Miracle-dz/UniFAR-FTUs huggingface dataset:
Altered gap junctional communication, intercellular signaling, and growth in cultured astrocytes deficient in connexin43

Astrocytes are characterized by extensive intercellular communication mediated primarily by gap junction channels composed of connexin43. To examine this junctional protein in astrocytic functions, astrocytes were cultured from embryonic mice with a null mutation in the connexin43 gene (Reaume et al.: Science 267:1831–1834, 1995). Using anti‐Cx43 antibodies, immunoblotting and immunostaining indicated that homozygous null astrocytes were devoid of Cx43. They are also deficient in intercellular dye transfer. Astrocytes cultured from heterozygous embryos express significantly lower Cx43 compared to wild type, and their dye coupling is reduced. Markers of glial differentiation, such as glial fibrillary acidic protein and S100, appeared similar in all genotypes. Measurement of intercellular calcium concentration following mechanical stimulation of confluent astrocytes revealed that the number of cells affected by a rise in intracellular calcium was reduced in homozygous cultures compared to wild type. In fact, the calcium response in homozygous astrocytes was similar to that observed in wild‐type astrocytes in the presence of a gap junction blocker. The growth rate of astrocytes lacking Cx43 was reduced compared to wild‐type astrocytes. These results suggest that gap junctional intercellular communication mediated by Cx43 is not critical for astrocyte differentiation but is likely involved in the regulation of intercellular calcium signaling and cell growth. J. Neurosci. Res. 49:528–540, 1997. © 1997 Wiley‐Liss, Inc.
Fatal Caprine arthritis encephalitis virus–like infection in 4 Rocky Mountain goats (Oreamnos americanus)

Over a 3.5-year period, 4 Rocky Mountain goats (Oreamnos americanus), housed at a single facility, developed clinical disease attributed to infection by Caprine arthritis encephalitis virus (CAEV). Ages ranged from 1 to 10 years. Three of the goats, a 1-year-old female, a 2-year-old male, and a 5-year-old male, had been fed raw domestic goat milk from a single source that was later found to have CAEV on the premises. The fourth animal, a 10-year-old male, had not ingested domestic goat milk but had been housed with the other 3 Rocky Mountain goats. All 4 animals had clinical signs of pneumonia prior to death. At necropsy, findings in lungs included marked diffuse interstitial pneumonia characterized histologically by severe lymphoplasmacytic infiltrates with massive alveolar proteinosis, interstitial fibrosis, and type II pneumocyte hyperplasia. One animal also developed left-sided hemiparesis, and locally extensive lymphoplasmacytic myeloencephalitis was present in the cranial cervical spinal cord. Two animals had joint effusions, as well as severe lymphoplasmacytic and ulcerative synovitis. Immunohistochemical staining of fixed sections of lung tissue from all 4 goats, as well as spinal cord in 1 affected animal, and synovium from 2 affected animals were positive for CAEV antigen. Serology testing for anti-CAEV antibodies was positive in the 2 goats tested. The cases suggest that Rocky Mountain goats are susceptible to naturally occurring CAEV infection, that CAEV from domestic goats can be transmitted to this species through infected milk and by horizontal transmission, and that viral infection can result in clinically severe multisystemic disease.
Type 1 collagen marker of bone turnover, insulin-like growth factor, and leptin in dichorionic twins with discordant birth weight.

OBJECTIVE We investigated the relationship between IGF-I-IGF binding protein (IGFBP)-1 and leptin levels with type 1 collagen markers of bone turnover in dichorionic twins with or without discordant birth weight of 20% or greater. METHODS Maternal and cord bloods were collected from gestational age-matched dichorionic twins with (n = 16) or without (n = 16) discordant birth weight. The samples were assayed for cross-linked carboxyl terminal telopeptide (ICTP, a marker of bone resorption) and propeptide (PICP, a marker of bone formation) of type I collagen, leptin, IGF-I, and IGFBP-1 by RIA. RESULTS The intrauterine growth-restricted (IUGR) twins of the discordant group had higher fetal ICTP (P < 0.001) and IGFBP-1 (P < 0.001) levels, whereas PICP (P < 0.001), IGF-I (P < 0.001), and leptin (P < 0.001) were lower than the cotwins with normal weight (AGA). In contrast, concentrations of IGF-I, IGFBP-1, ICTP, PICP, and leptin were comparable between concordant twin pairs. Leptin levels were positively correlated with PICP (r = 0.61; P < 0.001) and negatively with ICTP (r = -0.57; P < 0.001) in concordant and AGA twins but not in IUGR twins. In IUGR twins, IGF-I had positive association with PICP (r = 0.76; P < 0.001) and negative association with ICTP (r= -0.76; P < 0.001), whereas IGFBP-1 was negatively correlated with PICP levels (r = -0.65; P < 0.01). No such association was found in concordant and AGA twins. CONCLUSION These data suggest that IUGR twins had high bone turnover, which is independent of maternal factors and perhaps may be due to altered IGF axis.
Change of Dynamic Gastrocnemius and Soleus Muscle Length After Block of Spastic Calf Muscle in Cerebral Palsy

