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Biomedical subjects

A Franchini

Publications and source records attributed to A Franchini.

At least 37 records · Page 2Linked to original sources

Morula cells as the major immunomodulatory hemocytes in ascidians: evidences from the colonial species Botryllus schlosseri.

Immunocytochemical methods were used to study the presence and distribution of IL-1-alpha- and TNF-alpha-like molecules in the hemocytes of the colonial ascidian Botryllus schlosseri. Only a few unstimulated hemocytes were positive to both the antibodies used. When the hemocytes were stimulated with either mannan or phorbol 12-mono-myristate, the phagocytes were not significantly changed in their number, staining intensity, or cell morphology. In contrast, stimulated morula cells were intensely labeled, indicating that these cells play an important immunomodulatory role.

Animals↗

Immunomodulation by recombinant human interleukin-8 and its signal transduction pathways in invertebrate hemocytes.

We report the presence of interleukin (IL)-8-immunoreactive molecules in hemocytes from the mollusc Mytilus galloprovincialis. Functional studies demonstrate that recombinant human (rh)IL-8 provokes conformational changes, induces chemotaxis, and increases bacterial phagocytic activity in hemocytes. rhIL-8 induces cell shape changes via protein kinase A and C pathways. These morphological changes are followed by reorganization of the actin microfilaments. The findings suggest that, as previously reported for other cytokines, IL-8 is well conserved and deeply involved in immune functions from invertebrates to mammals.

Animals↗

PDGF and TGF-beta partially prevent 2-deoxy-D-ribose-induced apoptosis in the fat body cell line IPLB-LdFB from the insect Lymantria dispar.

The IPLB-LdFB cell line from the fat body of the insect Lymantria dispar shows the presence of immunoreactive, platelet-derived growth factor (PDGF)-AB and transforming growth factor (TGF)-beta1 molecules, as well as the corresponding plasma membrane-like receptors, i.e. PDGFR-alpha, PDGFR-beta and TGFR-beta type II. Cytofluorimetric and morphological studies reveal that the reducing sugar 2-deoxy-D-ribose (dRib), an apoptotic agent for human cells, induces apoptosis in a concentration- and time-dependent manner even in IPLB-LdFB cells. PDGF-AB and TGF-beta1 partially counteract the effect of dRib, indicating a survival role of these factors in this apoptotic model of insect cells.

Journal Article↗

Synergistic role of cAMP and IP(3) in corticotropin-releasing hormone-induced cell shape changes in invertebrate immunocytes.

Immunocytes from the mollusc Mytilus galloprovincialis express corticotropin-releasing hormone (CRH) receptor subtype (CRH-R1 and CRH-R2)-like mRNAs. Using computer-assisted microscopic image analysis, we have found that exogenous CRH provokes changes in the cellular shape of immunocytes, and that this response is extracellular Ca(2+)-dependent. The various inhibitors of transduction signaling pathways, i.e. suramin sodium, 2', 5'-dideoxyadenosine, neomycin sulfate, calphostin C, H-89, and wortmannin, completely or partially inhibit these changes. The present findings demonstrate that PKA, PKC, and PKB/Akt are involved in CRH-induced cell shape changes in immunocytes, and that the cellular effect of CRH needs the synergistic action of the two second messengers, cAMP and IP(3).

Animals↗

Human placenta, chorion, amnion and decidua express different variants of corticotropin-releasing factor receptor messenger RNA.

Human placenta is a major source of corticotropin-releasing factor (CRF), and local effects of CRF in fetal membranes and placenta have been shown, i.e., adrenocorticotropic hormone (ACTH) and oxytocin release from cultured placental cells, as well as prostaglandin release from amnion, chorion and decidua. Two distinct CRF receptors (CRF-R1 and CRF-R2) have been characterized: CRF-R1 consists of two isoforms (CRF-R1alpha and CRF-R1beta) while CRF-R2 has at least three different splice variants (CRF-R2alpha, CRF-R2beta and CRF-R2gamma). To date, CRF-R1 receptor has been identified in human placenta and in pregnant myometrium, while no evidence for placental CRF-R2 receptor isoforms has been provided. The present study investigated whether the different isoforms of CRF-R1 and CRF-R2 receptor mRNA are expressed in fetal membranes and placenta. Tissues were collected after spontaneous vaginal delivery (38-40 weeks) or elective caesarean section (39-41 weeks). The gene expression of CRF receptors was first studied by reverse transcriptase-polymerase chain reaction (RT-PCR), and the presence of CRF-R1alpha, but not of CRF-R1beta, in human placental trophoblast, amnion/chorion and decidua was shown. In addition, among the three CRF-R2 splice variants, only CRF-R2beta mRNA was expressed by trophoblast and fetal membranes. By using in situ hybridization, CRF-R1 and CRF-R2 probes positively hybridized trophoblast and related membranes. CRF-R1 was localized in the syncytiotrophoblast cells, chorionic trophoblast and decidua with a small amount in the amnion. CRF-R2 probe mainly hybridized syncytiotrophoblast cells, but cytotrophoblast also contained discreet amounts of CRF-R2 mRNA signal. The CRF-R2 hybridization signal was also observed within the structure of the villi (blood vessels), chorionic trophoblast and decidual cells, but it was faint or absent in the amniotic epithelium. There was no significant difference in the distribution of CRF-R1 or CRF-R2 mRNA signal between placentas collected from vaginal delivery or caesarean section. The evidence that intrauterine tissues differently express CRF-R1alpha and CRF-R2beta supports possible different local roles of CRF and related peptides into intrauterine tissues during pregnancy.

