Viral co-infection, autoimmunity, and CSF HIV antibody profiles in HIV central nervous system escape.

Antiretroviral therapy (ART) suppresses plasma and cerebrospinal fluid (CSF) HIV replication. Neurosymptomatic (NS) CSF escape is a rare exception in which CNS HIV replication occurs in the setting of neurologic impairment. The origins of NS escape are not fully understood. We performed a case-control study of asymptomatic (AS) escape and NS escape subjects with HIV-negative subjects as controls in which we investigated differential immunoreactivity to self-antigens in the CSF of NS escape by employing neuroanatomic CSF immunostaining and massively multiplexed self-antigen serology (PhIP-Seq). Additionally, we utilized pan-viral serology (VirScan) to deeply profile the CSF anti-viral antibody response and metagenomic next-generation sequencing (mNGS) for pathogen detection. We detected Epstein-Barr virus (EBV) DNA more frequently in the CSF of NS escape subjects than in AS escape subjects. Based on immunostaining and PhIP-Seq, there was evidence for increased immunoreactivity against self-antigens in NS escape CSF. Finally, VirScan revealed several immunodominant epitopes that map to the HIV envelope and gag proteins in the CSF of AS and NS escape subjects. Whether these additional inflammatory markers are byproducts of an HIV-driven process or whether they independently contribute to the neuropathogenesis of NS escape will require further study.

Efficacy and safety of a single-tablet regimen containing tenofovir disoproxil fumarate 300 mg, lamivudine 300 mg and efavirenz 400 mg as a switch strategy in virologically suppressed HIV-1-infected subjects on nonnucleoside reverse transcriptase inhibitor-containing first-line antiretroviral therapy in Pune, India.

OBJECTIVES: As per National AIDS Control Organization (NACO) estimates, there are 2.1 million people living with HIV (PWH) in India, of whom 1.2 million are on first-line antiretroviral therapy (ART). This study explored the use of a single-tablet regimen containing tenofovir disoproxil fumarate 300 mg + lamivudine 300 mg + efavirenz 400 mg (TLE400 STR) as a first-line switch strategy in PWH in Pune, India. METHODS: This retrospective cohort study was conducted in private sector ART clinics in three tertiary-level hospitals in Pune, India. PWH > 12 years of age (n = 502) who initiated first-line ART (predominantly TLE600 STR), completed ≥ 6 months of follow-up and achieved virological suppression [plasma viral load (VL) < 1000 HIV-1 RNA copies/mL] were identified and switched to TLE400 STR. The virological and immunological efficacy of TLE400 STR at 6 and 12 months of follow-up were noted. Grade 3/4 adverse events (especially efavirenz-related neuropsychiatric adverse events) leading to regimen discontinuation were also noted. RESULTS: Of 502 PWH who switched to TLE400 STR, complete virological suppression (VL < 20 copies/mL) was maintained in more than 97% of patients at follow-up. TLE400 STR was successful in maintaining CD4 counts within the range observed at the start of the regimen. Grade 3/4 adverse events leading to TLE400 STR discontinuation were seen in 11 (2.2%) patients. Virological failure (VL > 1000 copies/mL) and treatment regimen failure were seen in six (1.2%) and 49 (9.8%) subjects, respectively. CONCLUSIONS: TLE400 STR exhibits excellent efficacy and safety as a switch strategy and should be introduced in the Indian National ART Program, especially for PWH who are virologically suppressed on TLE600 STR.

Gene therapy as a potential tool for treating neuroblastoma-a focused review.

Neuroblastoma, a solid tumor caused by rapid division of undifferentiated neuroblasts, is the most common childhood malignancy affecting children aged <5 years. Several approaches and strategies developed and tested to cure neuroblastoma have met with limited success due to different reasons. Many oncogenes are deregulated during the onset and development of neuroblastoma and thus offer an opportunity to circumvent this disease if the expression of these genes is restored to normalcy. Gene therapy is a powerful tool with the potential to inhibit the deleterious effects of oncogenes by inserting corrected/normal genes into the genome. Both viral and non-viral vector-based gene therapies have been developed and adopted to deliver the target genes into neuroblastoma cells. These attempts have given hope to bringing in a new regime of treatment against neuroblastoma. A few gene-therapy-based treatment strategies have been tested in limited clinical trials yielding some positive results. This mini review is an attempt to provide an overview of the available options of gene therapy to treat neuroblastoma.

Estrous stage determination in rats by means of scanning electron microscopic images of uterine surface epithelium.