Bang MS, Chung SG, Kim SB, Kim SJ: Change of dynamic gastrocnemius and soleus muscle length after block of spastic calf muscle in cerebral palsy. Am J Phys Med Rehabil 2002;81:760–764. Objective The purpose of this study was to evaluate the efficacy of gastrocnemius and soleus muscle length measured by gait analysis for characterizing the spastic calf muscles in ambulant cerebral palsy. Methods A total of 20 limbs in the 16 children with diplegic or hemiplegic cerebral palsy showing spasticity of the calf muscle were analyzed by three-dimensional gait analysis before and after botulinum toxin injection. Results The mean maximal ankle dorsiflexion angle, the minimal knee flexion angle in stance, and the dynamic gastrocnemius and soleus muscle lengths increased after the block. Paradoxically the dynamic gastrocnemius muscle lengths decreased in the eight patients after the block, all of whom had a common characteristic of the genu recurvatum. In contrast, dynamic soleus length, knee angle, and ankle angle improved after the injection. Conclusion Gastrocnemius length and soleus length are useful variables to evaluate the antispastic effect of botulinum toxin in patients with cerebral palsy. When the genu recurvatum is present, the soleus muscle length is more informative than gastrocnemius length.
[Effect of growth differentiation factor 7 on tenogenic differentiation of bone marrow mesenchymal stem cells of rat in vitro].

OBJECTIVE To investigate the effect of growth differentiation factor 7 (GDF-7) on the tenogenic differentiation of bone marrow mesenchymal stem cells (BMSCs) in vitro, to provide evidence for improving the efficacy of BMSCs on tendon repair. METHODS BMSCs were isolated from bone marrow tissue of green fluorescent protein rats by density gradient centrifugation method. Chondrogenic, osteogenic, and adipogenic differentiation assays were used to demonstrate the multi-differentiation potential of the BMSCs. BMSCs at passage 3 were cultured and divided into 4 groups according to different concentrations of GDF-7 (0, 12.5, 25.0, and 50.0 ng/mL): group A, B, C, and D, respectively. After cultured for 2 weeks in vitro, the mRNA expressions of scleraxis, tenomodulin, tenascin C, and collagen type I were detected by real-time fluorescent quantitative PCR method, the protein expressions of tenomodulin, tenascin C, and collagen type I by immunocytochemistry staining in 4 groups, and the protein expressions of tenomodulin by Western blot in groups A and C. RESULTS BMSCs had osteogenic, chondrogenic, and adipogenic differentiation potentials. The mRNA expressions of tenomodulin in groups B, C, and D were 2.85, 3.41, and 3.07 times higher than that in group A, respectively; the mRNA expressions of scleraxis in groups B, C, and D were 2.13, 1.50, and 2.56 times higher than that in group A, respectively; and the mRNA expressions of tenascin C in groups B, C, and D were 2.45, 2.86, and 1.88 times higher than that in group A, respectively. There were significant differences between groups B, C, D and group A (P < 0.05), while there was no significant difference among groups B, C, and D (P > 0.05). The mRNA expressions of collagen type I in groups B and C were 1.92 and 2.45 times higher than that in group A, showing significant differences between groups B, C and group A (P < 0.05), but no significant difference between groups A and D (P > 0.05). Immunocytochemistry staining showed that the protein expressions of tenomodulin, tenascin C, and collagen type I were detected in groups B, C, and D but not in group A. The results were further confirmed by Western blot results which showed higher protein expression of tenomodulin in group C than in group A. CONCLUSION GDF-7 can be used to promote tenogenic differentiation of rat BMSCs in vitro.
Clozapine-Associated Neuroleptic Malignant Syndrome: Two New Cases and a Review of the Literature

