A comparison of the use of clomiphene citrate and letrozole in patients undergoing IVF with the objective of producing only one or two embryos.

AIM: The objective of this study was to compare the use of clomiphene citrate and letrozole in an IVF setting in which the objective was to produce only one or two embryos. Either clomiphene citrate or letrozole was used in conjunction with gonadotropins without the use of GnRH antagonists. METHODS: Sixty-two patients received either clomiphene citrate or letrozole with low dose gonadotropins in 128 non-randomized treatment cycles. HCG was given when one follicle was at least 17 mm. Oocyte retrieval was done 34 hours later and fertilization was performed using ICSI. RESULTS: There were no statistically significant differences in the number of large follicles produced, oocytes fertilized, endometrial thickness, clinical pregnancy rates, or delivery rates in patients taking letrozole compared to clomiphene citrate. More mature oocytes were retrieved after clomiphene citrate, but a subset analysis of patients undergoing both treatments did not support this difference. The only statistically different finding in both analyses was the peak estradiol levels during treatment, which averaged 516 pg/ml with letrozole and 797 pg/ml with clomiphene citrate (p = 0.005). Overall, the cancellation rate due to a premature LH surge was 5%. An average of 2.8 mature oocytes were recovered, 2.1 oocytes fertilized and 1.6 embryos were transferred. The overall clinical pregnancy rate per transfer was 25% (95% confidence interval 17.7% to 33.3%) and the overall live birth rate per transfer was 19.2% (95% confidence interval of 11.6% to 26.8%). CONCLUSIONS: Other than peak estradiol levels, there were no clinically significant differences when letrozole or clomiphene citrate was used for mild ovarian stimulation for IVF. There were only rare cancellations because of premature LH surges.

A comparison of the Cook single lumen immature ovum IVM needle to the Steiner-Tan pseudo double lumen flushing needle for oocyte retrieval for IVM.

PURPOSE: This study compared the impact of using the Steiner-Tan pseudo double lumen needle for antral follicle oocyte retrieval to using a conventional non-flushing needle. The Steiner-Tan needle has a much smaller dead space than the needles commonly used for IVM oocyte retrievals. METHODS: This was a retrospective cohort study. The patient population was determined by the time period in which a patient underwent IVM in a single physician's IVF practice. The following data was abstracted from clinical and embryology records: oocytes retrieved, oocytes matured, early maturing oocytes, oocytes fertilized, embryo quality measures, retrieval time, needle punctures, clot formation, and clinical pregnancy rate. RESULTS: The Steiner-Tan needle did not increase the number of oocytes retrieved. It also did not increase the time required for retrieval. However, flushing of antral follicles significantly decreased clot formation in fluid aspirates. Use of the Steiner-Tan needle also significantly decreased the number of vaginal needle punctures during each case. There was a trend toward improved embryo quality, but statistical power was inadequate to show a difference. CONCLUSIONS: The primary benefit of the Steiner-Tan needle was on the embryological aspects of IVM. Decreased blood and blood clots in the aspirates made an IVM retrieval more like conventional IVF for the embryologist. The patient also experienced less tissue trauma without increasing anesthesia or surgical time. There was no improvement in the number of oocytes retrieved, but based on the results, we hypothesized that oocytes were more commonly retrieved from slightly large follicles than when using a routine needle.

Polar body fragmentation in IVM oocytes is associated with impaired fertilization and embryo development.

