Amygdala structure and function and its associations with social-emotional outcomes in a low-risk preterm sample.

Amygdala atypical volume development and functional connectivity (FC) at small gestational ages (GA) have been found across childhood. This adult-oriented study assesses whether altered amygdala structure and function is present following low-risk preterm birth. T1-weighted and resting-state functional MRI images of 33 low-risk preterm (30-36 weeks' GA) and 29 full-term (37-42 weeks' GA) young adults of both sexes, aged between 20 and 32 years old, were analyzed using FreeSurfer (v6.0.0) and Coon Toolbox (v21.a). The social-emotional assessment included Happé's Strange Stories Test, the Moral Judgment Test, Delay-Discounting Test, Adult Self Report, and Emotion Regulation Questionnaire. No differences were found in social-emotional outcomes or amygdala volumes between the groups. Low-risk preterm young adults showed increased FC between the left amygdala, right amygdala and medial frontal cortex (MedFC) (F = 9.89, p-FWE = 0.009) at cluster level compared to their full-term peers. However, significant results at connection level were not observed between left and right amygdala. Lastly, increased FC at cluster level between the right amygdala and MedFC, and left amygdala and MedFC, was related to better social-emotional outcomes only in low-risk preterm young adults (F = 6.60, p-FWE = 0.036) at cluster level. At connection level, in contrast, only right amygdala-MedFC increased FC was significantly associated with better social-emotional outcomes. This study reveals that low-risk prematurity does not have an effect on social-emotional outcomes or structural amygdala volumes during young adulthood. However, individuals who were considered to be at a lower risk of exhibiting neurodevelopmental alterations following preterm birth demonstrated increased FC between the left and right amygdala and MedFC.

Absence of a relationship between the increased survival of skin grafts provoked by additional spleen transplantation and the detection of donor chimeric cells in rats.

The goal of this study was to examine the relationship between the effects of spleen transplantation on the acceptance of a skin allograft from the same donor and the detection of donor chimerism in recipient tissues. Male Sprague-Dawley rats and female Wistar rats were used as donors and recipients, respectively. Four experimental groups were established: group C: only skin grafting was performed; group S: recipients were splenectomized before skin grafting; group 1: skin grafts were performed immediately after splenectomy of recipients and spleen transplantation, and group 2: splenectomy of the recipients and spleen grafting were performed 48 h before skin grafting. All animals were sacrificed 14 weeks after surgery. The rate of acceptance of skin grafts was significantly higher in animals that received both skin and spleen transplantations than in group C (p = 0.03) or in group S (p = 0.04). Detection of donor chimeric cells was significantly more frequent in rats after spleen transplantation than in group C (p = 0.03). However, the detection of chimeric cells was not related to the acceptance of skin grafts. To conclude, the beneficial effect of spleen auxiliary transplantation on allograft survival was not related to the detection of long-term chimerism in the recipient's tissues.

[Low grade astroblastoma: pathological and magnetic resonance findings]. / Astroblastoma de baixo grau: Achados patológicos e da ressonância magnética.

INTRODUCTION: The astroblastoma is an uncommon type of glial tumour. It accounts for less than 1% of all tumours of the central nervous system. It originates in the tanicytes, ependymal cells present in the embryo and usually seen in adolescents and young adults. Radiologically it presents as a well delimited, heterogeneous tumour with a solid component which takes up contrast and is cystic, giving the same signal as cerebrospinal fluid (CSF). The pathological characteristics are of the formation of radial astroblastic pseudorosettes with perivascular hyalinization. CASE REPORT: An 18 year old woman presented with a 15 month history of motor deficit of her right limbs with occasional left frontal headache and horizontal diplopia on looking towards the left. On examination there was minimal claudication of the right limbs and bilateral papilloedema. Magnetic resonance showed a very well circumscribed left prefrontal neoplasm of heterogeneous aspect, with areas of solid and cystic appearance in the different sequences. The area of cystic appearance did not show the CSF signal in all sequences of the pulse. The tumour was totally excised. On histological study there were radial astroblastic pseudorosettes with perivascular hyalinization, with two mitoses per 10 fields of great magnification and the final diagnosis was of low grade astroblastoma. The apparently cystic portion was composed of friable gelatinous tissue. Fifteen months after her operation the patient is still asymptomatic. CONCLUSION: We report the radiological and pathological findings of a low grade astroblastoma.

Experimental evidence does not support use of the "no-touch" isolation technique in colorectal cancer.

