Search

Data regarding antibody responses to severe acute respiratory syndrome coronavirus‐2 (SARS‐CoV‐2) in patients infected with COVID‐19 are not yet available. In this study, we aimed to evaluate serum antibody responses in patients regardless of the outcome. We measured the circulating immunoglobulin G (IgG) antibody levels in 60 subjects with a certified history of SARS‐CoV‐2 infection by using immunoenzymatic, chemiluminescent, and Neutralization assays. Half patients had a severe infection, the other half were pauci‐symptomatic. We analyzed their antibody response to see the trend of the humoral response. Our results showed a significant difference in circulating IgG level among the two groups. The neutralizing antibody response against SARS‐CoV‐2 was significantly higher among those who had severe disease. Furthermore, ten subjects from each group were screened twice, and a declining antibody trend was observed in pauci‐symptomatic individuals. These findings provide evidence that humoral immunity against SARS‐CoV‐2 in pauci‐symptomatic people is weak and may not be long‐lasting. This may have implications for immunity strategy and prevention, since it is still not clear whether a time‐dependent decrease of both circulating and neutralizing antibodies to nonprotective levels could occur in a longer time span and whether potential vaccines are able to induce a herd immunity and a durable response.

Human coronaviruses are known for their tropism for central nervous system and to be associated with neurological complications. SARS-CoV-2 pandemy represent a major health issue and if respiratory symptoms are at the forefront, neurological symptoms should be expected. Here we report the case of two patient that developed encephalitic symptoms with neuropsychological impairment and pathologic cerebrospinal fluid features concomitantly to SARS-CoV-2 documented infection. Both patient recovered promptly without treatment. This report raises the question of central nervous system involvement in SARS-CoV-2 infection and the need for investigation of neuropsychological complications in infected patient.

The majority of bushcricket species (Insecta: Orthoptera: Tettigoniidae) use acoustic signals for mate attraction. Song production and the sound-producing structures, therefore, can be argued to be under sexual selection. The sound-producing structures might determine differences in the acoustic parameters of male songs, like the fundamental frequency. We investigated the morphology and scaling relationships of the sound production organs in males of the bushcricket Poecilimon ampliatus. Selection acting on morphological traits can be inferred from allometric relationships. Our results show that heavier and larger males have bigger sound-producing organs: wing area scales positively with body mass as well as with body size. The length of the stridulatory file and the stridulatory teeth length both increase with wing area. In contrast, the number and density of teeth on the stridulatory file decrease in heavier males. As a result, males with larger wings and longer stridulatory files have a reduced teeth density. Positive isometric scaling was detected for the length of stridulatory teeth, as well as negative isometry for teeth number and teeth density. Wing area in turn seems to be under positive sexual selection, as the relationship of wing size on male body size show a steep positive slope (β = 2.4). The morphological results indicate positive selection on sound-producing structures and are in line with the female preference for heavier males in mate choice experiments.

The frequent development of drug resistance to targeted therapies in cancer patients has stimulated interest in strategies counteracting resistance. Combining immunotherapies with targeted therapies is one such strategy. In this context, we asked whether human NK cells can target melanoma cells that have acquired resistance to selective inhibitors targeting activating mutants of the B-Raf kinase (BRAF inhibitors, BRAFi). We generated drug-resistant cell variants in vitro from human BRAF-mutant melanoma cell lines MEL-HO, COLO-38, SK-MEL-37, 1520 and from primary melanoma cells freshly isolated from two patients. All drug-resistant cell variants remained susceptible to lysis by IL-2-activated NK cells; and two BRAFi-resistant lines (BRAFi-R) became significantly more susceptible to NK-cell lysis than their parental lines. This was associated with significant HLA class I antigen downregulation and PD-L1 upregulation on the drug-resistant lines. Although blocking HLA class I enhanced the extent of lysis of both BRAFi-R and parental cells to NK-cell-mediated lysis, antibody-mediated inhibition of PD1-PD-L1 interactions had no detectable effect. HLA class I antigen expression on BRAFi-R melanoma variants thus appears to play a major role in their susceptibility to NK-cell cytotoxicity. These findings suggest that NK-cell-based immunotherapy may be a viable approach to treat melanoma patients with acquired resistance to BRAF inhibitors.

