Fresh vs. frozen embryo transfers in patients with KIR Bx haplotype: impact on reproductive outcomes.

OBJECTIVE: A controversy persists over whether or not the type of embryo transfer (ET) influences reproductive outcomes. This study aimed to evaluate the reproductive outcomes of pregnant patients undergoing their first in vitro fertilization procedure and explore the influence of various KIR genotypes on these reproductive outcomes. PATIENTS AND METHODS: Prospective enrollment of patients with infertility who sought treatment at Origyn Fertility Center in Iasi, Romania, was conducted between January 2019 and March 2023. Descriptive statistics and average treatment effects (ATE) using propensity-score matching were employed to analyze our data. RESULTS: Our results indicated that both groups were homogenous regarding baseline characteristics. When we evaluated the ATE of fresh vs. frozen ET on the main outcomes, we discovered that only frozen ET significantly improved the pregnancy rates (ATE: 0.17, 95% CI: 0.04-0.30, p=0.011) and live birth rates (ATE: 0.36, 95% CI: 0.02-1.19, p=0.03). The miscarriage rates were similar between the two groups. None of the evaluated KIR genotypes had a significant influence on the ATE corresponding to fresh and frozen ET. CONCLUSIONS: KIR screening is not necessary before an IVF cycle, except for specific situations such as recurrent pregnancy loss or recurrent implantation failure.

Body composition and circulating estradiol are the main bone density predictors in healthy young and middle-aged men.

PURPOSE: Current fracture risk assessment options in men call for improved evaluation strategies. Recent research directed towards non-classic bone mass determinants have often yielded scarce and conflicting results. We aimed at investigating the impact of novel potential bone mass regulators together with classic determinants of bone status in healthy young and middle-aged men. METHODS: Anthropometric measurements, all-site bone mineral density (BMD) and body composition parameters assessed by dual-energy X-ray absorptiometry and also serum concentrations of (1) the adipokines leptin and resistin, (2) vitamin D and parathormone (PTH), (3) sex hormone binding globulin (SHBG), total testosterone and estradiol (free testosterone was also calculated) and (4) C-terminal telopeptide of type I collagen (CTx) were obtained from 30 apparently healthy male volunteers aged 20-65 years enrolled in this cross-sectional study. RESULTS: Only lean mass (LM) and total estradiol independently predicted BMD in men in multiple regression analysis, together explaining 49% (p ≤ 0.001) of whole-body BMD variance. Hierarchical regression analysis with whole-body BMD as outcome variable demonstrated that the body mass index (BMI) beta coefficient became nonsignificant when LM was added to the model. Adipokines, fat parameters, testosterone (total and free), SHBG, PTH and vitamin D were not independently associated with BMD or CTx. CONCLUSIONS: The present study shows that LM and sex hormones-namely estradiol-are the main determinants of bone mass in young and middle-aged men. The effects of BMI upon BMD seem to be largely mediated by LM. Lifestyle interventions should focus on preserving LM in men for improved bone outcomes.

A rare case of double recurrent choroidal melanoma, with distinctive immunohistochemical features.

A 57-year-old woman, with left choroidal melanoma treated by laser photocoagulation and a history of repeated vitrectomies, checked for left eye acute pain and foreign body-like sensation, symptoms that occurred after three years since the primary tumor treatment. The left eyeball was enucleated and the tissues were investigated by immunohistochemistry for markers associated with cell differentiation, proliferation and adhesion, cell cycle regulation, apoptosis control, vascularization, invasiveness and local immune response. We identified, in fact, two independent tumors, with different localization and sharing some common features, markers of a highly aggressive potential: loss of cell differentiation markers and cell cycle regulators, ability to avoid death by suppressing Fas antigen expression and important invasive capacity by down regulation of E-cadherin expression. However, only in the posterior tumor, we found cells with high proliferation rate, Fas ligand molecule expression and MMP-9 secretion, acquisitions associated with a much more aggressive behavior. These particular phenotypes allowed the posterior cells to grow and to invade the surrounding tissues more rapidly than the anterior ones, leading to the development of a large size tumoral mass, responsible for the clinical symptoms. Photocoagulation, by destroying the tissues, makes impossible the evaluation of the primary tumor's biological features, important for the tumor evolution. The absence of these data stresses the importance of patient monitoring, eventually addressing a panel of soluble markers associated with recurrence or metastasis development.

