From the Bay of Naples to the River Don: the Campanian Ignimbrite eruption and the Middle to Upper Paleolithic transition in Eastern Europe

The Campanian Ignimbrite (CI) eruption, dated by ⁴⁰Ar/³⁹Ar and various stratigraphic methods to ca. 39,000 cal BP, generated a massive ash plume from its source in southern Italy across Southeastern and Eastern Europe. At the Kostenki-Borshchevo open-air sites on the Middle Don River in Russia, Upper Paleolithic artifact assemblages are buried below, within, and above the CI tephra (which is redeposited by slope action at most sites) on the second terrace. Luminescence and radiocarbon dating, paleomagnetism, and soil and pollen stratigraphy provide further basis for correlation with the Greenland and North Atlantic climate stratigraphy. The oldest Upper Paleolithic occupation layers at Kostenki-Borshchevo may be broadly correlated with warm intervals that preceded the CI event and Heinrich Event 4 (HE4; Greenland Interstadial: GI 12-GI 9) dating to ca. 45,000-41,000 cal BP. These layers contain an industry not currently recognized in other parts of Europe. Early Upper Paleolithic layers above the CI tephra are correlated with HE4 and warm intervals that occurred during 38,000-30,000 cal BP (GI 8-GI 5), and include an assemblage that is assigned to the Aurigancian industry, associated with skeletal remains of modern humans.     

Mesoscopic gel at low agarose concentration in water: a dynamic light scattering study.

Previous work in our laboratory has shown that at very low agarose concentration in water gelation still occurs within mutually disconnected, high concentration regions generated by spinodal demixing. The freely diffusing particles obtained in these conditions are studied in the present work by depolarized dynamic light scattering and probe diffusion experiments. These particles are found to behave as large (in fact, mesoscopic) polymer fibers entangled in a continuously rearranged mesh with scaling parameters typical of partially flexible, neutral chains. The present results allow specifying the notion of mesoscopic gelation. They also reveal that the same symmetry-breaking mechanism that allows macroscopic gelation at polymer concentrations well below the threshold for random cross-link percolation generates additional and unexpected phenomena.     

Elevated blood Hsp60, its structural similarities and cross-reactivity with thyroid molecules,and its presence on the plasma membrane of oncocytes point to the chaperonin as an immunopathogenic factor in Hashimoto's thyroiditis

The role Hsp60 might play in various inflammatory and autoimmune diseases is under investigation, but little information exists pertaining to Hashimoto’s thyroiditis (HT). With the aim to fill this gap, in the present work, we directed our attention to Hsp60 participation in HT pathogenesis. We found Hsp60 levels increased in the blood of HT patients compared to controls. The chaperonin was immunolocalized in thyroid tissue specimens from patients with HT, both in thyrocytes and oncocytes (Hurthle cells) with higher levels compared to controls (goiter). In oncocytes, we found Hsp60 not only in the cytoplasm but also on the plasma membrane, as shown by double immunofluorescence performed on fine needle aspiration cytology. By bioinformatics, we found regions in the Hsp60 molecule with remarkable structural similarity with the thyroglobulin (TG) and thyroid peroxidase (TPO) molecules, which supports the notion that autoantibodies against TG and TPO are likely to recognize Hsp60 on the plasma membrane of oncocytes. This was also supported by data obtained by ELISA, showing that anti-TG and anti-TPO antibodies cross-react with human recombinant Hsp60. Antibody-antigen (Hsp60) reaction on the cell surface could very well mediate thyroid cell damage and destruction, perpetuating inflammation. Experiments with recombinant Hsp60 did not show stimulation of cytokine production by peripheral blood mononuclear cells from HT patients. All together, these results led us to hypothesize that Hsp60 may be an active player in HT pathogenesis via an antibody-mediated immune mechanism.     

Haying and grazing effects on the butterfly communities of two Mediterranean-area grasslands