Background and Objective Since the 1960s, lasers have been used in neurosurgery for surgical removal of intracranial tumors. Because of its limited penetration (2 mm) through tissues and its wavelength, which is useful in water medium, the 2-µ thulium laser has been applied primarily in urology. Its features are attractive for application under microscope magnification during neurosurgical procedures. The aim of this study was to evaluate the usefulness of the 2-µ thulium laser during microsurgical removal of intracranial meningiomas. Materials and Methods Twenty patients with a diagnosis of intracranial meningiomas were treated with surgical intervention using a 2-µ thulium laser together with bipolar forceps, cavitron ultrasonic surgical aspirator (CUSA) and traditional microdissection instruments. Surgical removal was divided in four phases: (1) dissection from the external structures; (2) coagulation and debulking; (3) dissection from the deep structures; and (4) coagulation and removal of the basal implant. During all these steps, we evaluated the percentage of usage of the 2-µ thulium laser comparing them with bipolar forceps and ultrasonic aspirator and blunt dissection. Results Thulium laser was used mainly during phases 2 and 4 for 43% and 48.7% of the total removal, respectively. Although also useful during phases 1 and 3, it was only used for 2.2% and 31.3%, respectively: traditional dissection with scissors and forceps was preferred. Conclusions Thulium laser seems to be a useful aid in the surgery of intracranial meningiomas, especially to debulk, shrink, and coagulate the mass and the basal implant. Lasers Surg. Med. 45: 148–154, 2013. © 2013 Wiley Periodicals, Inc.

In temperate regions, latewood is produced when cambial activity declines with the approach of autumnal dormancy. The understanding of the temporal (cambium activity vs dormancy) and spatial (phloem, cambial region, maturing xylem) regulation of key genes involved in the phenylpropanoid pathway during latewood formation represents a crucial step towards providing new insights into the molecular basis of xylogenesis. In this study, the temporal pattern of transcript accumulation of 12 phenylpropanoid genes (PAL1, C4H3/5, C4H4, 4CL3, 4CL4, HCT1, C3H3, CCoAOMT1, COMT2, COMT5, CCR2) was analyzed in maturing xylem and phloem of Picea abies during latewood formation. Quantitative reverse transcription-polymerase chain reaction analyses revealed a well-defined RNA accumulation pattern of genes involved in the phenylpropanoid pathway during latewood formation. Differences in the RNA accumulation patterns were detected between the different tissue types analyzed. The results obtained here demonstrated that the molecular processes involved in monolignol biosynthesis are not restricted to the cambial activity timeframe but continued after the end of cambium cell proliferation. Furthermore, since it has been shown that lignification of maturing xylem takes place in late autumn, we argue on the basis of our data that phloem could play a key role in the monolignol biosynthesis process.

The oncoprotein encoded by the HER-2 oncogene is a member of the HER family of receptor tyrosine kinases and is actually the first successfully exploited target molecule in new biomolecular therapies of solid tumors. The association of HER-2 overexpression with human tumors, its extracellular accessibility, as well as its involvement in tumor aggressiveness are all factors that make this receptor an appropriate target for tumor-specific therapy. In addition, HER-2 overexpression fosters its immunogenicity, as shown by the frequency of B and T cell-mediated responses against this oncoprotein in cancer patients, and it is being investigated as a promising molecule for either passive and active immunotherapy strategies. This review summarizes a number of immune intervention approaches that target HER-2 in breast cancer.