BACKGROUND: Clozapine has recently been found to be associated with neuroleptic malignant syndrome (NMS). Our objective is to determine if clozapine causes NMS, if the presentation of clozapine-induced NMS differs from that of traditional agents, and which set of diagnostic criteria will most readily allow diagnosis of NMS associated with clozapine. METHODS: Two new cases of clozapine-associated NMS are presented, along with previously reported cases from the literature, identified by using a MEDLINE search (1966-August 1998). From all cases, concomitant medications and washout periods were examined (if available) to assess clozapine as the likely cause of NMS. Characteristics of clozapine and traditional antipsychotic-induced NMS were compared. Different diagnostic criteria for NMS were applied to the cases to determine which were more likely to diagnose the syndrome. RESULTS: Clozapine was deemed a highly probable cause of NMS in 14 cases, a medium probability cause in five cases, and a low probability cause in eight cases. The most commonly reported clinical features were tachycardia, mental status changes, and diaphoresis. Fever, rigidity, and elevated creatine kinase were less prominent than in NMS associated with classical neuroleptics. CONCLUSIONS: Clozapine appears to cause NMS, although the presentation may be different than that of traditional antipsychotics. Levenson's original and Add-Onizio's modified criteria were more likely to diagnose NMS than were other criteria. Clozapine-associated NMS may present with fewer clinical features. Limitations are the lack of detailed information provided by many of the case reports and the use of “modified” diagnostic criteria for retrospective diagnosis.
Bcl-2 and the Outer Mitochondrial Membrane in the Inactivation of Cytochrome c during Fas-mediated Apoptosis*

Fas-driven apoptosis in Jurkat cells results in the inactivation of cytochrome c with cessation of oxygen consumption. Overexpression of Bcl-2 was found to protect against acidification and apoptosis mediated by Fas ligation in these cells. Bcl-2 is present in the outer mitochondrial membrane, but the molecular mechanism by which it protects cells is unknown. Because Bcl-2 projects into the mitochondrial intermembrane space and cytochrome cis located in the intermembrane space, we considered the possibility that Bcl-2 might protect cytochrome c from inactivation during Fas-mediated apoptosis. The present study shows that 1) in Jurkat cells, cytochrome c inactivation during Fas-driven apoptosis requires the permeabilization of the outer mitochondrial membrane; and 2) the post-mitochondrial fraction from CEM cells that overexpress Bcl-2 both prevents and reverses cytochrome cinactivation.
Association between interleukin-10 polymorphisms and Alzheimer's disease: a systematic review and meta-analysis.

UNLABELLED It has been hypothesized that polymorphisms of interleukin (IL)-10 genes affect the risk of developing late onset Alzheimer's disease (AD). However, results of different studies are often inconsistent. Our aim was to investigate by meta-analysis the association of the common polymorphisms comprehensively defining the genetic variability of the IL-10 gene with AD risk. Fifteen studies investigating the association between IL-10 polymorphisms (-1082, -819, -592) and AD were found and analyzed. The model-free approach was applied to meta-analyze these case-control genetic association studies. Available data suggested an association between -1082 polymorphism and AD risk with a marginal statistical significance (GG versus AG/AA pooled odds ratio [OR]: 0.82, 95% confidence interval CI: 0.65-1.02) and evidence of a moderate degree of between-study heterogeneity (χ2 = 27.13, d.f. = 13, p = 0.01, I2 = 52%). For the -819 and -592 polymorphisms, we did not find an association with AD, but significant between-study heterogeneity made genotype data pooling unacceptable. Analysis by IL-10 haplotype showed that the -1082G/-819C/-592C haplotype is associated with a lower risk of AD, although with a marginal statistical significance, probably due to the low number of studies included (GCC versus other genotypes: OR: 0.61, 95% CI: 0.32-1.15; I2: 85%). Current findings suggest a possible association between -1082 A > G polymorphism and the risk of developing AD; this effect is more evident in the oldest patients. The high degree of between-study heterogeneity, due to several underpowered studies and to other methodological problems of individual studies underlies the need for further methodologically adequate studies.
The cyclin kinase inhibitor p57kip2 regulates TGF-beta-induced compensatory tubular hypertrophy: effect of the immunomodulator AS101.

