Post-acute cardiac complications following SARS-CoV-2 infection in partial lipodystrophy due to LMNA gene p.R349W mutation.

PURPOSE: SARS-CoV-2 infection may cause varying degrees of cardiac injury and the presence of underlying cardiovascular morbidities contributes to the frequency and severity of occurrence of this complication. Lipodystrophy syndromes are frequently characterized by severe metabolic derangements that represent relevant cardiovascular risk factors. Besides causing lipodystrophy, mutations in the lamin A/C (LMNA) gene can lead to a wide spectrum of tissue-specific disorders including cardiac involvement. METHODS AND RESULTS: We herein examine the case of two patients affected by atypical progeroid syndrome and partial lipodystrophy due to a heterozygous missense LMNA mutation c.1045 C > T (p.R349W) who presented initially with mild COVID-19 and developed severe cardiovascular complications within few weeks of SARS-CoV-2 infection. Before being infected with SARS-CoV-2, our patients had cardiovascular morbidities (mild mitral regurgitation in one patient, ischemic heart disease with bifascicular block in the other patient) in adjunct to cardiovascular risk factors, but the SARS-CoV-2 infection contributed to quickly and significantly decompensate their balance. CONCLUSION: These findings warn that patients affected by LMNA p.R349W mutation and likely other LMNA mutations associated with cardiovascular morbidity should be considered at extremely elevated risk of post-acute cardiological manifestations and should therefore undergo a vigilant follow-up after SARS-CoV-2 infection. Both patients developed COVID-19 before the specific vaccination was available to them and this unfortunate situation should remark the importance of vaccination coverage against SARS-CoV-2 infection for all patients affected by lipodystrophy, especially those with underlying comorbidities.

Large-scale phylogenomic analysis reveals the phylogenetic position of the problematic taxon Protocruzia and unravels the deep phylogenetic affinities of the ciliate lineages.

The Ciliophora is one of the most studied protist lineages because of its important ecological role in the microbial loop. While there is an abundance of molecular data for many ciliate groups, it is commonly limited to the 18S ribosomal RNA locus. There is a paucity of data when it comes to availability of protein-coding genes especially for taxa that do not belong to the class Oligohymenophorea. To address this gap, we have sequenced EST libraries for 11 ciliate species. A supermatrix was constructed for phylogenomic analysis based on 158 genes and 42,158 characters and included 16 ciliates, four dinoflagellates and nine apicomplexans. This is the first multigene-based analysis focusing on the phylum Ciliophora. Our analyses reveal two robust superclades within the Intramacronucleata; one composed of the classes Spirotrichea, Armophorea and Litostomatea (SAL) and another with Colpodea and Oligohymenophorea. Furthermore, we provide corroborative evidence for removing the ambiguous taxon Protocruzia from the class Spirotrichea and placing it as incertae sedis in the phylum Ciliophora.

Identification and initial characterization of an autocrine pheromone receptor in the protozoan ciliate Euplotes raikovi.

The polypeptide pheromone Er-1, purified from the ciliate Euplotes raikovi of mating type I and genotype mat-1/mat-1, was iodinated with 125I-Bolton-Hunter reagent to a sp act of 0.45-0.73 mu Ci/microgram of protein. This preparation of 125I-Er-1 bound specifically to high affinity binding sites on the same cells of mating type I. Binding of 125I-Er-1 occurred with an apparent Kd of 4.63 +/- 0.12 X 10(-9) M in cells in early stationary phase. It was estimated that these cells carry a total number of approximately 5 X 10(7) sites/cell, with a site density that falls in the range of 1,600-1,700/microns 2 of cell surface. Unlabeled Er-1, other homologous pheromones such as Er-2 and Er-10, antibodies specific for Er-1, and human IL-2 were shown to act as effective inhibitors of specific binding of 125I-Er-1 to mating type I cells. The "autocrine" nature of the identified specific high affinity binding sites for Er-1 was further substantiated by cross-linking experiments. These experiments revealed that mating type-I cell membranes contain one protein entity of Mr = 28,000 that is capable of reacting specifically with the homodimeric native form of Er-1.

Epigenetic Changes in Chronic Inflammatory Diseases.

