Immediate-early gene expression in human saphenous veins harvested during coronary artery bypass graft operations

Saphenous vein graft occlusion is a common late complication of coronary bypass grafting. Intimal smooth muscle cell hyperplasia is a component of this pathobiology, but the underlying molecular events are poorly understood. Immediate-early genes are activated shortly after growth stimulation and subserve cellular functions, which may contribute to intimal smooth muscle cell accumulation. In the present study, human saphenous vein grafts were harvested with minimal manipulation during coronary bypass and processed for isolation of total ribonucleic acid to examine changes in immediate-early gene expression of messenger ribonucleic acid by Northern blotting techniques. Thirty saphenous vein grafts were incubated at 4° C in Dulbecco's modified Eagle media from 30 minutes to 10 hours. The messenger ribonucleic acids for immediate-early genes c-fos and c-myc were weak or undetectable in controls but were increased (>10 times controls) within 1 hour (c-fos) and persisted for at least 6 hours (c-myc) after harvest. Our results demonstrate, for the first time in human vascular tissue, incipient immediate-early gene induction. This information may lead to molecular therapies to control saphenous vein graft disease

IgD Multiple Myeloma Paraproteinemia as a Cause of Myositis

A 48-years old man was diagnosed an IgD-k multiple myeloma (MM) at age 38 years for which he successfully underwent chemotherapy and bone marrow transplant. He then developed a graft-versus-host disease (GVHD) whose manifestations included, three years later, a polymyositis, diagnosed at muscle biopsy and successfully treated with steroids. Few months after polymyositis remission, myeloma relapsed and the patient was treated with thalidomide for six years with good remission. Soon after thalidomide suspension, MM relapsed again and the patient came to our observation for a new onset of neuromuscular symptoms. He underwent both muscle and peripheral nerve biopsy to discriminate between myositis (paraproteinemia versus GVHD), amyloidosis, and thalidomide toxicity. The first muscle biopsy showed an inflammatory pattern with necrotic fibres, macrophagical invasion (CD68 positive), rare interstitial cellular infiltrates (CD8 positive and CD4 negative), widespread anti-HLA positivity and negative antiMAC. The second muscle biopsy showed the same inflammatory pattern plus an involvement of blood vessels. Direct immunofluorescence for IgD showed diffuse positivity along the sarcolemmal in both muscle biopsies. Sural nerve biopsy demonstrated both demyelinating and axonal aspects with no inflammatory infiltrates, but positivity for HLA and MAC. Congo Red was negative in both skeletal muscle and peripheral nerve

La terapia fotodinamica: indicazioni e controindicazioni. Esperienza della Clinica Dermatologica

La terapia fotodinamica (PDT) è una forma di trattamento non chirurgico applicabile a diversi tipi di tumori non cutanei (tumori gastrointestinali, cerebrali, broncopolmonari, endometriali e vescicali) e ad alcuni tumori e lesioni pretumorali della cute. Il principio su cui si basa la PDT è quello di una reazione fotodinamica in grado di distruggere selettivamente le cellule tumorali, ovvero un processo chimico mediato dalla luce (processo foto-chimico) che prevede l’assorbimento della luce da parte di una sostanza fotosensibile e la successiva formazione di specie reattive dell’ossigeno (ROS), in grado di distruggere la cellula nella quale si sono formati. Dalla fine degli anni ’90 la PDT cutanea è divenuta una valida opzione terapeutica per il dermatologo, e dopo il 2000 è entrata a far parte delle linee guida internazionali per il trattamento dei tumori cutanei non melanocitari (NMSC) quali la cheratosi attinica (AK), il carcinoma basocellulare superficiale (BCC), e recentemente anche il morbo di Bowen (BD). La tollerabilità e i vantaggi per il paziente in confronto alle altre terapie ablative chirurgiche e non chirurgiche sono notevoli. La selettività per le cellule malate con risparmio del tessuto sano permette di avere una guarigione della lesione ulcerativa post-trattamento più rapida, di mantenere la funzione della cute, e di ottenere un risultato esteticamente molto più accettabile. Questi vantaggi sono particolarmente evidenti per le lesioni di ampie dimensioni o per lesioni multiple soprattutto al volto. Inoltre è utile per il trattamento di pazienti anziani o in scadenti condizioni generali o in terapia anticoagulante. In questo lavoro, presentiamo l’esperienza della Clinica Dermatologica della Fondazione IRCCS Policlinico San Matteo di Pavia a distanza di circa un anno dall’introduzione della PDT per la terapia dei tumori cutanei non melanocitari: sono stati trattati 29 pazienti e, tenendo conto che alcuni soggetti presentavano più di una lesione e che in uno stesso paziente potevano essere contemporaneamente presenti lesioni di natura diversa (BD, BCC e/o AK), sono stati nel complesso trattati 4 BD, 21 BCC e 52 AK. L’esperienza della Clinica Dermatologica conferma i dati riportati in letteratura sull’efficacia e i vantaggi di que-sto tipo di terapia: i risultati ottenuti sono più che soddisfacenti con una remissione clinica delle lesioni trattate maggiore del 90% del totale delle lesioni. Gli eventi collaterali sono stati scarsi e caratterizzati da dolore o bru-ciore localizzato all’area trattata e da aumento della pressione arteriosa durante la procedura. In conclusione, la PDT si è rivelata una terapia efficace, nonché una valida alternativa nel trattamento dei NMSC

