p73 Is Regulated by Phosphorylation at the G2/M Transition *

p73 is a p53 paralog that encodes proapoptotic (transactivation-competent (TA)) and antiapoptotic (dominant negative) isoforms. TAp73 transcription factors mediate cell cycle arrest and/or apoptosis in response to DNA damage and are involved in developmental processes in the central nervous system and the immune system. p73 proteins may also play a role in the regulation of cell growth. Indeed, p73 expression is itself modulated during the cell cycle and TAp73 proteins accumulate in S phase cells. In addition, the function of p73 proteins is also regulated by post-translational modifications and protein-protein interactions in different cellular and pathophysiological contexts. Here we show that p73 is a physiological target of the p34cdc2-cyclin B mitotic kinase complex in vivo. Both p73beta and p73alpha isoforms are hyperphosphorylated in normal mitotic cells and during mitotic arrest induced by microtubule-targeting drugs. p34cdc2-cyclin B phosphorylates and associates with p73 in vivo, which results in a decreased ability of p73 to both bind DNA and activate transcription in mitotic cells. Indeed, p73 is excluded from condensed chromosomes in meta- and anaphase, redistributes throughout the mitotic cytoplasm, and unlike p53, shows no association with centrosomes. Together these results indicate that M phase-specific phosphorylation of p73 by p34cdc2-cyclin B is associated with negative regulation of its transcriptional activating function

Reactive oxygen intermediates mediate angiotensin II-induced c-Jun.c-Fos heterodimer DNA binding activity and proliferative hypertrophic responses in myogenic cells

Angiotensin II (Ang-II) receptor engagement activates many immediate early response genes in both vascular smooth muscle cells and cardiomyocytes whether a hyperplastic or hypertrophic response is taking place. Although the signaling pathways stimulated by Ang-II in different cell lines have been widely characterized, the correlation between the generation of different second messengers and specific physiological responses remains relatively unexplored. In this study, we report how in both C2C12 quiescent myoblasts and differentiated myotubes Ang-II significantly stimulates AP1-driven transcription and c-Jun.c-Fos heterodimer DNA binding activity. Using a set of different protein kinase inhibitors, we could demonstrate that Ang-II-induced increase in AP1 binding is not mediated by the cAMP-dependent pathway and that both protein kinase C and tyrosine kinases are involved. The observation that in quiescent myoblasts Ang-II increase of AP1 binding and induction of DNA synthesis and, in differentiated myotubes, Ang-II stimulation of protein synthesis are abolished by the cysteine-derivative and glutathione precursor N-acetyl-L-cysteine strongly suggests a role for reactive oxygen intermediates in the intracellular transduction of Ang-II signals for immediate early gene induction, cell proliferation, and hypertrophic responses

The efficacy of blended learning in a Pediatric Spine Deformity management program in Sub-Saharan Africa

Introduction: Our study assessed the efficacy of blended learning, which combines in-person learning and e-learning, in a pediatric scoliosis training program through an international collaborative effort. Methods: The course comprised two parts: the online portion, where participants reviewed educational materials for 3 weeks and met with faculty once/week for discussion, and the in-person session, where participants reviewed cases in a team-based approach and came to a consensus on treatment strategy, followed by discussion with an international expert. All participants completed a needs assessment (NA) and clinical quiz at three points: before the course, after the online session, and after the in-person session, which covered various topics in pediatric spine deformity. Results: Thirty-six surgeons enrolled in the course from 13 College of Surgeons of East, Central and Southern Africa countries. The NA assessment scores improved significantly over the course of the surveys from 67.3, to 90.9, to 94.0 (P = 0.02). The clinical quiz scores also improved from 9.91, to 11.9, to 12.3 (P = 0.002). Conclusion: The blended learning approach in a pediatric spine deformity program is effective and feasible and shows a statistically significant change in participants\u27 confidence and knowledge base in these complex pathologies. This approach should be explored further with larger numbers and/or other spinal pathologies

Myosteatosis is closely associated with sarcopenia and significantly worse outcomes in patients with cirrhosis