With the results presented in this paper we devised an alternative method to precisely date the rat endometrium in relation to the estrous cycle. This is done by the exclusive use of scanning electron microscopy. Owing to its short estrous cycle (4 to 6 days, depending on age), the rat is ideally suited for the examination of cyclic changes occurring in the uterine epithelium. The cycle stage of rats predated by vaginal smear cytology was verified through the measurement of hormones relevant to the estrous cycle, i.e. estradiol-17-beta (E2), progesterone (P), luteinizing hormone (LH), and follicle stimulating hormone (FSH). Based on scanning electron images surface changes could thus be correlated to the cyclic variations of blood levels of sex hormones. The appearance of pseudoglands, the most prominent aspect during the cycle, is correlated with decreasing estrogen and rising progesterone levels. Pseudoglands are formed by apoptosis and necrosis of epithelial cells, and are most numerous during estrus. They had in previous studies been classified as genuine uterine glands.

Centrally acting alpha 1-adrenoceptor agonists based on hexahydronaphth[2,3-b]-1,4-oxazines and octahydrobenzo[g]quinolines.

Centrally acting alpha 1-agonists may be of therapeutic value in dementias and other CNS disorders characterized by symptoms of noradrenergic insufficiency. Therefore, on the basis of known peripherally acting alpha 1-agonists two new groups of centrally acting alpha 1-agonists with improved lipophilicity, the hexahydronaphth[2,3-b]-1,4-oxazines type A and the octahydrobenzo[g]quinolines type B were designed. The N-methylated derivatives 14 and 33 demonstrate potent, direct agonistic activity at postjunctional alpha 1-receptors. Ring substituent alterations in compounds of type A and B change the potency of compounds on the rabbit ear artery by over 3 orders of magnitude (pD2 = 5.35-8.40). The efficacy of these compounds varies from 42 to 110%. Those alpha 1-agonists which were selective in the pithed rat increase vigilance in rats. Compound 14 was found to be a centrally acting alpha 1-agonist with good tolerability in different animal species and in healthy volunteers. Furthermore, 14 selectively stimulates the breakdown of phosphatidylinositol in rat cerebral cortex slices. In vivo, the compound reverses behavioral deficits in animals which received noradrenergic lesions following DDC or DPS4 treatment. Oxazine 14 and its close derivatives are by far more lipophilic than commonly known alpha 1-agonists. This is demonstrated in a ClogP-PROBIS plot.

5-HT1C receptor-mediated stimulation of inositol phosphate production in pig choroid plexus. A pharmacological characterization.

1) 5-HT (5-hydroxytryptamine, serotonin) induces inositol phosphate production in a pig choroid plexus preparation. This effect has been pharmacologically characterized and the data compared to those obtained from radioligand binding studies performed with [3H]mesulergine to 5-HT1C sites in pig choroid plexus membranes. 2) The rank order of potency of agonists stimulating inositol phosphate production was: alpha-methyl-5-HT greater than 1-methyl-5-HT greater than DOI greater than bufotenine = SKF 83566 = 5-HT greater than 5-MeO-DMT greater than 5-MeOT = RU 24969 greater than SCH 23390 greater than 5-CT. 8-OH-DPAT was virtually devoid of activity at 100 mumol/l. 3) The increase in inositol phosphate production induced by 5-HT and other agonists was surmountably antagonised by mesulergine, ketanserin and spiperone with pKB values of 8.7, 6.7 and 5.3, respectively. 4) The rank order of potency of antagonists was: metergoline greater than mesulergine greater than LY 53857 greater than ritanserin greater than methiothepin greater than mianserin greater than cyproheptadine greater than pirenperone greater than cinanserin greater than ketanserin greater than spiperone. The following antagonists were virtually devoid of activity at 100 mumol/l; pindolol, 21-009 and yohimbine. 5) The results obtained both with agonists and antagonists strongly support the view that 5-HT1C receptors mediate agonist induced production of inositol phosphates in pig choroid plexus.(ABSTRACT TRUNCATED AT 250 WORDS)

The effect of chronic hydergine treatment on the plasticity of synaptic junctions in the dentate gyrus of aged rats.

The number of synapses (Nv), the surface density of contact zones (Sv) as well as the average size (S) of E-PTA stained synapses in the supragranular layer of the dentate gyrus from adult (12 months), old (30 month), and Hydergine-treated old (30 months) rats were measured by using quantitative morphometric techniques. In old animals, Nv and Sv were significantly reduced, whereas S was significantly increased as compared with the values in adult rats. Hydergine (Codergocrine mesylate) treatment of old animals (3 mg/Kg/day for 4 weeks) influenced these three parameters, differentially. The Sv in aged animals receiving Hydergine, relative to that in untreated old rats, was significantly increased; the number and size of synapses in the treated old rats were significantly higher and smaller, respectively, than that in old controls. We interpret the present findings to indicate a modulating effect of Hydergine on the morphological plasticity of synaptic junctions in the dentate gyrus of aged rats.