PURPOSE: The significance of finding a fragmented first polar body in an oocyte prepared for ICSI is controversial with most recent publications suggesting that it is not prognostic for oocyte fertilization or embryo development. Our purpose was to look at this question in the context of oocytes not stimulated for conventional IVF. METHODS: Oocytes obtained for IVM and obtained from follicles at most 12 mm in diameter were evaluated for their polar body morphology soon after they entered metaphase II when they were denuded in preparation for ICSI. Records were evaluated retrospectively for the fertilization rate and the embryo growth rate (cell number) on each day of development for embryos with normal appearing polar bodies or fragmented polar bodies, but no other cytoplasmic dysmorphisms. RESULTS: Oocytes with fragmented polar bodies were significantly less likely to fertilize than oocytes with normal appearing polar bodies (p < 0.0001). Embryos which developed from oocytes with fragmented polar bodies had significantly impaired growth compared to embryos that developed from oocytes with normal appearing polar bodies (p = 0.0328). CONCLUSIONS: Fragmented polar bodies likely reflect cytoplasmic incompetence.

Atrial structural remodeling in patients with atrial chronic fibrillations and in animal models.

Arrhythmia's atrium fibrillation (AF) is the most often met in clinical setting and it is associated with an increased in mortality risk. For profound the structural changes in chronic AF, we are studied the morphological changes of atrium biopsies to be effected at 175 patients. With sustained AF malformative and valvular acquired cardiac diseases operated under extracorporeal circulation. Similar studies we are effected to 11 dogs with partial coronary obstructions to a made periodical EKG investigations. The morphological changes mainly concern accommodation (dedifferentiation) of cardiomyocytes (particularly at experimental model) and mal-accommodation (degeneration of cells with fibrosis replacement features) particularly in acquired valvular diseases. These changes were often interfered. Over study, maintain the hypothesis that the structural changes to be an accommodation more than degenerative response to AF.

Myocardial stunning. Morphological studies in acute experimental ischemia and intraoperatory myocardial biopsies.

Myocardial stunning represent a consequence of brief ischemia with reversible regional contractile dysfunction dependent persist from minute to days after reperfusion, despite the absence of irreversible damage and restoration of coronary blood flow. The evolution of these new ischemic entity were described by experimental acute ischemia and repeated intraoperatory myocardial biopsies effectuated near 200 patients with heart disease, excluding those with cardiac failure and atrial fibrillation. Using histological histoenzymological and particularly ultrastructural methods, only reversible mitochondrial and sarcoplasmatic reticulum lesions, slight glycogen granules depletions and sporadical dissociation of myofilaments by edema were seen. Major mechanism for the state was suggested: generation of oxygen derived free radicals with consequent oxidative stress and impaired calcium homeostasis. Rare morphological appearance on stunned myocardium was signaled in the references, our collective first made these studies in Romania.

Hibernating myocardium, morphological studies on intraoperatory myocardial biopsies and on chronic ischemia experimental model.

UNLABELLED: Hibernating myocardium represent a prolonged but potentially reversible myocardial contractile dysfunction, an incomplete adaptation caused by chronic myocardial ischemia and persisting at least until blood flow restored. The purpose of this study was to investigate the morphological changes and weather relations exist among function, metabolism and structure in left ventricular hibernating myocardium. Material and methods. Experimental study is making on 12 dogs incomplete coronary obstruction during six weeks for morphologic studies of ischemic zones. On 48 patients with coronary stenosis myocardial biopsies was effectuated during aorto-coronarian bypass graft. On 60 patients with valvular disease associated with segmental coronary atherosclerotic obstructions during surgical interventions on a effectuated repeatedly biopsies from ischemic zones. Dyskinetic ischemic areas was identified by angiography, scintigraphy, low dose dobutamine echography to identify the cells viability. On myocardial biopsies various histological, histoenzymological, immunohistochemical and ultrastructural methods were performed. RESULTS: The morphological cardiomyocytic changes can summarized: loss of myofilaments, accumulation of glycogen, small mitochondria with reversible lesions, decrease of smooth reticulum, absence of T tubules, depression of titin in puncted pattern, loss of cardiotonin, disorganization of cytoskeleton, dispersed nuclear heterochromatin, embryofetal dedifferentiation, and persistence of viability. Extracellular matrix is enlarged with early matrix protein such fibronectin, tenascin, fibroblasts. In experimental material the morphological changes present similarities with the human biopsies, but intermixed with postinfarction scar tissue. Redifferentiation of hibernanting cells end remodeling of extracellular matrix is possible after quigle revascularization through aorto-coronary bypass grafts.

