BACKGROUND The purpose of this investigation was to test the efficacy of novel antipsychotic medications in the treatment of cognitive impairment in early phase schizophrenia. METHODS Sixty-five ...patients in this multicenter double-blind study were randomly assigned to olanzapine (5-20 mg), risperidone (4-10 mg), or haloperidol (5-20 mg). Standard measures of clinical and motor syndromes were administered, as well as a comprehensive battery of tests to assess (1) motor skills, (2) attention span, (3) verbal fluency and reasoning, (4) nonverbal fluency and construction, (5) executive skills, and (6) immediate recall at baseline and after 6, 30, and 54 weeks of treatment. RESULTS The general cognitive index derived from the 6 domain scores revealed a significantly greater benefit from treatment with olanzapine relative to haloperidol and olanzapine relative to risperidone, but no significant difference was shown between risperidone and haloperidol. The improvement related to olanzapine was apparent after 6 weeks and enhanced after 30 and 54 weeks of treatment. Exploratory within-group analyses of the 6 cognitive domains after a conservative Bonferroni adjustment revealed a significant improvement with olanzapine only on the immediate recall domain, and similar analyses of the 17 individual tests revealed a significant improvement with olanzapine only on the Hooper Visual Organization Test. CONCLUSIONS These data suggest that olanzapine has some superior cognitive benefits relative to haloperidol and risperidone. A larger sample replication study is necessary to confirm and generalize the observations of this study and begin evaluation of the implications of this change to cerebral function and quality of life for people with schizophrenia.Arch Gen Psychiatry. 2000;57:249-258-->
Platelets are a recognised potent source of transforming growth factor‐β1 (TGFβ1), a cytokine known to promote wound healing and regeneration by stimulating dermal fibroblast proliferation and ...extracellular matrix deposition. Platelet lysate has been advocated as a novel personalised therapeutic to treat persistent wounds, although the precise platelet‐derived growth factors responsible for these beneficial effects have not been fully elucidated. The aim of this study was to investigate the specific role of platelet‐derived TGFβ1 in cutaneous wound healing. Using a transgenic mouse with a targeted deletion of TGFβ1 in megakaryocytes and platelets (TGFβ1fl/fl.PF4‐Cre), we show for the first time that platelet‐derived TGFβ1 contributes to epidermal and dermal thickening and cellular turnover after excisional skin wounding. In vitro studies demonstrate that human dermal fibroblasts stimulated with platelet lysate containing high levels of platelet‐derived TGFβ1 did not exhibit enhanced collagen deposition or proliferation, suggesting that platelet‐derived TGFβ1 is not a key promoter of these wound healing processes. Interestingly, human keratinocytes displayed enhanced TGFβ1‐driven proliferation in response to platelet lysate, reminiscent of our in vivo findings. In summary, our novel findings define and emphasise an important role of platelet‐derived TGFβ1 in epidermal remodelling and regeneration processes during cutaneous wound healing.
Abstract
Diffuse hemispheric gliomas, H3G34R/V-mutant (DHG-H3G34), are uniformly lethal malignancies with currently no targeted therapies available. They exclusively occur in the cerebral hemispheres ...of adolescents and young adults, and have been linked to a distinct interneuronal lineage of origin. The developmental spectrum and functional role of this interneuronal lineage in DHG-H3G34 remain incompletely understood. Here, through integrating bulk and single-cell multi-omics with genome-wide CRISPR-Cas9 screens, we resolve a putative cellular hierarchy that follows a continuum of interneuronal lineage development, ranging from a self-renewing progenitor-like cell to a more differentiated cell resembling early immature GABAergic interneurons, along with quiescent astrocyte-like and mesenchymal-like cells. We validate these single-cell states in patient DHG-H3G34 tissue sections by multiplexed immunofluorescence, and describe spatial structures that resemble nests of early migratory interneurons surrounded by progenitor cells, characteristic of human embryonal interneuron development. Intriguingly, we reveal the majority of CRISPR-Cas9 screen-derived gene dependencies are upregulated in interneuronal lineage tumor cells, specifically in less differentiated progenitor-like cells, highlighting these as a driver of DHG-H3G34. We validate the essentiality of these interneuronal lineage associated targets in patient-derived in vitro and in vivo models, and highlight CDK6 as a druggable target selectively upregulated in DHG-H3G34. Inhibition of CDK6 leads to a decrease of undifferentiated progenitor-like signatures, reduced tumor growth, and prolonged survival of patient-derived xenograft models. Encouraged by these findings, we treated a patient upon a second relapse of a DHG-H3G34 with ribociclib on a compassionate use basis, who, as of the time of submission, has shown stable disease within four cycles of ribociclib treatment after progression on PCV chemotherapy. In sum, we reveal CDK6 inhibition as a rationally informed and clinically actionable therapeutic avenue that selectively perturbs the unique interneuronal lineage in DHG-H3G34, paving the way for rapid clinical translation.