[Immunohistochemical evaluation of inflammatory reaction in a case of pilomatrixoma]. / Evaluarea prin imunohistochimie a reactiei inflamatorii intr-un caz de pilomatricom.

OBJECTIVES: To identify the types of the cells in a case of pilomatricoma and to evaluate the lesion's stage, the cells' proliferating rate and the associated inflammatory reaction. MATERIAL AND METHODS: The paraffin-embedded tissue was investigated by histological examination and by immunohistochemistry for the expression of some markers such as: cytokeratins, CD3, CD20, CD68, PCNA, CD34 II. RESULTS: The lesion presented the characteristic epithelial cells of a classical pilomatricoma: bazaloid cells, ghost cells and transitional cells. 10-15% of the bazaloid cells were PCNA+. The MNF 116 antibody labeled only some of the transitional and of the ghost cells. We found no CD3-positive cells and few CD20-positive cells. A marked inflammatory reaction was noticed, dominated by giant multinucleated cells, positive for CD68 and PCNA and a rich network of blood vessels with a high vascular density. CONCLUSION: The histological pilomatricoma diagnosis was straightforward on the basis of the bazaloid and ghost cells presence. Immunohistochemistry brought additional data regarding the cells proliferation rate, the stage of the lesion and the intensity of the associated inflammation.

[Contribution of immunohistochemistry and flow-cytometry in the study of endometrial pathology]. / Aportul imunohistochimiei si flow-citometriei in studiul patologiei endometriale.

Endometrioid endometrial carcinoma is characterized by hyperestrogenism and high sensitivity to progestogens. Our study was designed in order to demonstrate the hormonal influences in neoplastic endometrium, by investigating steroid receptors expression and their correlation with proliferation activity, with matrix enzymes expression, and with susceptibility to apoptosis inducers, from hyperplasia to carcinoma. 13 cases were included in our study, 7 endometrial hyperplasias, and 6 endometrial carcinomas. Immunohistochemistry technique was performed, using antibodies against ER-á, PR, PCNA, MMP-2, MMP-9, Fas, and FasL molecules. Flow-cytometry was complementary used, for steroid receptors, PCNA and MMPs. Endometrial hyperplasias were positive for both hormonal receptors, in epithelial and stromal cells. An evident decrease of the percent of positive cells and of the staining intensity of both ER and PR was observed in poorly differentiated endometrial carcinomas. Endometrial hyperplasias presented a similar proliferative index with differentiated endometrioid endometrial carcinomas. Poorly differentiated endometrial carcinomas showed the highest PCNA index. Both types of investigated MMPs were evident, with similar aspect of localisation for both MMP-2 and MMP-9. The staining intensity for MMP-9 was higher than that of MMP-2, and was identified both in epithelial and in stromal cells. Fas and FasL expressions were identified in glandular epithelium of endometrial hyperplasias and carcinomas, although the staining intensity was reduced. Flow-cytometry showed a correlation between qualitative and quantitative data concerning hormonal receptors, PCNA and MMPs. Our study emphasises that neoplastic endometrial cells express several molecules correlated with malignant transformation and tumoral progression, by coordinated intervention of steroids, proliferating factors, gelatinases, in opposition with systems involved in apoptosis initiation.

[Connective tissue growth factors, CTGF and Cyr61 in drug-induced gingival overgrowth--an animal model]. / Factorii de crestere ai tesutului conjunctiv, CTGF si Cyr61, in hipercresterea gingivala indusa medicamentos. Model experimental animal.