Noradrenergic, serotonergic, and cholinergic sprouting in the hippocampus that follows partial or complete transection of the septohippocampal pathway: contributions of spared inputs.

The aminergic and cholinergic fibers innervating the hippocampus reach their target regions via the dorsal (dorsal fornix, fimbria, and cingulum) and ventral routes. The plasticity of this innervation after lesions of the dorsal pathway was investigated in the septal, medial, and temporal regions of the adult rat hippocampus. The extent and time course of sprouting was assessed by determining high-affinity uptake of [3H]noradrenaline and [3H]serotonin, and the activities of cholinergic enzymes 1 week to 6 months after partial or complete transection of the dorsal pathway. The initial decline in these terminal indices resulting from the transection of the medial or lateral half of the dorsal pathway reflected the distribution of their respective terminal sites in the three hippocampal regions. Longer survival after such lesions led to a substantial recovery of cholinergic and noradrenergic markers indicating reinnervation of the hippocampus by spared dorsal and/or by undamaged ventral afferent fibers. Whether the recovery was complete or not, the sprouting of spared dorsal fibers was maximum within a relatively shorter postlesion survival whereas that of the ventral afferents continued for a longer time. In contrast to the extent of noradrenergic and cholinergic restitution, very poor serotonergic recovery was seen in the same animal in spite of the availability of spared serotonergic afferent fibers and regardless of the duration of postlesion survival. The apparently finite capacity of spared axons for postlesion reinnervation was not increased by longer survival. Amplification of this property of central neurons by other interventions may supplement the approach of transplantation for recovery of function following injury.

Unilateral fimbria/fornix lesions attenuate behavioral symptoms induced by subsequent dorsal hippocampal lesions in rats.

Adult rats were given a unilateral fimbria-fornix (5, 10, 15 micrograms colchicine or electrolytic) lesion 30 days before a restricted bilateral dorsal hippocampal lesion to assess the behavioral effects of temporally and structurally spaced lesions. The rats were tested 30 days after the second-stage surgery in the Hebb-Williams maze learning task. All rats with lesions were impaired in comparison with sham-operated control rats. However, those which sustained electrolytic or 5 micrograms colchicine fimbria-fornix lesions before hippocampal lesions were less impaired than rats which received hippocampal lesions alone or hippocampal lesions preceded by fimbria-fornix lesions with larger doses of colchicine.

Fimbria-fornix lesion increases nerve growth factor content in adult rat septum and hippocampus.

Bilateral complete transection of the fimbria-fornix causes a significant increase in nerve growth factor (NGF) content both in the septum and hippocampus of the adult rat as measured by a sensitive immunoassay 7 days after lesioning. The finding, that elevation of NGF in septum (250%) was much more pronounced than that in the hippocampus (30-50%), cannot be explained by retrograde axonal transport. Degeneration of central cholinergic neurons might be a potential trigger for NGF production in the CNS.

Time course of the elevation of nerve growth factor (NGF) content in the hippocampus and septum following lesions of the septohippocampal pathway in rats.

Axotomy of cholinergic neurons in the medial septum-diagonal band and degeneration of their terminals in the hippocampus resulting from fornix-fimbria lesions induce elevation of NGF content in these two brain regions. Postlesion levels of cholinergic neuron-specific ChAT activity in the septum suggest that endogenously produced NGF may, at least partly, promote survival of axotomized cholinergic neurons or induce ChAT activity in undamaged cells or both. These findings thus support the proposed trophic role for NGF in central cholinergic neurons.

Homologous cholinergic efferents spared by partial fimbrial lesions contribute to the recovery of hippocampal cholinergic enzymes in adult rats.

Cholinergic fibers spared by partial lesions of the fimbrial bundle contribute to the recovery of enzyme markers for hippocampal cholinergic terminals. This recovery, most likely representing structural restoration of cholinergic terminals lost to lesion-induced degeneration, is amplified by a 'conditioning' lesion. This model of functionally relevant postlesion reconstruction, by and of structures within the CNS, is suitable to search for means to enhance homotypic collateral sprouting.

Deficits in cholinergic enzyme activities in septo-temporal regions of the senescent rat hippocampus, and in the forebrain of aged mice: effect of chronic Hydergine treatment.

Choline acetyltransferase and acetylcholinesterase activities in the septo-temporal regions of the rat hippocampus, and in the forebrain of mice were significantly decreased in senescent as compared with adult animals. A four week treatment of aged rats or mice with Hydergine resulted in the restoration of choline acetyltransferase activity.

A modulating effect of Hygergine on the synaptic plasticity of old rats.