Impact of rehabilitation on fatigue in post-COVID-19 patients: a systematic review and meta-analysis

The post-COVID-19 syndrome may affect patients after the COVID-19 post-acute phase. In particular, the 69% of patients reported persistent fatigue at the discharge. To date, no clear data are available regarding the most effective rehabilitative approaches for the treatment of this condition. Thus, this systematic review aimed to evaluate the rehabilitation treatment’s efficacy on fatigue in post-COVID-19 patients. We systematically searched PubMed, Scopus, and Web of Science databases to find longitudinal study designs presenting: post-COVID-19 patients as participants; a rehabilitative approach aimed to reduce post-COVID-19 syndrome as intervention; and fatigue intensity assessed through an evaluation tool that quantified the perceived exertion (i.e., fatigue severity scale, FSS; Borg Scale (BS); Borg Category Ratio 10, CR10; Checklist Individual Strength (CIS) fatigue scale; FACIT (Functional Assessment of Chronic Illness Therapy) fatigue scale). The present systematic review protocol was registered on PROSPERO (registration number CRD42021284058). Out of 704 articles, 6 studies were included. Nearly all patients showed COVID-19-related fatigue, and after the rehabilitation treatment, only 17% of subjects reported the persistency of symptoms. The overall effect size reported a −1.40 decrease in Borg Category Ratio 10 with a SE of 0.05 and a 95% CI between −1.50 and −1.30 (p < 0.001). The present systematic review and meta-analysis underlines the rehabilitation role in the fatigue reduction in patients affected by post-COVID-19 syndrome

The effects of an intronic polymorphism in TOMM40 and APOE genotypes in sporadic inclusion body myositis.

A previous study showed that, in carriers of the apolipoprotein E (APOE) genotype ε3/ε3 or ε3/ε4, the presence of a very long (VL) polyT repeat allele in "translocase of outer mitochondrial membrane 40" (TOMM40) was less frequent in patients with sporadic inclusion body myositis (sIBM) compared with controls and associated with a later age of sIBM symptom onset, suggesting a protective effect of this haplotype. To further investigate the influence of these genetic factors in sIBM, we analyzed a large sIBM cohort of 158 cases as part of an International sIBM Genetics Study. No significant association was found between APOE or TOMM40 genotypes and the risk of developing sIBM. We found that the presence of at least 1 VL polyT repeat allele in TOMM40 was significantly associated with about 4 years later onset of sIBM symptoms. The age of onset was delayed by 5 years when the patients were also carriers of the APOE genotype ε3/ε3. In addition, males were likely to have a later age of onset than females. Therefore, the TOMM40 VL polyT repeat, although not influencing disease susceptibility, has a disease-modifying effect on sIBM, which can be enhanced by the APOE genotype ε3/ε3

Effect of interchain separation on the photoinduced absorption spectra of polycarbazolyldiacetylenes