Human gingival overgrowth may occur as a side effect of chronic administration of some therapeutic agents. The mechanisms responsible for the gingival tissues lesions, fibrosis and inflamation, involve an impaired balance between the production and the degradation of type I collagen. It has been demonstrated that CCN2/CTGF, a connective tissue growth factor, is highly expressed in the gingival tissues and positively correlated with the degree of fibrosis in the drug-induced gingival overgrowth. The aim of this study was to identify the presence and localization of CCN2/CTGF and CCN1/Cyr61, members of the same molecular family, in gingival tissues of cyclosporin A- and nifedipine-treated rats, by immunohistochemistry. Staining was evaluated with light microscope and the results show cellular and extracellular CTGF in nifedipin gingival overgrowth tissues with intensity of labeling higher compared to the CsA gingival overgrowth tissues or the controls. The staining for Cyr61 shows its intracellular localization with no diference of labeling intensity between drug-induced gingival overgrowth and normal tissues. Also, we were interested in the gingival TGF-â expression in those animals. We didn't find any commercial anti-rat TGF antibody and our anti-human antibody shows no cross-reactivity with rat tissues. The data from our study sustain the involvement of CTGF and Cyr61 as growth factors in the gingival tissues and the CTGF association with drug-induced gingival overgrowth.

[High values of soluble cytokeratin 18 in breast carcinomas may have different meanings]. / Valorile crescute ale citokeratinei 18 serice in neoplasmele mamare pot avea semnificatii diferite.

We have investigated the cellular and serum CK18 in 26 non-treated primary ductal invasive breast carcinomas. The soluble CK18 (TPS) was detected by chemiluminescent assay, and the cellular CK18 and PCNA expression by immunocytochemistry. Flow-cytometry was used to estimate the amount of DNA in malignant cells. There was a significant correlation between soluble CK18 and the pre-menopausal status (p < 0.05), characterized in our group by a PCNA estimated low proliferation index. We have also found a significant correlation between soluble CK18 and the DNA index (p < 0.01). The intracellular CK18 has correlated with the PCNA expression (p < 0.05), while no correlation could be found between cellular and serum CK18. The values of soluble CK18 may offer information about the treatment-induced cell death, if monitored, while isolated measurements should be interpreted cautiously. Elevated levels of serum CK18 in non-treated carcinomas may rather reflect a high tumor turn-over or perhaps a more intensive tumor cell killing.

[Different phenotypes generated by changes in bcl-2 and p53 expression in breast cancer]. / Fenotipuri diferite generate prin modificari in expresia bcl-2/p53 in cancerul mamar.

We have identified by immunohistochemistry/ immunocytochemistry the expression of bcl-2 molecule in 55 primary breast carcinomas and in 30 corresponding axillary lymph nodes metastases, together with a set of molecules known as prognostic factors: estrogen receptors, progesterone receptors, and p53 protein. Our results demonstrated a significant correlation (p < 0.05) between bcl-2 and hormonal receptors expression in tumors, but not in axillary metastases (p < 0.1), a significant inverse correlation between bcl-2 and p53 expression in primary tumors (p < 0.02), but a significant direct correlation in axillary metastases (p < 0.02). The bcl-2+/p53- phenotype, associated with normal breast epithelium, is present in 79.17% primary tumors, but only in 15.38% axillary lymph nodes metastases. A larger number of lymph nodes metastases expressed a bcl-2+/ p53+ more aggressive phenotype compared with primary tumors (58.82% versus 48.39%). This shows that changes in the expression of bcl-2, p53, estrogen and progesterone receptors can lead to an increased cellular aggressiveness and thus to an increased tumoral invasive and metastasizing potential.

[Evaluation of the angiogenic process in breast carcinoma--correlations with cancer aggressiveness]. / Evaluarea angiogenezei în cancerul mamar. Corelatii cu agresivitatea tumorala.