Background & aims: Sarcopenia and myosteatosis are common in patients with cirrhosis. This study aimed to determine the prevalence of these muscle changes, their interrelations and their prognostic impact over a 12-month period. Methods: We conducted a prospective multicentre study involving 433 patients. Sarcopenia and myosteatosis were evaluated using computed tomography scans. The 1-year cumulative incidence of relevant events was assessed by competing risk analysis. We used a Fine-Gray model adjusted for known prognostic factors to evaluate the impact of sarcopenia and myosteatosis on mortality, hospitalization, and liver decompensation. Results: At enrolment, 166 patients presented with isolated myosteatosis, 36 with isolated sarcopenia, 135 with combined sarcopenia and myosteatosis and 96 patients showed no muscle changes. The 1-year cumulative incidence of death in patients with either sarcopenia and myosteatosis (13.8%) or isolated myosteatosis (13.4%) was over twice that of patients without muscle changes (5.2%) or with isolated sarcopenia (5.6%). The adjusted sub-hazard ratio for death in patients with muscle changes was 1.36 (95% CI 0.99-1.86, p = 0.058). The cumulative incidence of hospitalization was significantly higher in patients with combined sarcopenia and myosteatosis than in patients without muscle changes (adjusted sub-hazard ratio 1.18, 95% CI 1.04-1.35). The cumulative incidence of liver decompensation was greater in patients with combined sarcopenia and myosteatosis (p = 0.018) and those with isolated sarcopenia (p = 0.046) than in patients without muscle changes. Lastly, we found a strong correlation of function tests and frailty scores with the presence of muscle changes. Conclusions: Myosteatosis, whether alone or combined with sarcopenia, is highly prevalent in patients with cirrhosis and is associated with significantly worse outcomes. The prognostic role of sarcopenia should always be evaluated in relation to the presence of myosteatosis. Impact and implications: This study investigates the prognostic role of muscle changes in patients with cirrhosis. The novelty of this study is its multicentre, prospective nature and the fact that it distinguishes between the impact of individual muscle changes and their combination on prognosis in cirrhosis. This study highlights the prognostic role of myosteatosis, especially when combined with sarcopenia. On the other hand, the relevance of sarcopenia could be mitigated when considered together with myosteatosis. The implication from these findings is that sarcopenia should never be evaluated individually and that myosteatosis may play a dominant role in the prognosis of patients with cirrhosis

SPINE20 recommendations 2021: spine care for people’s health and prosperity

PurposeThe focus of SPINE20 is to develop evidence-based policy recommendations for the G20 countries to work with governments to reduce the burden of spine disease, and disability.MethodsOn September 17-18, 2021, SPINE20 held its annual meeting in Rome, Italy. Prior to the meeting, the SPINE20 created six proposed recommendations. These recommendations were uploaded to the SPINE20 website 10 days before the meeting and opened to the public for comments. The recommendations were discussed at the meeting allowing the participants to object and provide comments.ResultsIn total, 27 societies endorsed the following recommendations. SPINE20 calls upon the G20 countries: (1) to expand telehealth for the access to spine care, especially in light of the current situation with COVID-19. (2) To adopt value-based interprofessional spine care as an approach to improve patient outcomes and reduce disability. (3) To facilitate access and invest in the development of a competent rehabilitation workforce to reduce the burden of disability related to spine disorders. (4) To adopt a strategy to promote daily physical activity and exercises among the elderly population to maintain an active and independent life with a healthy spine, particularly after COVID-19 pandemic. (5) To engage in capacity building with emerging countries and underserved communities for the benefit of spine patients. (6) To promote strategies to transfer evidence-based advances into patient benefit through effective implementation processes.ConclusionsSPINE20's initiatives will make governments and decision makers aware of efforts to reduce needless suffering from disabling spine pain through education that can be instituted across the globe

Sustained activation of the Raf/MEK/Erk pathway in response to EGF in stable cell lines expressing the hepatitis C virus (HCV) core protein

Chronic hepatitis C virus (HCV) infection is a leading cause of liver cirrhosis and hepatocellular carcinoma (HCC) worldwide. The HCV capside core is a multifunctional protein with regulatory functions that affects transcription and cell growth in vitro and in vivo. Here, we show that both HCV genotype 1a and 3 core proteins activate MEK1 and Erk1/2 MAP kinases and that the costitutive expression of the HCV core results in a high basal activity of Raf1 and MAP/kinase/kinase, as determined by endogenous Raf1 in vitro kinase assay and immunodetection of hyperphosphorylated Erk1 and Erk2 even after a serum starvation. Moreover, the activation of both Erk1/2 and the downstream transcription factor Elk-1 in response to the mitogenic stimulus EGF is significantly prolonged. The sustained response to EGF in cells expressing the HCV core occurs despite a normal induction of the MAP phosphatases MKP regulatory feedback and is likely due to the costitutive activation of Raf-1 activity. The ability of HCV core proteins to directly activate the MAP kinase cascade and to prolong its activity in response to mitogenic stimuli may contribute to the neoplastic transformation of HCV infected liver cells

Vertebral body tethering: An alternative to posterior spinal fusion in idiopathic scoliosis?