The photoinduced absorption spectra of a novel polycarbazolyldiacetylene with long aliphatic chains on the carbazolyl side groups are measured and compared with those of the unsubstituted polyDCHD. The two polymers in the blue form exhibit very similar electronic absorption spectra and Raman frequencies. This fact indicates that the conjugation length of the polydiacetylene backbone is not too affected by the long substituents. In contrast, the near steady-state photoinduced absorption spectra show that different photogeneration mechanisms are involved in the two polymers. This result can be ascribed to the role played by the interchain distance in the dynamics of the relaxation processes in polydiacetylenes

In vitro and in vivo tetracycline-controlled myogenic conversion of NIH-3T3 cells: evidence of programmed cell death after muscle cell transplantation.

Ex vivo gene therapy of Duchenne muscular dystrophy based on autologous transplantation of genetically modified myoblasts is limited by their premature senescence. MyoD-converted fibroblasts represent an alternative source of myogenic cells. In this study the forced MyoD-dependent conversion of murine NIH-3T3 fibroblasts into myoblasts under the control of an inducible promoter silent in the presence of tetracycline was evaluated. After tetracycline withdrawal this promoter drives the transcription of MyoD in the engineered fibroblasts, inducing their myogenesis and giving rise to β-galactosidase-positive cells. MyoD-expressing fibroblasts withdrew from the cell cycle, but were unable to fuse in vitro into multinucleated myotubes. Five days following implantation of engineered fibroblasts in muscles of C57BL/10J mice we observed a sevenfold increase of β-galactosidase-positive regenerating myofibers in animals not treated with antibiotic compared with treated animals. After 1 week the number of positive fibers decreased and several apoptotic myonuclei were detected. Three weeks following implantation of MyoD-converted fibroblasts in recipient mice, no positive "blue" fiber was observed. Our results suggest that transactivation by tetracycline of MyoD may drive an in vivo myogenic conversion of NIH-3T3 fibroblasts and that, in this experimental setting, apoptosis plays a relevant role in limiting the efficacy of engineered fibroblast transplantation. This work opens the question whether apoptotic phenomena also play a general role as limiting factors of cellmediated gene therapy of inherited muscle disorders

KISS1R and ANKRD31 Cooperate to Enhance Leydig Cell Gene Expression via the Cytoskeletal-Nucleoskeletal Pathway

Kisspeptins are involved in the regulation of hypothalamic-pituitary-gonadal axis, Leydig cell functions, and testosterone secretion, acting as endogenous ligands of the KISS1 receptor. ANKRD31 protein participates in male fertility, regulating meiotic progression, and epididymal sperm maturation. Here, we show that in Leydig cells, KISS1 receptor and ANKRD31 proteins physically interact; the formation of this protein complex is enhanced by Kisspeptin-10 that also modulates F-actin synthesis, favoring histone acetylation in chromatin and gene expression via the cytoskeletal–nucleoskeletal pathway. Kp/KISS1R system deregulation, expression impairment of cytoskeletal–nucleoskeletal mediators, Leydig gene targets, and the decreased testosterone secretion in Ankrd31−/− testis strongly supported our hypothesis. Furthermore, cytochalasin D treatment subverted the gene expression induction dependent on Kisspeptin-10 action. In conclusion, the current work highlights a novel role for the Kisspeptin-10 in the induction of the cytoskeletal–nucleoskeletal route, downstream a physical interaction between KISS1 receptor and ANKRD31, with gene expression activation as final effect, in Leydig cells

Long-lived photoexcited states in polydiacetylenes with different molecular and supramolecular organization

With the aim of determining the importance of the molecular and supramolecular organization on the excited states of polydiacetylenes, we have studied the photoinduced absorption spectra of the red form of poly[1,6-bis(3,6-didodecyl-N-carbazolyl)-2,4-hexadiyne] (polyDCHD-S) and the results compared with those of the blue form of the same polymer. An interpretation of the data is given in terms of both the conjugation length and the interbackbone separation also in relation to the photoinduced absorption spectra of both blue and red forms of poly[1,6-bis(N-carbazolyl)-2,4-hexadiyne] (polyDCHD), which does not carry the alkyl substituents on the carbazolyl side groups. Information on the above properties is derived from the analysis of the absorption and Raman spectra of this class of polydiacetylenes