BACKGROUND Compensatory tubular cell hypertrophy following unilateral nephrectomy is a cell cycle-dependent process. Our previous study showed that treatment of unilaterally nephrectomized rats with the immunomodulator AS101 partially inhibits compensatory hypertrophy of the remaining kidneys through the inhibition of IL-10-induced TGF-beta secretion by mesangial cells. The present study is focused on understanding the intracellular mechanism(s) of this phenomenon. METHODS A total of 120 male Sprague-Dawley rats were unilaterally nephrectomized or sham-operated and treated with AS101 or PBS. Kidney weight and protein/DNA ratio were assessed for each experimental animal. The expression of TGF-beta, PCNA, CDK 2, pRb, ppRb, p21(Waf1), p27(kip1) and p57(kip2) proteins in renal tissues was determined by western blot analysis and immunohistochemistry, and the immunoprecipitation of cyclin E complexes was performed. RESULTS Compensatory renal growth is initiated by proliferation of resident renal cells that precedes hypertrophy. Changes in TGF-beta expression were positively correlated with the amounts of p57(kip2), but not with p21(Waf1) and p27(kip1) expression in the remaining kidneys. Moreover, there was a marked abundance of p57(kip2) but not p21(Waf1) and p27(kip1) binding to the cyclin E complex in PBS-treated unilaterally nephrectomized rats compared to sham-operated animals. Treatment of uninephrectomized rats with AS101 reduced kidney weight and protein/DNA ratio, inhibited TGF-beta and p57(kip2) expression in the remaining kidneys, and decreased the level of p57(kip2) binding to cyclin E complexes. CONCLUSION These results demonstrate that TGF-beta-induced compensatory tubular cell hypertrophy is regulated in vivo by p57(kip2) but not by the p21(Waf1) and p27(kip1) cyclin kinase inhibitor proteins.
Arginase induction promotes Trypanosoma cruzi intracellular replication in Cruzipain‐treated J774 cells through the activation of multiple signaling pathways

Given that arginase activation may effectively influence nitric oxide (NO) production in macrophages, we have investigated the intracellular signals that regulate L‐arginine metabolism and its influence on Trypanosoma cruzi growth. We demonstrate that cruzipain (Cz), a parasite antigen, induces arginase I expression in J774 cells, and the pretreatment of Cz‐treated cells with N‐omega‐hydroxy‐L‐arginine (arginase inhibitor) leads to a dramatic decrease in amastigote growth. The study of intracellular signals shows that genistein [tyrosine kinase (TK) inhibitor], KT5720 [protein kinase (PK) A inhibitor] and SB203580 [p38 mitogen‐activated protein kinase (MAPK) inhibitor] significantly decrease Cz‐induced arginase activation. However, calphostin C (PKCinhibitor) and PD98059 [p44/p42 MAPK kinase (MEK) inhibitor] did not cause a significant change. To determine if signaling pathways triggered by Cz were involved in the T. cruzi growth, westudied the effect of those inhibitors. In Cz‐treated cells – pre‐incubated with TK, PKA or p38 MAPK inhibitors – the balance of NO/urea was biased towards NO, and the amastigote growth was diminished. Besides, genistein and mainly KT5720 induced down‐regulation of arginase I expression in Cz‐treated cells. Thus, activation of TK, PKA and p38 MAPK by Cz induces an increase of arginase activity in macrophages and the subsequent T. cruzi growth.
Immunotherapy for allergic rhinitis

Allergic rhinitis (AR) affects more than 20% of the population in the United Kingdom and western Europe and represents a major cause of morbidity that includes interference with usual daily activities and impairment of sleep quality. This guidance prepared by the Standards of Care Committee (SOCC) of the British Society for Allergy and Clinical Immunology (BSACI) is for the management of AR in patients that have failed to achieve adequate relief of symptoms despite treatment with intranasal corticosteroids and/or antihistamines. The guideline is based on evidence and is for use by both adult physicians and paediatricians practising allergy. During the development of these guidelines, all BSACI members were included in the consultation process using a web‐based system. Their comments and suggestions were carefully considered by the SOCC. Where evidence was lacking, consensus was reached by the experts on the committee. Included in this guideline are indications and contraindications for immunotherapy, criteria for patient selection, the evidence for short‐ and long‐term efficacy of subcutaneous and sublingual immunotherapy, and discussion on safety and the different modes of immunotherapy including, pre‐seasonal and co‐seasonal treatments. There are sections on children, allergen standardization, vaccines used in the United Kingdom, oral allergy syndrome, cost effectiveness of immunotherapy and practical considerations of undertaking immunotherapy including recommendations on who should undertake immunotherapy and dosing schedules. Finally, there is discussion on potential biomarkers of response to immunotherapy, the use of component‐resolved diagnostics, novel approaches, alternative routes and potential areas for future research.
A mutation in the human lipoprotein lipase gene as the most common cause of familial chylomicronemia in French Canadians.