1 Department of Pharmacology and Physiology, Drexel University College of Medicine; Philadelphia;
2 Department of Surgery, Division of Urology, University of Pennsylvania School of Medicine, ...Philadelphia; and
3 Department of Obstetrics and Gynecology, Division of Urogynecology and Reconstructive Pelvic Surgery, University of Pennsylvania School of Medicine, Philadelphia, Pennsylvania/
Submitted June 18, 2008
; accepted in final form September 3, 2009
Information regarding the role of cholinergic nerves in mediating vaginal smooth muscle contraction is sparse, and in vitro studies of the effects of muscarinic agonists on vaginal smooth muscle are discrepant. The goal of this study was to determine the expression of muscarinic receptors in the vaginal wall of the rat. In addition, we sought to determine the effect of the muscarinic receptor agonist carbachol on contractility and inositol phosphate production of the proximal and distal rat vaginal muscularis. RT-PCR analysis indicated that both M 2 and M 3 receptor transcripts were expressed within the proximal and distal rat vagina. Carbachol dose-dependently (10 –7 –10 –4 M) contracted the rat vaginal muscularis with a greater maximal contractile response in the proximal vagina ( P < 0.01) compared with the distal vagina. The contractile responses of the rat vaginal muscularis to carbachol were dose dependently inhibited by the M 3 antagonist para-fluoro-hexahydrosiladefenidol, and a pK B of 7.78 and 7.95 was calculated for the proximal and distal vagina, respectively. Inositol phosphate production was significantly increased in both regions of the vagina following 20-min exposure to 50 µM carbachol with higher levels detected in the proximal vagina compared with the distal ( P < 0.05). Preliminary experiments indicated the presence of M 2 and M 3 receptors in the human vaginal muscularis as well as contraction of human vaginal muscularis to carbachol, indicating that our animal studies are relevant to human tissue. Our results provide strong evidence for the functional significance of M 3 receptor expression in the vaginal muscularis.
vagina; smooth muscle; muscarinic receptor; female sexual response
Address for reprint requests and other correspondence: S. Chacko, Basic Urology Research Laboratory, Glenolden Research Bldg., 500 S. Ridgeway Ave., Glenolden, PA 19036-2307 (e-mail: chackosk{at}mail.med.upenn.edu ).