Grasslands are usually the most suitable environment for butterflies, but have been also traditionally used for productive activities. This paper compares the impact of mowing and grazing on butterfly biodiversity in two S Italian (Campania) grasslands, at Campo Somma (CS) and Pianoro di Prada (PP) located at an identical altitude of 850 m. These grasslands have an area of approximately 6 ha each and are at a 3 km distance from each other. They share similar climate and are both surrounded by woods, mainly of sweet chestnut. CS is managed for haying and is mown once a year, in June. PP is used for sheep and cattle grazing. Weekly transects were made from April to September in 2008 and 2009, during the butterfly flight activity. We analysed data from eight monthly transects by Kruskal–Wallis and Mann–Whitney tests. We observed 45 butterfly species at PP, and 28 at CS. Diversity indexes were significantly higher at PP, whereas evenness was similar. The monthly comparisons of species richness showed that, except for April, PP values were always significantly higher. Species abundance was significantly higher at PP in June, July and August. In all months, except in April, the Shannon–Wiener index was also significantly higher at PP, as was Simpson’s index in May, June and September. Dominance index differed significantly only in May, June and September, whereas evenness was never significantly different between the two grasslands. These data show that, as concerns butterflies, the impact of mowing was much stronger than that of grazing.     

N-Glycans mutations rule oligomeric assembly and functional expression of P2X3 receptor for extracellular ATP

N-Glycosylation affects the function of ion channels at the level of multisubunit assembly, protein trafficking, ligand binding and channel opening. Like the majority of membrane proteins, ionotropic P2X receptors for extracellular ATP are glycosylated in their extracellular moiety. Here, we used site-directed mutagenesis to the four predicted N-glycosylation sites of P2X(3) receptor (Asn(139), Asn(170), Asn(194) and Asn(290)) and performed comparative analysis of the role of N-glycans on protein stability, plasma membrane delivery, trimer formation and inward currents. We have found that in transiently transfected HEK293 cells, Asn(170) is apparently the most important site for receptor stability, since its mutation causes a primary loss in protein content and indirect failure in membrane expression, oligomeric association and inward current responses. Even stronger effects are obtained when mutating Thr(172) in the same glycosylation consensus. Asn(194) and Asn(290) are the most dispensable, since even their simultaneous mutation does not affect any tested receptor feature. All double mutants containing Asn(170) mutation or the Asn(139)/Asn(290) double mutant are instead almost unable to assemble into a functional trimeric structure. The main emerging finding is that the inability to assemble into trimers might account for the impaired function in P2X(3) mutants where residue Asn(170) is replaced. These results improve our knowledge about the role of N-glycosylation in proper folding and oligomeric association of P2X(3) receptor.     

The Di Bella Method (DBM) improved survival, objective response and performance status in a retrospective observational clinical study on 23 tumours of the head and neck

In 23 cases of carcinoma of the head and neck, the combined use of Somatostatin and/or its analogue Octreotide, prolactin inhibitors, Melatonin, Retinoids, Vitamin E, Vitamin D3, Vitamin C, Calcium, chondroitin-sulphate, and minimal oral doses of cyclophosphamide (50-100 mg/day) led to a decided increase in survival with respect to the median values reported in the literature for the same tumours and stages, together with an evident improvement in the quality of life, partial or complete objective responses and, in some cases, complete and stable cure with functional recovery. The rationale and the mechanisms of molecular biology of the treatment are discussed, showing that the treatment has a differentiating, apoptotic, antiproliferative, antiangiogenic and antimetastatic effect, and, unlike chemo- and/or radiotherapy, preserves and enhances the trophism and functionality of organs, tissues and immunitary and antitumoral homeostasis. This result, achieved without toxicity, demonstrates the efficacy of this biological multitherapy (Prof. Luigi Di Bella's method or DBM) and is in agreement with the positive results already published on the use of the DBM in various neoplastic diseases. We believe it is of use to report these cases to invite greater interest in the possibilities opened up by this biological multitherapy.     

Quantitative analysis of the impact of a human pathogenic mutation on the CCT5 chaperonin subunit using a proxy archaeal ortholog

The human chaperonin complex is a ~ 1 MDa nanomachine composed of two octameric rings formed from eight similar but non-identical subunits called CCT. Here, we are elucidating the mechanism of a heritable CCT5 subunit mutation that causes profound neuropathy in humans. In previous work, we introduced an equivalent mutation in an archaeal chaperonin that assembles into two octameric rings like in humans but in which all subunits are identical. We reported that the hexadecamer formed by the mutant subunit is unstable with impaired chaperoning functions. This study quantifies the loss of structural stability in the hexadecamer due to the pathogenic mutation, using differential scanning calorimetry (DSC) and isothermal titration calorimetry (ITC). The disassembly of the wild type complex, which is tightly coupled with subunit denaturation, was decoupled by the mutation without affecting the stability of individual subunits. Our results verify the effectiveness of the homo-hexadecameric archaeal chaperonin as a proxy to assess the impact of subtle defects in heterologous systems with mutations in a single subunit.