BACKGROUND Lipoprotein lipase hydrolyzes the triglyceride core of chylomicrons and very-low-density lipoproteins and has a crucial role in regulating plasma lipoprotein levels. Deficiencies of lipoprotein lipase activity lead to aberrations in lipoprotein levels. Worldwide, the frequency of lipoprotein lipase deficiency is highest among French Canadians. We sought to determine the molecular basis of the disorder in this population. METHODS The entire coding sequence of the lipoprotein lipase gene from one French Canadian patient was amplified by the polymerase chain reaction and sequenced. Exon 5 from 36 other French Canadian patients was amplified and analyzed by dot blot hybridization with allele-specific oligonucleotides. RESULTS Sequence analysis revealed a missense substitution of leucine (CTG) for proline (CCG) at residue 207 in exon 5. This mutation was found on 54 of the 74 mutant alleles (73 percent) in the patients. Studies of site-directed in vitro mutagenesis have confirmed that this mutation generates inactive lipoprotein lipase and is the cause of lipoprotein lipase deficiency. CONCLUSIONS We have identified a missense mutation at residue 207 of the lipoprotein lipase gene that is the most common cause of lipoprotein lipase deficiency in French Canadians. This mutation can be easily detected by dot blot analysis, providing opportunity for definitive DNA diagnosis of the disorder and identification of heterozygous carriers.
OPTICAL IMAGING OF RESPIRATORY NEURON ACTIVITY FROM THE DORSAL VIEW OF THE LOWER BRAINSTEM

1. We visualized respiratory‐related neuron network activity in the dorsal part of the pons and medulla of an in vitro preparation from newborn rats by optical recordings using a voltage‐sensitive dye. We measured optical signals from several seconds before to several seconds after the inspiratory phase using the inspiratory motor nerve discharge as the trigger signal and we averaged the optical signals of 20–50 respiratory cycles to obtain an optical image correlating specifically to inspiratory activity.
FGF‐2/fibroblast growth factor receptor/heparin‐like glycosaminoglycan interactions: a compensation model for FGF‐2 signaling

Heparin‐like glycosaminoglycans (HL‐GAGs) play a central role in the biological activity and signaling behavior of basic fibroblast growth factor (FGF‐2). Recent studies, however, indicate that FGF‐2 may be able to signal in the absence of HLGAG, raising the question of the nature of the role of HLGAG in FGF‐2 signaling. In this study, we present a conceptual framework for FGF‐2 signaling and derive a simple model from it that describes signaling via both HL‐GAG‐independent and HLGAG‐dependent pathways. The model is validated with F32 cell proliferation data using wild‐type FGF‐2, heparin binding mutants (K26A, K119A/R120A, K125A), and receptor binding mutants (Y103A, Y111A/W114A). In addition, this model can predict the cellular response of FGF‐2 and its mutants as a function of FGF‐2 and HLGAG concentration based on experimentally determined thermodynamic parameters. We show that FGF‐2‐mediated cellular response is a function of both FGF‐2 and HLGAG concentrations and that a reduction of one of the components can be compensated for by an increase in the other to achieve the same measure of cellular response. Analysis of the mutant FGF‐2 molecules show that reduction in heparin binding interactions and primary receptor site binding interactions can also be compensated for in the same manner. These results suggest a molecular mechanism that could be used by cells in physiological systems to modulate the FGF‐2‐mediated cellular response by controlling HLGAG expression. —Padera, R. , Venkataraman, G. , Berry, D. , Godavarti, R. , Sasisekharan, R. FGF‐2/fibroblast growth factor receptor/heparin‐like glycosaminoglycan interactions: a compensation model for FGF‐2 signaling. FASEB J. 13, 1677–1687 (1999)