PURPOSE: The benefits of the "no-touch" isolation technique usually performed to prevent the circulation of tumor cells are not evident. The aim of this study was to determine whether conventional surgical procedures for treatment of colon cancer could provoke the circulation of tumor cells detected by a genetic technology. METHODS: Sixteen patients undergoing resection for colorectal cancer and two patients with irresectable tumors were studied. No patient showed liver or lung metastasis. With specific primers for carcinoembryonic antigen, we used reverse transcriptase-polymerase chain reaction to analyze tumor biopsy specimens and blood samples obtained from the antecubital vein before and after surgery and from the main drainage vein of the tumor when the tumor had been extracted. Peritoneal fluid was also collected in irrecsectable cases. RESULTS: Amplification of cDNA with carcinoembryonic antigen-specific primers was achieved with all tumor biopsies and samples of peritoneal fluid. In two patients carcinoembryonic antigen reverse transcriptase-polymerase chain reaction products were detected in antecubital vein blood before surgery and in one of them also after surgery. Only in one patient (Dukes C) were carcinoembryonic antigen reverse transcriptase-polymerase chain reaction products detected from the main drainage vein of the tumor. In serial dilution experiments we determined that the limit of detection of this method was ten tumor cells in 2 ml of blood. CONCLUSION: Our data suggest that the use of no-touch isolation techniques in colorectal cancer is not justified, based on lack of evidence indicating the detachment of cells from the tumor at surgery.

Tumor DNA circulating in the plasma might play a role in metastasis. The hypothesis of the genometastasis.

BACKGROUND: Clinical and experimental observations suggest that more than one pathway might be involved in the development of metastases. In the present study, we examined the presence of tumor DNA in plasma using an experimental model in which tumor cells were modified with a genome-associated tag. We also investigated whether plasma of tumor-bearing rats had any effect on cultured cells and healthy animals. METHODS: Transfected cancer cells (DHD/K12-PROb stably transfected with pCDNA3.1CAT.) were injected subcutaneously into the chest of BD-IX rats. Animals were divided into ten groups according to the time between injection of tumor cells and euthanasia. Prior to euthanasia (2-14 week), blood samples were collected by cardiac puncture. To detect circulating tumor cells and CAT-encoding DNA in plasma, we performed PCR with nested primers. Fifty samples of plasma were chosen at random to supplement the medium of fifty cultures of DHD cells for 10-12 days. PCR for the detection of CAT DNA in cells was performed approximately one to two months later. Four healthy rats received an intraperitoneal injection of plasma from a tumor-bearing rat five times at week for 4 to 6 weeks. Animals were sacrificed and samples of liver, kidney, spleen, omentum, blood and lung were processed by PCR for the detection of CAT DNA. RESULTS: Detection of CAT DNA in plasma was slightly more frequent than in the buffy-coat fraction. All surviving cultures that had been supplemented with plasma were positive at some point for CAT DNA. In all four healthy animals injected with plasma of tumor-bearing rats, the marker gene for CAT was found in extracts of lungs. CONCLUSION: Our present observation lead us to propose the following hypothesis. Metastases might develop as a result of transfection of susceptible cells in distant target organs with dominant oncogenes that are present in the circulating plasma and are derived from the primary tumor.

In vitro activation of macrophages by a novel proteoglycan isolated from corms of Crocus sativus L.

Saffron corms contain a proteoglycan that is highly cytotoxic on human tumor cells. The present work was undertaken to study the possible immunomodulatory and anti-invasive properties of this compound. Non-cytotoxic concentrations of this glycoconjugate promoted significant macrophage activation, detected by the release of nitric oxide. A rapid activation of protein kinase C and NF-kappaB was obtained after proteoglycan treatment, which could explain the induction of nitric oxide synthase. Proteoglycan concentrations ranging from 10-1000 ng/ml specifically promoted apoptosis of macrophages, probably triggered by their activation. This molecule did not inhibit in vitro migration or invasion of human tumor cells. Altogether these results support a plausible immuno-modulating activity for this saffron Crocus compound.

Detection of genomically-tagged cancer cells in different tissues at different stages of tumor development: lack of correlation with the formation of metastasis.

Genetic detection of tumor cells in blood, lymphatic nodes or bone marrow using reverse transcription and polymerase chain reaction (PCR) is quite attractive because it allows the early diagnosis of cancer dissemination. Unfortunately, this type of detection strategy cannot be applied to solid parenchymas, because they usually share with tumor cells the mRNA markers. To avoid this impediment, we have developed an experimental model of cancer using cells with a genome-associated tag. DHD/K12-PROb cancer cells were stably transfected with pcDNA3.1CAT. Approximately 10(6) transfected cells (DHD-CAT cells) were injected subcutaneously into the chest of BD-IX rats. Animals were divided into 11 groups according to the time between injection of tumor cells and euthanasia. An additional 'untagged group' was injected with untransfected cells (DHD-Wild). Blood and tissues samples were collected after euthanasia. Macroscopic and microscopic analysis was done. To detect circulating tumor cells or their presence in peripheral organs, we performed PCR with nested primers to amplify chloramphenicol acetyl transferase-encoding (CAT-encoding) DNA sequences. The minimum number of cells that yielded detectable cells routinely was 2 in 10(6). No modification of cancer aggressiveness was observed in DHD-CAT cells. DHD-CAT cells were detected by PCR in lung from the 1st week after inoculation, in liver, spleen and kidney from the 3rd week and in the blood from the 5th week. All animals analyzed 12 weeks after injection showed lung metastases. Metastases in liver, spleen or kidney, either microscopic or macroscopic, were never detected. We have developed an experimental model of cancer based on genomic tagging of tumor cells that allows the detection of small numbers of cells in all organs and the blood. The presence of cancer cells in parenchymas detected with molecular technology does not correlate with the development of clinically relevant metastases.

