How Inflammation Pathways Contribute to Cell Death in Neuro-Muscular Disorders

Neuro-muscular disorders include a variety of diseases induced by genetic mutations resulting in muscle weakness and waste, swallowing and breathing difficulties. However, muscle alterations and nerve depletions involve specific molecular and cellular mechanisms which lead to the loss of motor-nerve or skeletal-muscle function, often due to an excessive cell death. Morphological and molecular studies demonstrated that a high number of these disorders seem characterized by an upregulated apoptosis which significantly contributes to the pathology. Cell death involvement is the consequence of some cellular processes that occur during diseases, including mitochondrial dysfunction, protein aggregation, free radical generation, excitotoxicity and inflammation. The latter represents an important mediator of disease progression, which, in the central nervous system, is known as neuroinflammation, characterized by reactive microglia and astroglia, as well the infiltration of peripheral monocytes and lymphocytes. Some of the mechanisms underlying inflammation have been linked to reactive oxygen species accumulation, which trigger mitochondrial genomic and respiratory chain instability, autophagy impairment and finally neuron or muscle cell death. This review discusses the main inflammatory pathways contributing to cell death in neuro-muscular disorders by highlighting the main mechanisms, the knowledge of which appears essential in developing therapeutic strategies to prevent the consequent neuron loss and muscle wasting

Epigenetic regulation of nuclear PLCbeta1 and Cyclin D3 during Azacitidine treatment

The Myelodysplastic Syndromes (MDS) are a heterogeneous group of bone marrow disorders characterized by alterations of the hematopoietic stem cells that lead to anemia, neutropenia, bleeding problems and infections. The evidence of a clinical correlation between the presence of a monoallelic gene deletion of Phospholipase Cβ1 (PLCβ1) and the progression of MDS to Acute Myeloid Leukemia (AML) opened new perspectives of research and treatments. Patients affected by MDS with a higher risk of AML evolution have a reduction in the expression of the nuclear PLCβ1, which is also epigenetically relevant in MDS. This strengthens the importance of PLCβ1 localization. In fact, PLCβ1 is a molecular target for hypomethylating agents, such Azacitidine (AZA)(1). High-risk MDS patients that respond to the drug showed an increased expression of nuclear PLCβ1 and its downstream target Cyclin D3 (CCND3), an induction of normal myeloid differentiation, and a better prognosis. Stemming from these data, our goal was to analyze the correlation between CCND3, PLCβ1 and AZA treatment. Firstly, we treated two different cellular lines, AML HL60 and histiocytic lymphoma U937, with AZA 5μM (Ec50 for HL60 cells) for 24 hours. Then, we used Real-Time PCR and Western blot to quantify both gene and protein expression. Moreover, we showed that CCND3 promoter has one CpG island. For this reason, it is possible that AZA could directly affect both PLCβ1 and CCND3 promoters. Therefore, we studied PLCβ1 binding to CCND3 promoter by chromatin immunoprecipitation (CHIP), before and after AZA treatment. Our results evidenced that the recruitment of PLCβ1 to CCND3 promoter is specifically increased after AZA treatment, leading to suppose that PLCβ1 could have a pivotal role in MDS with either a direct or indirect effect on cell cycle, proliferation and differentiation. These complicate relations need future deepening in order to demonstrate how PLCβ1 binding actually regulates CCND3 expression and how much this expression depends on CCND3 direct promoter demethylation and PLCβ1 control

Hypo and retrotympanum: the importance of anatomical variants

The hypo- and retrotympanum host a variety of crucial anatomical structures1, characterized by high variability, which are poorly been described. The aim of our study is to describe and classify the anatomical variants of the hypo- and retrotympanum by the means of transcanal endoscopy2. We hypothesize that the retro- and hypotympanum are subject to more anatomical variability than actually thought. Moreover, the configuration as bridge variants and variably shaped sinus interconnects the different subregions. A total of 125 middle ears (83 cadaveric dissections) were explored by the means of 3mm straight and angled scopes. The variants were documented photographically and tabularized. The bony crests ponticulus, subiculum and finiculus1 were most frequently represented as ridges. The ponticulus showed the highest variability with 38% ridge, 35% bridge and 27% incomplete presentation. The subiculum was bridge - shaped only in 8% of the cases, while the finiculus in 17%. The sinus tympani had a normal shape in 66% of the cases. A subcochlear canaliculus was observed in 50%. The retro- and hypotympanum were classified respectively to the present bony crests and sinus in chambers type I to IV. In our opinion, the retro- and hypotympanum have to be considered as a tightly coherent region of the middle ear. For this purpose, we propose a straightforward classification, according to the presence of the different bony crests and sinus forming the different chambers of the retro- and hypotympanum. The introduced classification may also serve as intraoperative assessment, to be aware of the different anatomical subregions. The hidden areas of the retro- and hypotampanum are difficult to access and therefore represent a region of risk for residual cholesteatomatous disease after surgical treatment. The extension below a bridge bony crest or into a deep sinus demands thorough exploration; therefore, exact anatomical knowledge and an effective technique to visualize the whole middle ear are required