Facial hemangioma is usually isolated but its association with craniocervical arterial anomalies and structural brain malformations is well known. The acronym PHACE syndrome (posterior fossa malformation, facial hemangiomas, arterial anomalies, cardiac/aortic anomalies, and eye abnormalities) has been used to indicate that disorder in which brain anomalies are mainly represented by the Dandy-Walker malformation. We report on a 10-month-old boy affected by facial hemangioma and a complex cortical dysplasia located in the left frontal region. The lesion was characterized by a deeply infolding pachygyric cortex and a band of gray matter lining the wall of the lateral ventricle. The entire left cerebral hemisphere appeared hypoplastic. No anomalies of the posterior fossa structures or cardiac/aortic malformations were present. An overlapping clinical/pathological pattern was previously reported in another patient with facial hemangioma and cerebrovascular anomalies. These observations seem to indicate that the facial hemangiomas may be associated with disorders of the cortical development.

Melanoma-associated peptides recognized by cytolytic T lymphocytes (CTL) in the context of several histocompatibility leukocyte antigens (HLA) are required for the development of specific immunotherapies. Using a transient transfection assay into COS-7 cells, we identified the gp100/pMel17 melanosomal protein as the shared antigen recognized by three independent CD8+ CTL clones in HLA-A*6801-restricted fashion. This finding was confirmed by the correlation between lack of gp100/pMel17 protein in a number of HLA-A*6801-positive melanomas and their resistance to lysis/cytokine production by the specific effectors. The gp100/pMel17 antigenic epitope was identified based on recognition of subfragments and on a computer-based prediction algorithm. Among a panel of gp100/pMel17-derived synthetic peptides only the 10-mer HTMEVTVYHR (gp100/pMel17182-191) induced tumor necrosis factor (TNF) release by CTL clones when pulsed on suitable target cells whereas both the 10-mer and the shorter 9-mer gp100/pMel17183-191 sensitized the same antigen-pulsed cells to lysis. In conclusion, the identification of the HTMEVTVYHR peptide will extend to HLA-A*6801 melanoma patients the possibility to exploit gp100/pMel17 melanosomal protein for experimental and clinical studies.

An efficient synthesis of the cyclic decapeptide MEN 11270 [H-DArg1-Arg2 Pro3-Hyp4-Gly5-Thi6-Dab7-DTic8-Oic9-Arg10 c(7gamma - 10alpha)] was developed. Two three-dimensional orthogonal strategies were applied and compared: Fmoc/Tos/Boc (procedure A) and Fmoc/Pmc/Dde (procedure B). Both resulted in a 23-step strategy comprising the stepwise solid-phase chain assembly of the linear protected peptide, partial deprotection, solution-phase cyclization and final full deprotection. The stepwise assembly of the linear peptide was optimized by double coupling and acylation time prolongation for critical residues (Tic, Dab, Thi, Pro). O-(7-azabenzotriazol-1-yl)-N,N,N',N' tetramethyluronium (HATU) was preferred as coupling reagent for Dab. In the cyclization step, the partial racemization of Arg10 (31% using 1-ethyl-3-(3'-dimethyl-aminopropyl) carbodiimide/1-hydroxybenzotriazole (EDC/HOBt) as activation system) was reduced to 3% with HATU. The final deprotection was performed in the presence of dimethylsulfide (procedure A) and thiocresol (procedure B) as scavengers, to avoid the sulfation of Hyp side chain. The final compound and the main by-products were characterized by mass spectroscopy (MS), nuclear magnetic resonance (NMR) and racemization test. Procedure B produced operationally simpler and more efficient results than A (28% overall yield versus 4%).

Aims The purpose of this study was to establish whether the cell proliferation index assessed by the monoclonal antibody MIB-1 would correlate with survival in pleural malignant mesothelioma. Methods and results We studied a series of seven long-term survivors with pleural malignant mesothelioma and a group of control cases with short-term survival. All cases showed MIB-1 positive cells, and labelling indices were expressed as percentage of cells with positive nuclear immunostaining by randomly counting 1000 tumour cells. A statistically significant difference was found between MIB-1 values in the long-term survival group and the control cases with short-term survival. Conclusions Our results indicate that the differences in biological behaviour of malignant mesothelioma in long-term and short-term survivors may be explained in part by differences in tumour growth fraction and that proliferation index could represent an important prognostic parameter for this tumour.