Amnion↗

Repair of molluscan tissue injury: role of PDGF and TGF-beta1.

Various cellular and humoral activities of the wound repair process and the effects of PDGF-AB and TGF-beta1 on tissue repair mechanisms in the mollusc Limax maximus are studied by histological and immunocytochemical procedures. Histological examination at different times after the wound production demonstrates that tissue repair is the result of successive and related activities distinguishable by different morphological pictures. In the first hours, an infiltration phase is activated 24 h after the incision, hemocytes stratify at wound margins and actively phagocitize cell debris and damaged tissue in the surrounding area. Moreover, the cells are immunoreactive to anti-IL-1alpha, IL-8 and TNF-alpha antibodies. After 24-72 h, granulation tissue rich in small blood lacunae is formed and the provisional matrix is synthesized and deposited on the base of the new tissue. In histological sections 72 h after injury, the incision is filled with granulation tissue, and at the wound base, a layer of fibrous connective tissue is formed. Hemocytes present in the newly formed blood lacunae and fibroblasts are involved in the synthesis and deposit of extracellular matrix components, i.e. fibronectin, reticular and collagen fibres. Ninety-six h after wound production, the repair process continues and the granulation tissue is more developed. At 192 h, re-epithelialization begins, and this is more evident in the histological sections after 14 days. Hemocytes are immunoreactive to the cytokines at all the times examined. Exogenous administration of PDGF-AB and TGF-beta1 stimulates the tissue healing process through a general acceleration of the activities involved. A larger closing area of clumped hemocytes and a smaller damaged tissue area are observed 24 h after treatment of the wound. At 72 h, the granulation tissue is more developed and more extracellular matrix components are deposited than in the control incision. A larger number of cells express cytokine-like molecules, and re-epithelialization of the wound is accelerated, as 14 days after growth factor treatments almost all the wound area is covered by a layer of cubic epithelial cells, and the alcianophilic cell coat is restored. No differences in the responses of the two growth factors are observed.

Animals↗

Effects of dietary vitamin E on the quality of table eggs enriched with n-3 long-chain fatty acids.

Because of the proposed cardioprotective benefits of n-3 fatty acids and vitamin E, a trial was carried out to investigate the possibility of enriching eggs with n-3 fatty acid and vitamin E added to the hen's diet. One hundred ninety-two Hy-Line Brown hens, 39-wk-old, were divided into eight groups: four groups received the basal diet supplemented with 3% lard and four doses of dl-alpha-tocopheryl acetate (0, 50, 100, and 200 ppm), whereas the diets of the other groups were supplemented with 3% of fish oil and the same doses of vitamin E. The performances of the hens and egg weights were not affected either by the type of lipid supplement or by the vitamin level. The treatment with fish oil caused a dramatic increase (P < 0.01) of all n-3 fatty acids of the yolk, particularly EPA (19.53 vs. 0.74 mg/egg) and DHA (143.70 vs. 43.66 mg/egg), and an appreciable decrease of arachidonic acid (25.54 vs. 67.72 mg/egg). The different levels of dietary vitamin E slightly affected the fatty acid composition of the yolk. Yolk alpha-tocopherol increased linearly as dietary dl-alpha-tocopheryl acetate increased (P < 0.01) from the control level of 90.93 microg/g of yolk to 313.84 microg/g of yolk when 200 ppm were added to the hen diets. Twenty-eight days of storage at room temperature (20 to 25 C) did not alter the yolk fatty acid profile, and, moreover, the levels of vitamin E remained still very close to those observed in fresh egg.

Animal Nutritional Physiological Phenomena↗

Presence of IL-1- and TNF-like molecules in Galleria mellonella (Lepidoptera) haemocytes and in an insect cell line Fromestigmene acraea (Lepidoptera).