Angiogenesis represents an essential event required by tumors to support their growth. The aim of our study was to investigate the presence of neovascularization in 44 primary breast carcinomas and in 24 axillary lymph nodes metastases and to establish a possible correlation between the presence of tumor angiogenesis, some clinical and pathological features of the cases and the expression of p53, an important cell cycle regulator. To identify the new blood vessels, we used immunohistochemistry for von Willebrand factor, a marker of the endothelial cells. The results showed that 77.27% of the primary breast carcinomas and 75% of the lymph nodes metastases are positive for von Willebrand factor and this positivity is significantly correlated (p < 0.05) with the expression of p53, supporting the idea that angiogenesis is a marker for tumor aggressiveness and p53 could be involved in this process.

[Immunohistochemical evidence of chronic inflammation in acne vulgaris]. / Evidentierea imunohistochimica a celulelor caracteristice inflamatiei cronice în acneea polimorfica juvenila.

The etiology and pathogenesis of acne vulgaris are not yet completely understood. Therefore we have investigated 5 patients with different clinical forms of disease, including the rare form of acne fulminans. Taking into consideration the four factors that are currently incriminated in the development of acne, sebaceous hypersecretion, hyperkeratosis of the pilosebaceous infundibulum, bacterial colonisation and perifollicular inflammation, we have focused our study on a set of cells involved in the chronic inflammatory process. We have evidenced by immunohistochemistry methods, using appropriate monoclonal antibodies, the presence of T lymphocytes and macrophages, while the B cells could be evidenced only in the severe forms. We were also interested to investigate the occurrence of new capillary formation, as an accompanying phenomenon of the inflammatory process. The presence and histological distribution of these cells highly supports the hypothesis that the mechanisms underlying the development of acne vulgaris belong to the Delayed Type Hypersensitivity.

[The significance of Fas/FasL molecular system in breast cancer]. / Semnificatia expresiei sistemului molecular Fas/FasL în cancerul mamar.

Fas (CD95/APO-1) and its natural ligand, FasL, are molecules expressed on cellular membranes, being involved in the induction of programmed cells death or apoptosis. Recently, it has been shown that malignant cells originating from solid tumors tend to inhibit the expression of Fas, as an escape mechanism from the immune cells' attack and to express FasL, as a counterstrike mechanism against the immune effector cells. The purpose of this study was to investigate, by immunohistochemistry, the presence of Fas and FasL in 15 breast carcinomas and to establish possible associations between the expression of these molecules and the histological type and grading of the tumors. Our results showed that 7 breast tumors have lost the expression of Fas and 11 tumors were positive for the Fas-ligand expression, important arguments for the mechanisms of immune escape and tolerance induction. Furthermore, 7 of the 11 FasL+ tumors were poorly differentiated invasive ductal carcinomas, suggesting a possible association between FasL expression and tumor aggressivity.

Sequential phenotype changes in progressive lesions of breast carcinoma.

Malignant transformation is the result of genetic events, translated into sequential acquisitions of multiple abnormalities in the control of cellular growth and cell cycle regulation. We determined the expression of the estrogen receptor (ER), progesterone receptor (PR), c-erbB-2 and p53 gene products in a patient with mixed (ductal and lobular) invasive breast carcinoma bearing different coexisting lesions. The purpose of the study was to establish a possible correlation between the expression pattern for these molecules and the histological appearance of the breast tumor. Our results showed no positivity for ER. PR expression was restricted to normal epithelium, simple hyperplasia and in situ carcinoma. c-erbB-2 was detected in all lesions with the exception of normal epithelium and immunostaining for p53 was found positive only in in situ and invasive carcinoma. These findings support the hypothesis of tumorigenesis as a multistep process and as a sum of changes, each representing an advantageous acquisition for the malignant cells' behavior. The loss of hormone receptors' expression occurred as an early event in this case, while the p53 mutations were found only in more advanced neoplastic lesions.

[Overexpression of c-erbB-2 gene product is associated with poor prognosis factors in breast carcinoma]. / Supraexpresia produsului genei c-erbB-2 se asociaza cu factori de prognostic rezervat în carcinomul mamar.