The number of people diagnosed with chronic inflammatory diseases has increased noteworthy in the last 40 years. Spondyloarthritis (SpA), inflammatory bowel diseases (IBD), and psoriasis are the most frequent chronic inflammatory diseases, resulting from a combination of genetic predisposition and environmental factors. Epigenetic modifications include DNA methylation, histone modifications, and small and long noncoding RNAs. They are influenced by environmental exposure, life-style, and aging and have recently been shown to be altered in many complex diseases including inflammatory diseases. While epigenetic modifications have been well characterized in other diseases such as cancer and autoimmune diseases, knowledge on changes in inflammatory diseases is lagging behind with some disease-specific differences. While the DNA methylation profile of different cell types in patients with IBD has been relatively well described, less is known on changes implicated in psoriasis, and no systematic genome-wide studies have so far been performed in SpA. In this chapter, we review in detail the reported changes in patterns of DNA methylation and posttranslational histone modifications in chronic inflammatory diseases highlighting potential connections between disease-associated pathophysiological changes such as the dysbiosis of the microbiome or genetic variations associated with disease susceptibility and the epigenome. We also discuss important parameters of meaningful epigenetic studies such as the use of well defined, disease-relevant cell populations, and elude on the potential future of engineering of the epigenome in inflammatory diseases.

Cadmium metallothionein gene of Tetrahymena pyriformis.

A genomic sequence from Tetrahymena pyriformis, encoding a cadmium-induced metallothionein has been cloned. The gene encodes a transcript of 487 bases, with an intronless coding region of 324 nt, using TGA as the stop codon, TAA coding for glutamine, and the translational initiation sequence AAAATGG. Two regions of internal similarity in the coding sequence support the hypothesis that the Tetrahymena protein arose by gene duplication. The sequences of untranslated regions show some similarities with nematode MT-1 and MT-2 transcripts. Sequence of 525 bases upstream of the transcription start contains a TATA box, a CAAT box reverse complement, and many short stretches partially matching the AP-1 and ACE-1 binding sites, but no characteristic sequences found in other metallothionein promoters.

Long-term culture and characterization of alloreactive T-cell infiltrates from renal needle biopsies.

Long-term T-cell lines have been established by culturing renal needle biopsies from kidney recipients undergoing graft rejection. These cultures are maintained in the presence of Interleukin 2, with weekly addition of irradiated donor lymphocytes as a source of antigen. The cells express T-cell markers, T3, T4, and T8, T-cell growth factor (IL2) receptor, and HLA-DR antigens. In one cell line, J2, the T4 and T8 antigens are found on two separate cell populations. This line is also shown to specifically kill Epstein Barr Virus transformed donor cells. This cytotoxicity is directed against HLA-B35 determinant and has been maintained in vitro for over 6 months.

Tumour necrosis factor-alpha blockers in rheumatoid arthritis: review of the clinical experience.

Tumour necrosis factor (TNF)-alpha has been found to play a central role in the pathogenesis of rheumatoid arthritis, leading to development of novel drug therapies that neutralise the deleterious effects of this cytokine. This new concept of immunobiological treatment of rheumatoid arthritis has yielded successful results. Although the 2 currently available TNFalpha blockers, infliximab and etanercept, differ in structure, mechanism of action and pharmacokinetics, they have provided similar benefits both in clinical improvement and in slowing and even arresting the progression of radiographic damage. This therapeutic response seems to be unequalled by "conventional" treatments in rheumatoid arthritis, and is incontestably a turning point in the therapeutic management of this disease.

Development of the Abbott AxSYM Free PSA assay: performance characteristics and preliminary clinical evaluation.

The AxSYM Free PSA assay was demonstrated to have good analytical sensitivity and reproducibility. The F/T ratio determinations for 385 men tested during the Prostate Awareness Week who had biopsies due to an elevated total PSA value and/or a suspicious DRE demonstrated that the percentage of free PSA was lower in patients found to have prostate cancer than those that were biopsy negative for the overall group and for all patient categories examined. The optimal strategy for combining PSA values, F/T ratios, DRE and other clinical and diagnostic parameters to improve the early detection of prostate cancer requires additional clinical studies.

The ethogram of Euplotes crassus (ciliata, hypotrichida): I. The wild type.

The behavior of Euplotes crassus has been studied qualitatively and quantitatively in an attempt of throwing new light onto the problem of the relationships occurring among motor behavior, systematic position and evolutionary strategies of the ciliated protozoa. It was found that both single and paired Euplotes creep along piecewise pathways whose geometric elements (arcs, segments, angles) have been measured. E. crassus performs different delicate maneuvers to shift from one element of its trajectory to the next: beyond the Continuous Trajectory Change, the Smooth Trajectory Change and the Rough Trajectory Change, described elsewhere for other ciliates, three different kinds of Side-Stepping Reaction were recognized and carefully described. The average velocity is 850 µm/sec during creeping and 800 µm/sec during swimming. Finally, also the mating behavior has been analyzed and the successive steps leading to pair formation thoroughly described. The data are compared with those already available for different species.

Self from nonself discrimination in single-celled eukaryotes.