Detrusor smooth muscle (DSM) hypertrophy induced by partial bladder outlet obstruction (PBOO) is associated with changes in the NH2-terminal myosin heavy chain isoform from predominantly SM-B to ...SM-A, alteration in the Ca2+ sensitization pathway, and the contractile characteristics from phasic to tonic in rabbits. We utilized the SM-B knockout (KO) mouse to determine whether a shift from SM-B to SM-A without PBOO is associated with changes in the signal transduction pathway mediated via PKC and CPI-17, which keeps the myosin phosphorylation (MLC20) level high by inhibiting the myosin phosphatase. DSM strips from SM-B KO mice generated more force in response to electrical field stimulation, KCl, carbachol, and phorbol 12,13-dibutyrate than that of age-matched wild-type mice. There was no difference in the ED50 for carbachol but the maximum response was greater for the SM-B KO mice. DSM from SM-B KO mice revealed increased mass and hypertrophy. The KO mice also showed an overexpression of PKC-alpha, increased levels of phospho-CPI-17, and an elevated level of IP3 and DAG upon stimulation with carbachol. Two-dimensional gel electrophoresis revealed an increased level of MLC20 phosphorylation in response to carbachol. Together, these changes may be responsible for the higher level of force generation and maintenance by the DSM from the SM-B KO bladders. In conclusion, our data show that ablation of SM-B is associated with alteration of PKC-mediated signal transduction and CPI-17-mediated Ca2+ sensitization pathway that regulate smooth muscle contraction. Interestingly, similar changes are also present in PBOO-induced DSM compensatory response in the rabbit model in which SM-B is downregulated.
To determine concentrations of 17alpha-hydroxyprogesterone (17OHP) in serum of healthy bitches during various stages of the reproductive cycle and in bitches with hyperadrenocorticism and to compare ...the dynamics of 17OHP with those of progesterone.
Prospective evaluation study.
15 healthy sexually intact bitches and 28 spayed bitches with hyperadrenocorticism.
11 healthy bitches were evaluated during estrus, nonpregnant diestrus, and anestrus (group 1); 4 other healthy bitches were evaluated during pregnancy and after ovariohysterectomy (group 2). Cycle stages were determined via physical examination, vaginal cytologic evaluation, and serum progesterone concentration. Bitches with hyperadrenocorticism were evaluated once at the time of diagnosis (group 3). Serum hormone concentrations were determined with immunoassays.
In group 1, the serum 17OHP concentration was significantly higher in diestrus (median, 1.8 ng/mL) than in estrus (median, 1.1 ng/mL) and anestrus (median, 0.2 ng/mL) and higher in estrus than in anestrus. Changes in serum progesterone concentrations accounted for 22% (estrus) or 23% (diestrus) of the variation in serum 17OHP concentrations. In group 2, 17OHP and progesterone concentrations were significantly higher during pregnancy than after ovariohysterectomy. The serum 17OHP concentration in group 3 was significantly lower (median, 0.2 ng/mL) than in group 1 in estrus and diestrus and in group 2 during pregnancy (median, 0.7 ng/mL) but was not different from 17OHP concentrations in anestrus or after ovariohysterectomy (median, 0.2 ng/mL).
Serum 17OHP concentrations in healthy bitches increased during estrus, diestrus, and pregnancy and at those times were higher than in spayed bitches with hyperadrenocorticism.
Smooth muscle hypertrophy, induced by partial bladder outlet obstruction, leads to changes in detrusor smooth muscle contractility. We analyzed lipid‐dependent signaling systems during detrusor ...remodeling following smooth muscle hypertrophy to determine whether IP3 production and arachidonic acid (AA) release from phospholipid are altered in detrusor. Rabbit bladders were partially obstructed for 2 weeks, and then smooth muscle isolated. Levels of IP3 production and AA release were compared with levels obtained with 2‐week sham‐operated rabbits. Smooth muscle hypertrophy significantly increased AA release and IP3 production in smooth muscle. Carbachol stimulated AA release from control detrusor but not from hypertrophic detrusor. AA release from smooth muscle was inhibited by AACOCF3, an inhibitor of cPLA2, which suggested that cPLA2 was required for AA release. During hypertrophy, the amount of cPLAz in smooth muscle was increased by a factor of 2. Carbachol stimulated IP3 production from control and 2 week obstructed bladders. Bladder detrusor muscle achieved optimal force production after 1–2 min of carbachol treatment, and IP3 production became optimal during the first 2 min of carbachol treatment. Neomycin inhibited force production and IP3 production in rabbit bladder detrusor. IP3 production in rabbit bladder detrusor correlates with force development and the sensitivity of both force and IP3 production to neomycin suggests that IP3 production plays a role in force development.
Source of Funding: NIH Center Grant P50 DK 52620.
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