Oncogenes, the abnormal forms of proto-oncogenes, were shown to be involved in malignant transformation and in tumor progression. c-erbB2/HER2/neu is member of EGFR family and encodes the p185 protein, which functions as a tyrosine-kinase. Gene amplification and/or p185 overexpression were reported to be associated with poor prognostic in cancer. Our purpose was to investigate p185 immunohistochemical expression in breast carcinomas and in the corresponding axillary lymph nodes metastases and to identify possible correlation between p185 and other factors of poor prognostic, such as loss of hormonal receptors expression. In our study, 40.91% of cases were erbB-2 positive, p185 expression being maintained from the primary tumors to axillary metastases and associated with positive nodal status and with the absence of hormonal receptors expression (p < 0.05). These findings support the hypothesis the c-erbB2 is an advantageous acquisition for the aggressive behavior of the tumor cell and for its ability to invade and metastasize.

[Loss of hormonal receptors expression from primary breast carcinoma to axillary lymph node metastases]. / Pierderea expresiei receptorilor hormonali de la tumora primara la metastaza ganglionara în carcinomul mamar.

Breast tumors growth is regulated by female sex steroid hormones. The level of the estrogen and progesterone receptors (ER and PR) expression by the malignant cells is important for the evaluation of the tumor prognostic and the benefit of a hormonal therapy. The aim of our study was to identify the expression of estrogen and progesterone receptors in primary breast tumors and in the corresponding axillary lymph nodes metastases, in 24 cases. The results showed that more than 30% of poorly differentiated breast carcinomas lost their expression of hormone receptors from the primary tumors to axillary metastases, an event which can be associated with an aggressive tumoral behaviour and resistance to hormonal therapy.

[Intracellular events in T lymphocyte's activation]. / Evenimente intracelulare în activarea limfocitului T.

Adaptive immune responses are not initiated at the site where a pathogen first establishes a focus of infection. They occur in the organized peripheral lymphoid tissues, to which the pathogen or its products are transported, trapped, captured by specialized cells, called antigen-presenting cells (APC), which process and present the antigen to T lymphocytes. Activation of naive T cells requires two signals: the first signal is represented by the specific recognition of a foreign peptide fragment bound to a self MHC molecule, but this is not enough. The second signal, called co-stimulatory signal is represented by other molecules, expressed on the membrane or secreted by the APCs for which T cells express specific ligands. Binding of antigen to the T-cell receptor initiates a series of biochemical changes within the T cell, involving a large number of molecules.

[Detection of lymph nodes micrometastases in breast carcinoma using immunohistochemistry for cytokeratin 8]. / Depistarea micrometastazelor ganglionare în carcinomul mamar prin imunohistochimie pentru citokeratina 8.

The lymph nodes status is one of the most important prognostic factors in breast cancer. The routine hematoxilin-eosin staining is efficient for the metastases detection only when there is a large number of tumor cells, while a small number of metastatic cells can easily remain undetectable. For those situations, the immunohistochemistry for cytokeratins, markers of the epithelial cells, is a very sensitive method. We have investigated the cytokeratin 8 expression in 10 primary breast carcinomas and in the corresponding axillary lymph nodes, comparing with hematoxilin-eosin. The routine examination has detected axillary lymph nodes metastases in six cases, confirmed by the immunohistochemistry for cytokeratin 8. Four cases were diagnosed as negative for the axillary lymph nodes metastases by the hematoxilin-eosin staining. In all those four cases, immunohistochemistry for cytokeratin 8 has detected a small number of tumor cells, either spread in the lymph nodes tissues, either confluent as small islets.

Low p27(Kip1)expression is associated with poor prognostic factors in breast carcinomas.