In this study the authors give immunocytochemical evidence for the presence of interleukin (IL)-1alpha- and tumour necrosis factor (TNF) alpha-like molecules in the haemocytes of last instar larvae from the greater wax moth Galleria mellonella. Similar results are demonstrated in a continuous haemocyte line (BTI-EA-1174-A) from the salt marsh caterpillar Estigmene acraea. In Galleria mellonella larvae granular cells show a strong positive reaction with both primary antibodies, whereas plasmatocytes are stained to a lesser extent. Cell line haemocytes also react positively with both antibodies. After activating the cells with lipopolysaccharide (LPS) staining of Estigmene acraea cells is decreased, whereas Galleria mellonella haemocytes show no visible reaction in comparison to non-activated cells.

Animals↗

Detection of casein fragments in an invertebrate and in a vertebrate using in situ hybridization.

Using a non-radioactive in situ hybridization procedure it has been demonstrated that both invertebrates such as the mollusc Mytilus galloprovincialis and lower vertebrates such as the fish Cyprinus carpio express bovine alphaS1- and kappa-casein homologous mRNAs. In particular, positive results were found in molluscan immunocytes, and in cells located in different fish tissues: intestine, endocrine pancreas and kidney. These findings suggest that the casein genes are highly conserved throughout evolution.

Animals↗

Gill endocrine cells in the goldfish Carassius carassius var. auratus and their impairment following experimental lead intoxication.

The presence of endocrine cells in the gills of the goldfish Carassius carassius and the effects of lead intoxication (5 mg l(-1)) on their relative abundance and secretory activity were studied. Endocrine cells synthesizing peptido-like (met and leu-enkephalin and neuropeptide Y) and serotonin-like substances were detected in gill filaments by immunocytochemical procedures. Decreased immunoreactivity for two enkephalins and neuropeptide Y was observed after 48 and 96 h lead exposure. In contrast, increased serotonin immunoreactivity was evident after a one-week treatment. A search for Ca++-binding proteins was also carried out by immunocytochemical methods. Calbindin D-28K and S-100-like molecules were detected in gill epithelium, and their expression appeared enhanced after lead exposure.

Animals↗

Immunoreactive POMC-derived peptides and cytokines in the chicken thymus and bursa of Fabricius microenvironments: age-related changes.

Changes from hatching to the involutive stage in the thymus and bursa of Fabricius of Gallus domesticus were studied. Pro-opiomelanocortin (POMC)-derived peptides and cytokines were also tested by immunocytochemical procedures. Thymic histological modifications appeared at 3 months and involved an increase in connectival argyrophilic reticular fibres, a proliferation of non-epithelial cells in the reticulum network, the presence of mucous cells and small mucous cysts, and an increase in the number of eosinophilic and myoid cells. A clear distinction between the cortex and the medulla was lost. Immunoreactive POMC-related molecules and cytokines were demonstrated in thymic cells from 4-day-old chicken, and their number increased with ageing. These molecules were expressed in a few single epithelial cells and in interdigitating cells. With ageing, the number of immunoreactive interdigitating cells also increased, and these appeared in an activated phase. Histological modifications in the bursa of Fabricius appeared at 2 months and concerned a folding of the interfollicular surface epithelium covering the bursal plicae and a reduction in lymphoid follicle-associated epithelium. Fibrous tissue gradually increased, and large mucoid cysts were evident. The expression of POMC-derived peptide-and cytokine-like molecules differed during the development and involution phases of the organ. Cells of follicle associated epithelium and dendritic reticular cells of lymphoid follicles were immunoreactive to beta-endorphin between 4 days and 2 months, while ACTH-, alpha-MSH- and cytokine-like molecules were observed in follicles after 2 months. The findings indicate a physiological role of these molecules during the growth and involution of the two organs.

Aging↗

Automated genome sequence analysis and annotation.

MOTIVATION: Large-scale genome projects generate a rapidly increasing number of sequences, most of them biochemically uncharacterized. Research in bioinformatics contributes to the development of methods for the computational characterization of these sequences. However, the installation and application of these methods require experience and are time consuming. RESULTS: We present here an automatic system for preliminary functional annotation of protein sequences that has been applied to the analysis of sets of sequences from complete genomes, both to refine overall performance and to make new discoveries comparable to those made by human experts. The GeneQuiz system includes a Web-based browser that allows examination of the evidence leading to an automatic annotation and offers additional information, views of the results, and links to biological databases that complement the automatic analysis. System structure and operating principles concerning the use of multiple sequence databases, underlying sequence analysis tools, lexical analyses of database annotations and decision criteria for functional assignments are detailed. The system makes automatic quality assessments of results based on prior experience with the underlying sequence analysis tools; overall error rates in functional assignment are estimated at 2.5-5% for cases annotated with highest reliability ('clear' cases). Sources of over-interpretation of results are discussed with proposals for improvement. A conservative definition for reporting 'new findings' that takes account of database maturity is presented along with examples of possible kinds of discoveries (new function, family and superfamily) made by the system. System performance in relation to sequence database coverage, database dynamics and database search methods is analysed, demonstrating the inherent advantages of an integrated automatic approach using multiple databases and search methods applied in an objective and repeatable manner. AVAILABILITY: The GeneQuiz system is publicly available for analysis of protein sequences through a Web server at http://www.sander.ebi.ac. uk/gqsrv/submit

Amino Acid Sequence↗

ACTH and its role in immune-neuroendocrine functions. A comparative study.