Effects of long-term treatment of colon adenocarcinoma with crocin, a carotenoid from saffron (Crocus sativus L.): an experimental study in the rat.

We used an experimental model in the rat to examine the effects of long-term treatment with crocin, a glycosylated carotenoid from the stigmas of the saffron crocus, on colon cancer. BD-IX rats were divided into four groups: Groups G1 and G2, designated "cancer groups," were used to study the effects of crocin on the progression of colon cancer, and Groups G3 and G4, designated "toxicity groups," were used to study the effects of the treatment on metabolic processes and the parenchyma. DHD/K12-PROb cells were injected subcutaneously into the chest of Group G1 and G2 animals. From 1 to 13 weeks after inoculation, animals in Groups G2 and G4 received a weekly injection of crocin (400 mg/kg body wt s.c.). Animals in Groups G1 and G3 received no treatment. In addition, lines of animal and human colon adenocarcinoma cells (DHD/K12-PROb and HT-29) were used to perform assays in vitro to examine the cytotoxicity of crocin. Life span was extended and tumor growth was slower in crocin-treated female rats, but no significant antitumor effect was found in male rats. Acute tubular necrosis was found in all kidney samples from crocin-treated animals, but slight signs of nephrotoxicity were found by biochemical analysis of the serum. In assays in vitro, crocin had a potent cytotoxic effect on human and animal adenocarcinoma cells (HT-29 and DHD/K12-PROb cells, 50% lethal dose = 0.4 and 1.0 mM, respectively). Treated cells exhibited a remarkable loss of cytoplasm and wide cytoplasmic vacuole-like areas. In conclusion, long-term treatment with crocin enhances survival selectively in female rats with colon cancer without major toxic effects. The effects of crocin might be related to its strong cytotoxic effect on cultured tumor cells.

Effect of partial HLA class I match on acute rejection in viral pre-infected human liver allograft recipients.

BACKGROUND: Acute rejection in liver transplants is one of the commonest causes of liver dysfunction in the early postoperative period. However, the factors involved in liver graft rejection are still unknown. Our study was aimed at ascertaining whether the degree of HLA class I and class II compatibility or pretransplant viral infection have any influence on early acute liver graft rejection. METHODS: We reviewed clinical and laboratory data in 190 consecutive patients who underwent a liver transplant. HLA-A, HLA-B, and HLA-DR typing for the establishment of an HLA match score was performed by a standard microcytotoxicity method. The existence of pretransplant viral infection was investigated in sera and biopsy tissue by serologic (hepatitis B virus, hepatitis C virus) and polymerase chain reaction (cytomegalovirus) techniques, respectively. The influence of these two factors in acute rejection and the interaction between them was also analyzed. RESULTS: A strong association between viral infection and acute rejection in the group with partial class I matching was found (odds ratio=7.75; P<0.0009), whereas no correlation was observed in the group with zero class I matching (odds ratio=0.98; P=0.81). The rejection percentage in the group in which partial class I match and viral infections coexisted was 60%, whereas in the partially class I-matched group without pretransplant viral presence it was 16%. CONCLUSIONS: These findings suggest a participation of partial HLA class I compatibility in triggering acute rejection in recipients suffering preoperative viral infections and support the idea that HLA class I antigen matching could play a role as a linking element between the MHC-restricted T cell-mediated response to viral infection and the allogenic response in liver transplantation.

HLA-DRB1 and -DQB1 polymorphism in liver recipients: relationship between HLA-DQB1*0302 allele frequency and acute rejection.