Background As diabetes mellitus represents a situation in which production of peroxides is increased, the aim of this study was to investigate the relationship between plasma and platelet levels of ascorbic acid (AA)/dehydroascorbic acid (DHA) and those of malonyldialdehyde (MDA), an indirect marker of lipoperoxides, both assayed using high-performance liquid chromatography (HPLC), in 59 patients with insulin-dependent diabetes mellitus (IDDM) compared with 51 healthy control subjects matched for sex, age, smoking habits, as well as for dietary intake of energy, alcohol and vitamin C. Results Mean plasma and platelet MDA were significantly higher in the patients affected with IDDM than in control subjects. Moreover, the diabetic group was characterized by a huge decrease in plasma AA [8.45 ± 5.5 μmol L−1 (SD) vs. 33.4 ± 7.6 μmol L−1, P = 0.0001], mirrored by a significant increase in plasma DHA (11.9 ± 3.9 μmol L−1 vs. 3.9 ± 2.5 μmol L−1, P = 0.0001). No detectable DHA was observed in the platelets from both diabetic and control subjects, whereas AA was significantly increased in platelets from diabetic patients compared with control subjects (42.6 ± 7.4 vs. 34.8 ± 5.1 nmol 10−9 platelets, P = 0.0001). Platelet AA in the diabetic group was significantly inversely correlated with glycated haemoglobin (r = −0.34; P = 0.04) and directly with plasma AA (r = 0.39; P = 0.02), the sum of plasma AA + DHA (r = 0.44; P = 0.009) and with platelet MDA (r = 0.38; P = 0.02). Conclusion (a) The ratio plasma AA/DHA is significantly lowered in IDDM in association with an increase in MDA levels; (b) only AA is detected in platelets, being augmented in the diabetic group; (c) plasma ascorbate depletion does not reflect platelet levels of AA; and, finally, (d) metabolic control, as well as intracellular lipoperoxides, modulates platelet AA in IDDM.

Background Both lipoprotein (a) [Lp(a)] and angiogenesis have been shown to be associated with initiation and progression of atherosclerotic plaque. Lp(a) and two neutralizing anti-Lp(a) antibodies were investigated for their capacity to affect the vasoproliferative processes of the chick embryo chorioallantoic membrane (CAM), a useful model for such an investigation. Methods Gelatin sponges loaded with Lp(a) alone or together with anti-Lp(a) antibodies, or with vehicle alone, phosphate-buffered saline (PBS), were implanted in vivo onto the CAM at incubation day 8. Four days later, sponges and the adjacent CAM tissues were assessed for the extent of angiogenesis in terms of microvessel counts. Results Lp(a)-loaded sponges gave significantly higher counts than those loaded with the Lp(a)–anti-Lp(a) antibodies complex, which overlapped those treated with PBS. The angiogenic response was similar to that obtained with basic fibroblast growth factor, a well-known angiogenic molecule. Conclusion These data suggest that Lp(a) is capable of inducing angiogenesis in vivo, which might account for its ability to enhance and support atherosclerosis.

Hexahydro-, 5b-1 and 6a,f,1 and tetrahydrophospholo[2,3-d]isoxazoles 8, 9 and 10 were synthesized by 1,3-dipolar cycloaddition of nitrones 3b-1 and benzonitrile oxide (4) to 2,3-dihydro-1-phenyl-1H-phosphole 1-oxide (1) and 2,3-dihydro-1-ethoxy-1H-phosphole 1-oxide (2). The structural assignment to the compounds was confirmed by an X-ray study of two compounds of the series 5a and 5m. The compounds show a good activity as fungicides against Plasmopara viticola on vines and against Botrytis cinerea on apples. Compounds 5a-d showed weak to moderate activity as herbicides.