Adrenocorticotropic hormone (ACTH) belongs to the melanocortine group of related peptides which share a common precursor, the pro-opiomelanocortin (POMC). Melanocortin expresses its functional effects by the stimulation of specific G-protein coupled receptors. To date, five receptor subtypes have been cloned. The POMC gene has been highly conserved during evolution, and ACTH has been found in different cells from invertebrates to vertebrates to vertebrates, including man. With regards the immune system, the presence of melanocortin receptors and the production of ACTH have been reported in invertebrate and vertebrate immuno-competent cells. Among its various physiological effects, ATCH has been shown to play a central role in immmune responses, such as chemotaxis and phagocytosis, in lower and higher forms of life. Moreover, ACTH is a key actor in stress response, and the complex cascade of events observed in vertebrates is reproduced and concentrated in intvertebrate immunocytes. On the basis of the present findings, ACTH should be considered an important immuno-regulator, forming part of the complex mosaic of relationships between the immune and neuroendocrine system which appears to have been substantially maintained over the course of evolution.

Adrenocorticotropic Hormone↗

Dynamics of hypervariable region 1 variation in hepatitis C virus infection and correlation with clinical and virological features of liver disease.

Hepatitis C virus (HCV) infection is a dynamic process during which molecular variants are continuously selected as the result of virus adaptation to the host. Understanding the nature of HCV genetic variation is central to current theories of pathogenesis and immune response. We prospectively studied hypervariable region 1 (HVR1) variation in the E2 gene of 36 hepatitis C patients, including 10 asymptomatic carriers, followed up for 1 to 2 years. Sequence changes in single and consecutive serum samples were assessed and correlated with clinical and virological parameters of liver disease. A region of the E1 gene was sequenced for comparison in 3 subjects. HVR1 heterogeneity at single time points widely varied in individual patients, did not increase cumulatively over the follow-up period, and did not correlate with HVR1 evolutionary rates. Conversely, the process of HVR1 sequence diversification, although differed considerably among patients, was stable over time and directly correlated with infections by HCV type 2, lower alanine aminotransferase (ALT) levels, and absence of cirrhosis. HCV carriers showed the highest HVR1 variation rates. Our findings indicate that HVR1 variation has an adaptive significance and is associated with favorable features of liver disease and suggest that prospective, rather than static, observations are required to model the process of HCV variation.

Adult↗

In situ localization of ACTH receptor-like mRNA in molluscan and human immunocytes.

Adrenocorticotropin hormone (ACTH) receptor-like messenger RNA was localized in molluscan hemocytes and human peripheral blood mononuclear cells by in situ hybridization using a digoxigenin-labelled bovine complementary DNA probe. These findings suggest that the ACTH receptor gene has been highly conserved during evolution. Moreover, these data represent further support for a relationship between the immune and neuroendocrine systems in invertebrates, as documented in our previous studies.

Animals↗

PDGF and TGF-beta induce cell shape changes in invertebrate immunocytes via specific cell surface receptors.

The presence of PDGF receptor-alpha- and -beta- and TGF-beta-receptor (type II)-like molecules on the plasma membranes of the immunocytes of the mollusc Mytilus galloprovincialis was demonstrated by an immunocytochemical procedure. Furthermore, the present study provides evidence that PDGF-AB and TGF-beta1 provoke cell shape changes in immunocytes via interactions with the respective receptors and that these extracellular signals are transduced along the phosphoinositide signaling pathway.

Animals↗

Glycoconjugate and adhesion molecule changes during the cell cycle in a human embryonic epithelial cell line.

The changes in the expression of glycoconjugates and adhesion molecules were studied by selective lectin binding and immunocytochemical reactions in a human embryonic epithelial cell line (EUE cells), synchronized in the cell phases. The results can be summarized as follows: most of the tested lectins display a more diffuse binding for the cytoplasm in G1 than S and G2 phases; in the S and particularly in G2 phases the cytoplasm glycoconjugates are arranged around the nucleus; cells in mitosis always show a strong binding towards all tested lectins. Cellular fibronectin and its receptor beta 1 integrin are well expressed in G1, but the strongest reaction is observed in the S phase. The immunoreactions for laminin and uvomorulin (L-CAM) are poorly positive in all cell cycle phases. The immunocytochemical reaction for heparan sulfate is positive, with a stronger reaction in S and G2 than G1; on the contrary a diffuse staining with the anti-dermatan sulfate proteoglycan antibody appears unchanged during the cell cycle.

Cell Adhesion↗