A mechanism for self recognition has been proposed to control the mating type interactions in the marine ciliate Euplotes raikovi. The relevant molecules involved in this mechanism are the mating pheromones inherited via high-multiple alleles codominant at the Mendelian locus mat. Four of these mating pheromones have been isolated, purified, and characterized. They are relatively small and acidic proteins (Mr, 11,000-12,000; pI, 3.7-4.0), which are still active at the concentration of approximately 10(-12) molar. Data, yet preliminary, of amino acid sequencing of the purified mating pheromones showed extensive structural homologies and the common presence of the aspartic acid at the amino terminal.

Cell cycle inhibition therapy that targets stathmin in in vitro and in vivo models of breast cancer.

Stathmin is the founding member of a family of microtubule-destabilizing proteins that have a critical role in the regulation of mitosis. Stathmin is expressed at high levels in breast cancer and its overexpression is linked to disease progression. Although there is a large body of evidence to support a role for stathmin in breast cancer progression, the validity of stathmin as a viable therapeutic target for breast cancer has not been investigated. Here, we used a bicistronic adenoviral vector that co-expresses green fluorescent protein and a ribozyme that targets stathmin messenger RNA in preclinical breast cancer models with different estrogen receptor (ER) status. We examined the effects of anti-stathmin ribozyme on the malignant phenotype of breast cancer cells in vitro and in xenograft models in vivo both as a single agent and in combination with chemotherapeutic agents. Adenovirus-mediated gene transfer of anti-stathmin ribozyme resulted in a dose-dependent inhibition of proliferation and clonogenicity associated with a G2/M arrest and increase in apoptosis in both ER-positive and ER-negative breast cancer cell lines. This inhibition was markedly enhanced when stathmin-inhibited breast cancer cells were exposed to low concentrations of taxol, which resulted in virtually complete loss of the malignant phenotype. Interestingly, breast cancer xenografts treated with low doses of anti-stathmin therapy and taxol showed regression in a majority of tumors, while some tumors stopped growing completely. In contrast, combination of anti-stathmin ribozyme and adriamycin resulted in only a modest inhibition of growth in vitro and in breast cancer xenografts in vivo. Although inhibition of tumor growth was observed in both the combination treatment groups compared with groups treated with single agent alone, combination of anti-stathmin therapy and taxol had a more profound inhibition of tumorigenicity, as both agents target the microtubule pathway. Clinically, these findings are highly relevant because taxol is one of the most active chemotherapeutic agents in breast cancer. These studies provide the proof-of-principle that stathmin provides an attractive molecular target, which could serve as a primary focus of novel approaches to breast cancer.

Isolation of the mating-inducing factor of the ciliate Euplotes.

Numerous strains of different mating types of the marine ciliate Euplotes raikovi have been found to be autonomous excreters into the surrounding medium of specific mating-inducing factors (gamones) (Luporini, P et al., J exp zool 226 (1983) 1 [9]). The gamone from the mating type represented by strain 13 has been isolated and identified as a glycoprotein with a molecular weight (MW) of about 12 kD and a pI of 4. It has been termed euplomone r 13. At a concentration of 3 X 10(-12) M, euplomone r 13 specifically induces cells of a complementary mating type to unite in conjugation within 2 h.

On the role of germ cells in planarian regeneration. II. Cytophotometric analysis of the nuclear Feulgen-DNA content in cells of regenerated somatic tissues.

Previous findings by our group have shown how primordial male germ cells take part in regenerative blastema formation in planarians by migrating to the wound. The role of these cells in rebuilding transected tissues has been investigated in a population of Dugesia lugubris s.l. which is particularly suited for our purpose. In fact, these planarians provide a clear karyological marker to distinguish diploid male germ cells (2n = 8) from tryploid embryonic or somatic cells (3n = 12). In this study we employed the cytophotometric analysis of the nuclear Feulgen-DNA content in order to distinguish non-replicating male germ cells from reserve and somatic cells. The Feulgen-DNA content in cells from the gonad-free caudal area was measured after complete regeneration. Most non-replicating cells (94-95%) were found to have a DNA amount typical of cells previously estimated as triploid. Some (5-6%) nuclei containing a DNA amount typical of cells previously estimated as diploid male gonia were also found. These findings seem to support the view that primordial male germ cells also participate in rebuilding somatic tissues according to the field influence they encounter during regeneration. The possibility that metaplasia (or cell transdifferentiation) may occur in planarians is finally discussed.

Autoplastic vein grafts in experimental microsurgery.

The writers discuss the histological results of a study of twelve auto-plastic vein grafts performed by a microsurgical technique on the femoral artery of the rabbit. Our observations, which were made at periods of from one to twelve weeks after the grafts had been performed, revealed the details of the nature and chronological development of the structural changes that occurred in the wall of the graft.