Optimization of organotypic cultures of mouse spleen for staining and functional assays

By preserving cell viability and three-dimensional localization, organotypic culture stands out among the newest frontiers of cell culture. It has been successfully employed for the study of diseases among which neoplasias, where tumoral cells take advantage of the surrounding stroma to promote their own proliferation and survival. Organotypic culture acquires major importance in the context of the immune system, whose cells cross-talk in a complex and dynamic fashion to elicit productive responses. However, organotypic culture has been as yet poorly developed for and applied to primary and secondary lymphoid organs. Here we describe in detail the development of a protocol suitable for the efficient cutting of mouse spleen, which overcomes technical difficulties related to the peculiar organ texture, and for optimized organotypic culture of spleen slices. Moreover, we used microscopy, immunofluorescence, flow cytometry and qRT-PCR to demonstrate that the majority of cells residing in spleen slices remain alive and maintain their original location in the organ architecture for several days after cutting. The development of this protocol represents a significant technical improvement in the study of the lymphoid microenvironment in both physiological and pathological conditions involving the immune system

Improving sleep quality in cancer patients. a literature review on non-pharmacologic interventions

Objective: Disrupted sleep-in cancer patients is due to several factors. Pre-existing sleep disorders, mental status in cancer, or side effects of cancer treatment are all potential predictors of sleep disturbances in this vulnerable population. This review aims to evaluate and synthesise non-pharmaco-logic interventions for improving sleep quality and the associated benefits in cancer patients. Materials and Methods: A literature review was performed according to PRISMA guidelines. PubMed, Google Scholar, MEDLINE, CINAHL, and Embase databases were interrogated, selected the article with cancer, neoplasm, oncology, sleep-wake disorders, sleep disturbance, sleep problem, strategies, treat-ment, and intervention such as keywords. No time and geographic restrictions but paediatrics, children, and interventions unrelated to sleep improvement in cancer were excluded. Results: Overall, twenty-nine articles were included in the review after selection. The included studies analysed different types of cancer, like breast, lung, and prostate cancer and acute leukaemia. Most of the data gathered from the relevant research suggest that nonpharmacologic interventions significantly improved cancer patients’ sleep quality. Conclusions: Cancer survival rates are increasing; researchers and healthcare professionals should aim not just at survival but also to allow cancer patients just comfortably to live, considering the best quality of life possible. Nurses have a crucial role in the care of this patient population. Nursing implies an intimate relationship with patients and educating the patient to improve their mental and physical condition through non-pharmacological approaches, which should be considered a specific competence

Elderly patients with multimorbidity in the home setting: umbrella review on therapeutic non-adherence causes

Objective: The elderly population is the most at risk regarding adherence, especially in the coexistence of multiple diseases. This study aims to detect factors contributing to therapeutic non-adherence in elderly patients in home settings. Materials and methods: A review protocol was developed to conduct the umbrella review using the methodological framework of the Richardson et al study. The search strategy was developed in December 2022 to conduct a systematic search and to perform an Umbrella Review of systematic reviews, meta-analyses and integrative reviews published from 2012 to 2022 in English. Results: A total of 26,038 articles were identified and screened. 18 relevant articles were included in the study. Conclusions: Therapeutic adherence in elderly patients with comorbidities in polypharmacotherapy at home is a significant problem in public health and health care. Several factors of non-adherence have been identified in the studies reviewed, confirming that the problem is multifactorial. Reducing the number of medications prescribed would appear optimal, although often not possible, as this has been seen to have an immediate positive impact. A multidisciplinary approach makes it possible not to fragment care, ensuring positive feedback on therapeutic adherence

Improving sleep quality in cancer patients: a literature review on non-pharmacologic interventions