The morphological plasticity of E-PTA stained synaptic junctions was investigated by means of quantitative morphometry in the dentate gyrus supra-granular layer of adult, old and old-Hydergine treated rats. Numerical (Nv) and surface (Sv) density as well as average size of the synapses (S) were the three parameters considered. During aging, Nv and Sv significantly decrease whereas S increases. Hydergine treatment to old rats resulted in a significant increase of Nv and Sv and a significant decrease of S. Present findings are interpreted as a modulating effect of Hydergine on the synaptic plasticity of old rats.

Recovery of enzyme markers for cholinergic terminals in septo-temporal regions of the hippocampus following selective fimbrial lesions in adult rats.

The activities of choline actyltransferase and acetylcholinesterase were determined in five consecutive septo-temporal regions of the ipsilateral and contralateral hippocampus from unlesioned controls and lesioned animals at various times following lateral, medial or complete unilateral transection of the fimbrial bundle in rats. In control animals distribution of cholinergic enzymes suggests a relatively heavier innervation of the ventral hippocampus. In lesioned animals depletion of enzyme activities in septo-temporal regions of the ipsilateral hippocampus was consonant with the known topography of cholinergic innervation of the hippocampus via the dorsal and ventral pathways. After 4 and 8 week post-lesion survival, a substantial recovery of both enzyme activities was evident following either of the lesion paradigms employed. However, the extent and the pattern of enzyme restitution depended on the type of fimbrial transection and the hippocampal region under consideration. Significant enzyme alterations were also observed in the contralateral hippocampus following all three lesion types. We interpret the lesion-induced temporal consequences in cholinergic enzymes to indicate initial degeneration and subsequent regeneration of cholinergic terminals in the hippocampus. The present findings also suggest that homologous fimbrial fibres spared by the partial lesions are responsible for the ensuing recovery. Thus, partial lesions of well-defined efferents constitute a suitable experimental paradigm to demonstrate homotypic reconstruction in the adult mammalian central nervous system.

Spontaneous recovery from motor asymmetry in adult rats with 6-hydroxydopamine-induced partial lesions of the substantia nigra.

Animals with 6-hydroxydopamine-induced partial unilateral lesions of the substantia nigra exhibit spontaneous recovery from motor asymmetry, a transitory increase in dopamine turnover and an increase in tyrosine hydroxylase activity in the denervated striatum. The recovery of function in these animals seems to be due to the compensatory increase in dopamine metabolism as well as due to the time-dependent increase in tyrosine hydroxylase resulting from either enzyme activation or following reinnervation of the denervated striatum by nigral efferents spared by the partial lesions.

Classification of drugs according to receptor binding profiles.

The affinity for eight different neurotransmitter receptors of about forty drugs, used for the treatment of various central nervous system disorders, was determined following in vitro receptor binding assays. Our findings indicate that, in spite of widely varying chemical structures and often poorly understood mechanisms of action, the similarities in the "affinity profiles" permit a clinically meaningful classification of these drugs. Such an approach would thus be useful in the assessment of newly synthesized compounds at an early stage of drug development.

Chronic intraventricular injections of nerve growth factor elevate hippocampal choline acetyltransferase activity in adult rats with partial septo-hippocampal lesions.

Nerve growth factor (NGF) was injected intraventricularly during 4 weeks into adult rats with unilateral partial lesions of the cholinergic septo-hippocampal pathway. On the lesioned side, NGF treatment elevated choline acetyltransferase (ChAT) activity up to 60% above the activity measured on the lesioned side of cytochrome c-treated controls. On the unlesioned side, NGF treatment increased ChAT activity only to an insignificant degree. ChAT activity in the septum of NGF-treated animals was increased by 60% as compared to controls. The NGF-induced increases on the lesioned side and in the septum were not accompanied by elevations in acetylcholinesterase (AChE) activity. Furthermore, histochemical analysis revealed no difference in AChE staining pattern or intensity between NGF-treated and control animals. The lack of effect on AChE strongly suggests that the increases in ChAT activity in hippocampus and septum are due to an elevation of ChAT activity within cholinergic neurons surviving the lesion rather than to a promotion of sprouting of cholinergic fibers.

Recovery of choline acetyltransferase and acetylcholinesterase activities in the ipsilateral hippocampus following unilateral, partial transection of the fimbria in rats.

An initial decrement followed by a significant recovery of choline acetyltransferase and acetylcholinesterase activities in the ipsilateral hippocampus was observed in rats with unilateral, partial transection of the fimbria. Regeneration of hippocampal cholinergic terminals could account for the parallel return towards normal of both the enzyme activities. We suggest that the still intact fimbrial fibres, homologous to those severed, contribute to the observed recovery.