PURPOSE: This study was designed to detect the presence of the p27(Kip1) protein expression in breast carcinomas and corresponding axillary lymph node metastases, as well as any potential correlation of p27(Kip1) with factors of tumor progression and prognosis (estrogen receptors-ER, progesterone receptors-PR, c-erbB-2, p53). PATIENTS AND METHODS: 44 primary breast cancers and positive axillary lymph nodes from 24 cases were examined by immunohistochemistry. RESULTS: 19 out of 44 (43.18%) tumor specimens expressed the p27 protein, whereas 25 (56.82%) specimens expressed it only at low levels or were negative. Absence of p27 correlated significantly (p < 0.05) with a negative status for ER and PR receptors. The presence of p27 protein in primary tumors was always associated with positive expression in the corresponding nodal metastases. CONCLUSION: Our results indicate a correlation between lack or low levels of p27 protein expression and the absence of hormone receptors. The p27 phenotype is preserved from the primary breast tumors to the corresponding axillary lymph node metastases.

Crystal structure of a T cell receptor Valpha11 (AV11S5) domain: new canonical forms for the first and second complementarity determining regions.

We describe the X-ray crystallographic structure of a murine T cell receptor (TCR) Valpha domain ("Valpha85.33"; AV11S5-AJ17) to 1.85 A resolution. The Valpha85.33 domain is derived from a TCR that recognizes a type II collagen peptide associated with the murine major histocompatibility complex (MHC) class II molecule, I-A(q). Valpha85.33 packs as a Valpha-Valpha homodimer with a highly symmetric monomer-monomer interface. The first and second complementarity determining regions (CDR1 and CDR2) of this Valpha are shorter than the CDRs corresponding to the majority of other Valpha gene families, and three-dimensional structures of CDRs of these lengths have not been described previously. The CDR1 and CDR2 therefore represent new canonical forms that could serve as templates for AV11 family members. CDR3 of the Valpha85.33 domain is highly flexible and this is consistent with plasticity of this region of the TCR. The fourth hypervariable loop (HV4alpha) of AV11 and AV10 family members is one residue longer than that of other HV4alpha regions and shows a high degree of flexibility. The increase in length results in a distinct disposition of the conserved residue Lys68, which has been shown in other studies to play a role in antigen recognition. The X-ray structure of Valpha85.33 extends the database of canonical forms for CDR1 and CDR2, and has implications for antigen recognition by TCRs that contain related Valpha domains.

[Lymphocyte traffic in the events that follow lymphocytes activation in the germinal centre]. / Traficul limfocitar si evenimentele ulterioare activarii limfocitelor în centrul germinal.

After their maturation, lymphocytes migrate from the primary lymphoid organs to the secondary lymphoid organs and tissues. Once in the secondary tissues the lymphocytes do not simply remain there; many move from one lymphoid organ to another via the blood and lymph. This process allows a large number of antigen-specific lymphocytes to come in contact with their appropriate antigen in the microenvironment of the peripheral lymphoid organs and this is important since lymphocytes are monospecific and only a limited number of cells are able to recognize and interact with a particular antigen. When B cells are activated by antigen, with the help from the T cells, they mature either into AFCs (Antibody-Forming Cells), or they develop into memory cells. The germinal centers in various lymphoid tissues represent the sites of the immune response development and memory B cells generation. At these sites, the B cells undergo active hypermutation of the variable genes, a process that can lead to death by apoptosis for some cells.

A role for the region encompassing the c" strand of a TCR V alpha domain in T cell activation events.

The distinct strand topology of TCR V alpha domains results in a flatter surface in the region encompassing the c" strand than the corresponding region in Ig V domains. In the current study a possible role for this region in T cell activation has been investigated by inserting a potential glycosylation site at V alpha residue 82. This residue is in proximity to the c" strand and distal to the putative interaction site for cognate peptide:MHC ligand. An additional N-linked carbohydrate at this position would create a protrusion on the V alpha domain surface, and this may interfere with TCR aggregation and/or recruitment of signaling molecules. The modified TCR has been expressed in transfected T cells, and the phenotype following stimulation has been compared with that of cells expressing the wild-type TCR. The mutation has significant effects on activation-induced cell death and TCR internalization, but, unexpectedly, does not affect IL-2 secretion. Furthermore, analyses with tetrameric, peptide:MHC class II complexes suggest that the mutation decreases the ability of the TCR to aggregate into a configuration compatible with avid binding by these multivalent ligands.