Objective: Disrupted sleep-in cancer patients is due to several factors. Pre-existing sleep disorders, mental status in cancer, or side effects of cancer treatment are all potential predictors of sleep disturbances in this vulnerable population. This review aims to evaluate and synthesise non-pharmacologic interventions for improving sleep quality and the associated benefits in cancer patients. Materials and Methods: A literature review was performed according to PRISMA guidelines. PubMed, Google Scholar, MEDLINE, CINAHL, and Embase databases were interrogated, selected the article with cancer, neoplasm, oncology, sleep-wake disorders, sleep disturbance, sleep problem, strategies, treatment, and intervention such as keywords. No time and geographic restrictions but paediatrics, children, and interventions unrelated to sleep improvement in cancer were excluded. Results: Overall, twenty-nine articles were included in the review after selection. The included studies analysed different types of cancer, like breast, lung, and prostate cancer and acute leukaemia. Most of the data gathered from the relevant research suggest that nonpharmacologic interventions significantly improved cancer patients' sleep quality. Conclusions: Cancer survival rates are increasing; researchers and healthcare professionals should aim not just at survival but also to allow cancer patients just comfortably to live, considering the best quality of life possible. Nurses have a crucial role in the care of this patient population. Nursing implies an intimate relationship with patients and educating the patient to improve their mental and physical condition through non-pharmacological approaches, which should be considered a specific competence

Multilocus genotyping reveals new molecular markers for differentiating distinct genetic lineages among “candidatus phytoplasma solani” strains associated with grapevine bois noir

Grapevine Bois noir (BN) is associated with infection by “Candidatus Phytoplasma solani” (CaPsol). In this study, an array of CaPsol strains was identified from 142 symptomatic grapevines in vineyards of northern, central, and southern Italy and North Macedonia. Molecular typing of the CaPsol strains was carried out by analysis of genes encoding 16S rRNA and translation elongation factor EF-Tu, as well as eight other previously uncharacterized genomic fragments. Strains of tuf-type a and b were found to be differentially distributed in the examined geographic regions in correlation with the prevalence of nettle and bindweed. Two sequence variants were identified in each of the four genomic segments harboring hlyC, cbiQ-glyA, trxA-truB-rsuA, and rplS-tyrS-csdB, respectively. Fifteen CaPsol lineages were identified based on distinct combinations of sequence variations within these genetic loci. Each CaPsol lineage exhibited a unique collective restriction fragment length polymorphism (RFLP) pattern and differed from each other in geographic distribution, probably in relation to the diverse ecological complexity of vineyards and their surroundings. This RFLP-based typing method could be a useful tool for investigating the ecology of CaPsol and the epidemiology of its associated diseases. Phylogenetic analyses highlighted that the sequence variants of the gene hlyC, which encodes a hemolysin III-like protein, separated into two clusters consistent with the separation of two distinct lineages on the basis of tufB gene sequences. Alignments of deduced full protein sequences of elongation factor-Tu (tufB gene) and hemolysin III-like protein (hlyC gene) revealed the presence of critical amino acid substitutions distinguishing CaPsol strains of tuf-type a and b. Findings from the present study provide new insights into the genetic diversity and ecology of CaPsol populations in vineyards

CSN-mediated deneddylation differentially modulates Ci155 proteolysis to promote Hedgehog signalling responses

The Hedgehog (Hh) morphogen directs distinct cell responses according to its distinct signalling levels. Hh signalling stabilizes transcription factor cubitus interruptus (Ci) by prohibiting SCFSlimb-dependent ubiquitylation and proteolysis of Ci. How graded Hh signalling confers differential SCFSlimb-mediated Ci proteolysis in responding cells remains unclear. Here, we show that in COP9 signalosome (CSN) mutants, in which deneddylation of SCFSlimb is inactivated, Ci is destabilized in low-to-intermediate Hh signalling cells. As a consequence, expression of the low-threshold Hh target gene dpp is disrupted, highlighting the critical role of CSN deneddylation on low-to-intermediate Hh signalling response. The status of Ci phosphorylation and the level of E1 ubiquitin-activating enzyme are tightly coupled to this CSN regulation. We propose that the affinity of substrate–E3 interaction, ligase activity and E1 activity are three major determinants for substrate ubiquitylation and thereby substrate degradation in vivo

Efficacia clinica della roxitromicina in pazienti affetti da infezioni odontogene acute

The clinical efficacy of a macrolide antibiotic, roxithromycin, was evaluated in 24 patients affected by acute odontogenic infections. Patients were treated with a first dose of 300 mg p.o. which was followed by 150 mg p.o. 12-hourly for the following two days. Total responses were evaluated by an arbitrary scale. The results showed that an excellent and good response was obtained in 99.96% of treated patients. On the basis of clinically compared data and the drug safety it may be concluded that roxithromycin can be successfully used in the treatment of odontogenic infections