Homeobox-containing genes comprise a gene family coding for transcription factors involved in normal development. Class I human homeobox (HOX) genes display a peculiar chromosomal organization, perhaps directly related to their function. Aberrant expression of homeobox genes has been associated with both morphological abnormalities and oncogenesis. We have reported that HOX gene expression is (i) specific for normal adult human organs (kidney, colon, lung) and (ii) altered in cancer specimens according to their histological type and stage of tumor progression. Here, we have investigated whether patterns of HOX gene expression are associated with tumor heterogeneity by analyzing the expression of the entire panel of 38 HOX genes in clones isolated from a single human metastatic melanoma call line (Me 665/2). The differential expression of a block of genes located at the 5' end of the HOX C locus allows melanoma clones to be classified into 2 major groups. The 2 patterns of HOX gene expression are inversely associated with 2 distinct surface phenotypes for integrins (VLA-2, VLA-5 and VLA-6) and the adhesion molecule ICAM-1. The genes of the HOX C locus are silent in the clones with high levels of integrins VLA-2, VLA-5 and VLA-6 and of the adhesion molecule ICAM-1 but actively expressed in the clones with low levels of ICAM-1 and lacking VLA-2, VLA-5 and VLA-6. Our results indicate that HOX gene expression reflects the intra-tumor heterogeneity of melanoma clones and suggest that the expression of surface molecules involved in cell-cell and cell-matrix interactions may be related to the patterns of HOX gene expression.

Reduced glutathione (GSH) and activity of GSH related enzymes play a key role in defence against oxygen free radicals, whose production is, as known, raised in patients affected by diabetes mellitus, and at the same time they may contribute to the process of platelet aggregation. The purpose of this study was to evaluate GSH levels and activity of glutathione peroxidase (GSH-Px), glutathione reductase (GSSG-Red), glutathione transferase (GSH-Tr), glucose-6-phosphate-dehydrogenase (G6PDH), and thioltransferase (TT) in platelets of insulin-dependent diabetic patients in fair metabolic control (mean glycated haemoglobin: 6.5%), as related to presence of retinopathy, neuropathy or nephropathy and to platelet aggregation by arachidonic acid (AA) in vitro. Mean effective dose (ED50) of AA was on average significantly lower in the group of insulin-dependent diabetic patients (0.41 +/- 0.02 mM (SEM), n = 46) as compared with that of control subjects strictly matched for age, sex and weight (0.77 +/- 0.02, n = 51; P = 0.0001). Mean platelet GSH as well as the activity of GSH related enzymes expressed as geometric mean (95% confidence intervals) were similar in diabetic patients and in controls, except for GSSG-Red whose activity was significantly higher in diabetic subjects (28.5 (14.4-57.5) mU 10(-9) platelets vs. 20.3 (8.7-56) mU 10(-9) platelets; P = 0.01). In the diabetic group TT was reduced when compared with healthy controls (3.8 (0.9-12.2) mU 10(-9) platelets vs. 6 (1.6-26.1) mU 10(-9) platelets; P = 0.04).(ABSTRACT TRUNCATED AT 250 WORDS)

The frequencies of cytotoxic T-lymphocyte precursors (CTLp) that lyse autologous tumor by a T-cell receptor (TCR)-dependent mechanism (specific CTLp) were evaluated by limiting dilution analysis (LDA) using lymphocytes from peripheral blood (PBL) and from surgically resected, tumor-invaded lymph nodes (LNL) in 9 melanoma patients. The frequency of specific CTLp was determined in PBLs and/or LNIs of all patients by a modified LDA assay, enabling us to measure lytic activity on the autologous tumor that could be significantly inhibited by an anti-CD3 monoclonal antibody (MAb). This assay allowed us to detect frequencies of specific CTLp ranging from 1/720 to 1/32,037 in peripheral blood and from 1/328 to 1/22,061 in tumor-invaded lymph nodes. These frequencies indicated that lymphoid populations from PBLs or LNLs of melanoma patients may contain as low as 30 to as much as 3,000 specific CTLp/10(6) lymphocytes. In addition, comparison of wells containing specific CTLp with those showing no inhibition by anti-CD3 MAb indicated that specific CTLp represent between 3 and 88% of all precursors with lytic activity on the tumor. In 6 of 9 patients, no marked differences between PBLs and LNIs in specific CTLp frequencies were found. A 10-fold increase of specific CTLp, in comparison to PBL and LNL, was found only in lymphocytes isolated from a subcutaneous metastasis of one patient. Our results indicate that CTLp interacting with autologous tumor by a TCR-dependent mechanism exist in PBL and LNL of most melanoma patients, although a wide variation in their absolute number is evident among different patients.