IntroductionSkeletally immature patient with adolescent idiopathic scoliosis (AIS) whose curves continue to progress despite bracing should be treated surgically. Vertebral body tethering (VBT) is a non-fusion, compression-based, growth preserving alternative to posterior spinal fusion (PSF) based on the concept of ‘growth modulation’ to prevent possible functional complications secondary to fusion while correcting scoliotic deformity. This review aims to shed light on the indications of VBT, short- and medium-term outcomes, describe the surgical technique and associated complications, and to compare its efficacy to that of PSF.MethodsA review of peer-reviewed literature on VBT as a surgical technique, its indications, outcomes, complications, and comparison with other surgical interventions to correct AIS was conducted in December 2022.ResultsIndications remain controversial and mainly include stage of skeletal maturity based on radiographic markers, curve location, magnitude and flexibility, and presence of secondary curve. Assessment of VBT clinical success should not be restricted to improvement in radiographic parameters but should include functional results and patient-centered outcomes, improved body image and pain, and durability of outcomes. In contrast to fusion, VBT seems to be associated with preserved spinal growth, shorter recovery, potentially better functional outcomes, less motion loss but possibly less curve correction.DiscussionYet still, with VBT there exists a risk of overcorrection, construct breakage or failure of procedure which require revision and at times conversion to PSF. Patient and family preferences must be accounted for acknowledging gaps in knowledge, attributes and drawbacks of each intervention

INVITRO INDUCTION OF HBSAG-SPECIFIC CD8 CD11 HUMAN SUPPRESSOR T-CELLS

Anti-hepatitis B surface (HBs) antibodies have been induced in vitro by human peripheral blood mononuclear cells (PBMC) from individuals immunized with hepatitis B surface antigen (HBsAg). Anti-HBs antibody production is antigen-specific, T-cell dependent, class II MHC-restricted and requires de novo synthesis. Furthermore, HBsAg-specific CD8 suppressor cells can also be induced in vitro after challenge with high antigen doses. Antigen presenting cells (APC) are required for the induction of suppression. These suppressor cells are antigen-specific since they do not suppress the antibody response to tetanus toxoid. Antigen-specific suppression is inhibited by cytotoxic treatment of CD8 CD11 cells with OKM1 monoclonal antibody (MoAb) and complement, suggesting that these suppressor CD8 cells may represent granular lymphocytes. Addition to cultures of high concentrations of a recombinant human IL-2 does not affect suppression, ruling out the adsorption of IL-2 by suppressor cells as a possible mechanism for suppression

Immunoregulatory T cells in HBV-induced chronic liver disease as defined by monoclonal antibodies

Peripheral blood T lymphocytes and T-cell subsets were analyzed in 19 patients with HBV-associated chronic liver disease, in 9 "autoimmune" chronic active hepatitis patients, and in three patients with HBV acute hepatitis. The percentages of the different T-cell subpopulations were defined by indirect immunofluorescence using monoclonal antibodies against all peripheral blood T cells (OKT3), T helper cells (OKT4), T suppressor cytotoxic cells (OKT8), and Ia antigens (OKIa1). The OKT4/OKT8 ratio was significantly lower in HBsAg+ chronic liver disease patients as compared with controls (P < 0.001), with both decreases of T helper cells and increases of T suppressor cells while "autoimmune" chronic active hepatitis patients showed a significantly increased ratio (P< 0.001). Acute HBV infection was associated with a reversal in the normal ratio of helper to suppressor T lymphocytes. During convalescence helper T lymphocytes increased and suppressor T lymphocytes decreased. © 1983

Hepatitis B virus X protein transactivates the long terminal repeats of human immunodeficiency virus types 1 and 2

The X gene product of the hepatitis B virus (HBV) has been expressed transiently in HepG2 cells, and the 17-kilodalton protein has been detected by Western (immuno-) blot analysis. Cotransfection of the X gene with the long terminal repeat of human immunodeficiency virus type 1 or 2 results in a stimulation of long terminal repeat-directed expression that is higher than the X-induced stimulation of the HBV enhancer linked to either autologous promoter or to the heterologous simian virus 40 promoter. A frameshift mutation abolished this transactivation. In vitro nuclear transcription assays revealed that HBV X acts at the transcriptional level. The carboxy terminus of the HBV X protein does not seem to be necessary for its transactivating activity, as demonstrated by using HBV X protein deletion mutants