse brain.

The EphA7 gene encodes not only a typical receptor tyrosine kinase (TK+) but also an isoform lacking the tyrosine kinase domain (TK-). We have made antibodies to localise EphA7 TK+ and TK- isoforms in mouse brain. The TK- isoform was not detectable prenatally, despite reported expression of the TK- mRNA in the embryo. However, both TK+ and TK- isoforms showed striking distributions in adult brain. TK+ receptor immunoreactivity was strong in neuropil throughout most of the telencephalon, probably on fine arborisations from neurons which expressed EphA7 during development (in cerebral cortex, hippocampus, and striatum). In contrast, TK- receptor immunoreactivity was conspicuous on cell bodies and proximal dendrites of a limited number of neuronal types, some of which carried EphA7 TK+ receptor on their axons. This suggests that the TK- receptor, acting as a dominant negative antagonist, may ensure that the TK+ receptor only responds to signals encountered by the growing extremities of axons or dendrites.

Distribution of the receptor EphA7 and its ligands in development of the mouse nervous system.

EphA7 is a receptor tyrosine kinase of the Eph family. We have mapped EphA7 immunoreactivity and ligand binding in mouse embryo heads and developing brain. Immunoreactivity for the full-length receptor is found in all the cell populations that express EphA7 mRNA. In particular, it is located on growing axons from EphA7-expressing neurons, both in the trigeminal nerve and in developing brain. In many cases it persists in terminal fields in adult brain. Ligand is detected in a largely complementary distribution in embryos, but is surprisingly weak or undetectable in the target regions of many EphA7-positive axons postnatally.

Matrix metalloproteases and their inhibitors are produced by overlapping populations of activated astrocytes.

Matrix metalloproteases (MMPs) and tissue inhibitors of metalloproteases (TIMPs) are involved in many cell migration phenomena and produced by many cell types, including neurons and glia. To assess their possible roles in brain injury and regeneration, we investigate their production by glial cells, after brain injury and in tissue culture, and we investigate whether they are capable of digesting known axon-inhibitory proteoglycans. To determine the action of MMPs, we incubated astrocyte conditioned medium with activated MMPs, then did western blots for several chondroitin sulphate proteoglycans. MMP-3 digested all five proteoglycans tested, whereas MMP-2 digested only two and MMP-9 none. To determine whether MMPs or TIMPs are produced by astrocytes in vitro, we tested both primary cultures and astrocyte cell lines by western blotting, and compared them with Schwann cells. All cultures produced at least some MMPs and TIMPs, with no obvious correlation with the ability of axons to grow on those cells. Both MMP-9 and TIMP-3 were regulated by various cytokines. To determine which cells produce MMPs and TIMPs after brain injury, we made lesions of adult rat cortex, and did immunohistochemistry. MMP-2 was seen to be induced in activated astrocytes through the whole thickness of the cortex but not deeper, but MMP-3 was not seen in the injured brain. TIMP-2 and TIMP-3 immunoreactivities were induced in activated astrocytes in deep cortex and the underlying white matter. In situ hybridisation confirmed induction of TIMP-2 in glia as well as neurons, but showed no expression of TIMP-4. These results show that both MMPs and TIMPs are produced by some astrocytes, but TIMP production is particularly strong, especially in deep cortex and white matter which is more inhibitory for axon regeneration. Conversely the MMPs produced may not be adequate to promote migration of cells and axons within the glial scar.