The cycloaddition of cyclic nitrones to methoxycarbonylmethylenecyclopropane ( 1 ) gives adducts having the methoxycarbonyl group on C-4 of the isoxazolidine ring with high regioselectivity. The thermal rearrangement of the adducts gives quinolizidinone 6 and indolizidinones 8 bearing the methoxycarbonyl group selectively at C-1 and at C-8, respectively. The two compounds are intermediates for new formal syntheses of the alkaloids (±)-Lupinine and (±)-Epilupinine, and (±)-Elaeokanine A, respectively.

We report here the molecular characterization of the T-cell receptor (TCR) expressed by a human HLA-A2 specific cytotoxic T-cell clone named CTL 49. Flow cytometry analysis with a panel of anti-TCR antibodies revealed an OKT3+, WT31+, A13+, BB3-, TCR delta-, delta TCS1-, TCR gamma/delta 1-, OKT4-, and OKT8+ phenotype, suggesting that, in CTL 49, the V delta 1-encoded A13 epitope could be included in its alpha beta TCR. Northern blot analysis confirmed the presence of C alpha, C beta and V delta 1 specific transcripts while no hybridization signal was detected by a C delta specific probe. Polymerase chain reaction (PCR) amplification of the first strand cDNA from CTL 49 with TCR-specific primers and sequence analysis revealed that V delta 1 region is productively rearranged to J alpha and to C alpha regions. This alpha chain pairs with a beta chain composed of V beta 13.2/D beta/J beta 2.3/C beta 2 leading to the expression of a functional TCR complex. These results, in addition to providing further evidence for the sharing of V delta 1 by alpha/beta and gamma/delta TCR, indicate that an alpha/beta T-cell receptor which includes the V delta 1 variable region can be involved in alloreactive recognition.

Expression of alpha and beta subunits of VLA and VNR integrins was analyzed by cytofluorimetric analysis on 6 different human primary and metastatic melanoma cell cultures. Marked inter-tumor heterogeneity was observed, and expression of VLA-alpha I, VLA-alpha 2 and VLA-alpha 6 was lower on primary melanomas than on metastatic lesions. The function of VLA products on melanoma cells was assessed by adhesion assays to extracellular matrix (ECM) proteins using a panel of melanoma clones previously characterized for the presence and heterogeneity of expression of the distinct VLA-alpha subunits. These experiments indicated that intra-tumor heterogeneity in the integrin profile can influence the interaction of neoplastic cells with ECM proteins. Inhibition of adhesion with antibodies to VLA-alpha subunits revealed that the presence on melanoma cells of VLA-alpha 2, VLA-alpha 5 and VLA-alpha 6 is relevant for the adhesion to type-IV collagen, fibronectin and laminin respectively. Culture of tumor cells in the presence of cytokines such as rIL-I beta, rTNF-alpha, rIFN-gamma or TGF-beta I could induce up- or down-modulation in the level of expression of multiple VLA integrins. Cytokine-mediated antigenic shifts in the VLA profile of melanoma cells were detected by cytofluorimetric analysis as early as 24 hr after cytokine exposure. The cytokine-dependent change in the matrix receptor profile of melanoma cells also affected the adhesion to ECM proteins as revealed by the enhanced adhesion of rTNF-alpha-treated cells to fibronectin. These data indicate that constitutive heterogeneity in the integrin profile or cytokine-mediated shifts in VLA expression can affect the ability of human melanoma cells to interact with different ECM components.