Polymorphism of HLA-DRB1 and HLA-DQB1 loci was performed in fifty-three orthotopic liver graft recipients as well as in 108 unrelated healthy controls. Nonradioactive SSOPs were used to study PCR-amplified DNA from peripheral blood lymphocytes and biopsied material. The comparison frequency for DQB1 alleles did not reveal any significant differences between the total group of liver recipients and controls. However, when the liver recipients were subgrouped according to their rejection episode manifestations, increased and significant frequencies were observed for HLA-DQB1*0302 allele in patients showing acute rejection episodes compared to healthy controls or patients without acute rejection. This relationship did not appear influenced by the amino acid beta alanine residue in the 57th position. On the other hand, the study of the DRB1 allele frequencies did not show significant differences in any study. These results suggest that HLA-DQB1 genes could be important in the liver graft alloresponses, opening a way to a better understanding of the special tolerance state, normally observed in this type of transplant, leading us to consider the possible HLA-DQB1*0302 allele effect on tolerance rupture.

CD28 expression on peripheral blood T lymphocytes after orthotopic liver transplant: upregulation in acute rejection.

Despite immunosuppressive treatments, acute rejection remains a significant cause of graft loss. Efficient allorecognition implicates cognate T-cell interactions and requires costimulatory signals such as those delivered via CD28. Therefore, we have studied CD28 peripheral blood T-cell expression, analyzing its possible implications in liver allograft acute rejection. Fifty-five CsA-immunosuppressed orthotopic liver recipients, with or without acute rejection (AR and NAR) were immunocytometrically monitored after transplant and thirty healthy volunteers were studied as controls. In liver recipients the absolute number of CD28+ cells fell sharply immediately after transplant, but no significant differences were detected between the AR and NAR groups either in the absolute number or in the percentage of CD28+ lymphocytes. By contrast, both CD4+CD28+ and CD8+CD28+ T-cell subsets displayed a significant increase in CD28 intensity expression in AR recipients, whereas CD28 expression was significantly downregulated in the NAR recipients. This data suggests that CD28 molecule can be important in the immunologic events preceding acute rejection and that CD28 up- or downregulation could become a useful predictive marker for acute rejection or tolerance development in liver recipients.

[Comparative study of soluble interleukin 2 receptor and adenosine deaminase levels in tuberculous and other etiologies pleural fluids]. / Estudio comparativo de los niveles de receptor soluble de interleucina 2 y adenosín-desaminasa en líquidos pleurales tuberculosos y de otras etiologías.

In order to better understand the immunological mechanisms involved in host protection against Mycobacterium tuberculosis infection, we studied soluble interleukin 2 receptor (sIL-2R) concentration in tuberculous pleural exudates as well as in pleural fluids of non-mycobacterial etiology. We collected pleural fluid from 40 patients: 10 with tuberculous bacterial pneumonia and 10 with trasudate. Soluble IL-2R was measured in the stored specimens using a standard ELISA technique. In patients with tuberculosis, sIL-2R in pleural fluid was 14,666 +/- 5,634 U/ml, significantly higher than was detected in any other group, being 4,341 +/- 2,655 U/ml in pneumonic exudates, 5,542 +/- 3,682 U/ml in neoplastic exudates and 1,377 +/- 125 in trasudates (p < 0.001). Also, an excellent correlation was demonstrated between adenosine-desaminase (ADA) and sIL-2R in tuberculous pleural fluids, with p < 0.001 and r = 0.805. In pleuropulmonary tuberculosis, compartmentalization of the immune response in the pleural space is responsible for the significantly higher levels of sIL-2R that were found in tuberculous pleural liquids compared with the ones detected in other diseases. This observation, as well as the demonstration of a good correlation between sIL-2R and ADA, suggest the possible usefulness of this molecule as an additional marker in the differential diagnosis of pleural effusions, though in the present study it appears to be less reliable than ADA.

A novel functional cell surface dimer (kp43) serves as accessory molecule for the activation of a subset of human gamma delta T cells.

Kp43 is a cell surface molecule expressed by NK cells, gamma delta T lymphocytes, and a small subset of CD56+ alpha beta T cells. In the present study the distribution and function of Kp43 on gamma delta T cells is described. The expression of Kp43 on freshly isolated gamma delta T-cell populations derived from fetal and adult tissues was biased toward memory V gamma 2+/V delta 2+ T lymphocytes and appeared to be up-regulated after activation. Several gamma delta T-cell clones were derived from different sources. Adult V gamma 2+/V delta 2+ T-cell clones expressed the highest levels of Kp43, V delta 1 T-cell clones derived from periphery expressed moderate levels, whereas some thymus V delta 1 T cells and fetal gamma delta T-cell clones displayed low or undetectable levels of Kp43 molecule. Based on the effect of a specific mAb, this dimer appears to be fully functional as previously shown for NK cells. The stimulation of Kp43+ gamma delta T-cell clones with a combination of anti-CD3 and anti-Kp43 co-immobilized on a solid substratum enhanced both the expression of cell surface CD25 and the secretion of TNF-alpha. The restricted expression and functional activity of the Kp43 dimer suggests that it may play an important regulatory role in activation, selection, Ag recognition, and responsiveness of a subset of gamma delta T cells.