Studies on pathogenicity of Yersinia enterocolitica in mice with diabetes mellitus.

In this study, we investigated the colonizing ability as well as the association of Yersinia enterocolitica serotype 0:9 to epithelial cells of the intestinal tract, Peyer's patches, mesenteric lymph nodes, liver, spleen and lungs in Alloxan-induced diabetes mellitus in mice and controls. The results showed that: (a) in diabetic mice the Y. enterocolitica colonizing values were in range of 10(6.5)-10(8.25) CFU/g of feces; (b) maximum colonizing values were found in distal ileum and Peyer's patches and lower in colon; (c) the infection was progressive with dissemination of bacteria in the liver, spleen and lung; (d) in control (non-diabetic) mice, the colonizing values were 10-100 times lower than those found in the diabetic batch; (e) the main histopathological changes noticed, namely ileitis, mesenteric lymphadenitis and septicemia, were presumably induced by high bacterial load in the liver, spleen and lung leading to a septic course of infection as well as toxic effects of heat-stable enterotoxins of Y. enterocolitica (Yst). The results were confirmed by electron microscopy observations. Summing up, these results demonstrate that diabetic mice were more susceptible to Y. enterocolitica cells than normal mice.

Electronmicroscopic and histoenzymological researches regarding myocardial biology in some malformations.

A number of 65 heart malformations, taken from intraoperatory biopsies at different degrees of congestive heart failure were histological, histoenzymological and electronmicroscopical analyzed. It was followed the pathogenesis of the lesions determined by the hemodynamic and hypoxic intracardiac disorders. These hadn't specificity and were disseminated in different evaluative degrees, being predominant in certain myocardial areas. Initially, cardiomyocytes hypertrophical changes appear, and also changes of the edematous matrix. Then, the nuclear lesions, the apoptosis processes and the hypoxic alterations increased to cytolysis. In succession, after the matrix edema was organized, immune lesions and myocardial sclerosis processes appeared. Out of the various lesions, we mention: the sarcoplasmatic fuchsinophilia and alcianophilia, the glycogenic depletion, the decrease of SDH, LDH reactions, the increase of the cytolitic enzymes, especially of the kathepsine B. Out of the ultrastructural changes, we mention: the mitochondrial, sarcotubular lesions and, the lysosomial activations, the interstitial organelles, including the natriuretic granules, synthesized in the ventricular myocytes, details regarding the matrix changes with tendency to fibrosclerosis.

Electronmicroscopic observations regarding the presence of natriuretic granules in the ventricle of patients with cardiopathies.

As part of studies regarding the human myocardium biology in hereditary and acquired cardiopathies, we showed the presence of natriuretic granules (NG) not only in atriums but also in ventricles, in ten of our cases with intraoperative myocardium biopsies. Ultrastructurally, the natriuretic granules occur in lesions produced by hemodynamic and consecutively hypoxic disturbances, being present at the ventricular level, too, both near the Golgi apparatus that secrets them, and diffusely, in cardiomyocytes, beneath the altered organelles and phagolysosomes. Their aspect in similar in cardiac malformations (DSIA. Fallot tetralogy) and in mitral valvulopathies, their abundance being in connection with congestive heart failure (CHF). Although predominant in CHF class I and II, they occur also in the severe decompensations class III and IV, being perhaps felt at the cardiac level as they are released in the blood. This phenomenon is expressed by electronmicroscopic presence of natriuretic granules in the subsarcolema and the increased plasma level of natriuretic peptide, according to biochemical findings reported in the literature. They produce a vasodilating, diuretic, natriuretic effect, contributing to blood pressure regulation and testifying in the neuroendocrine role of human myocardium.

Immunocytomorphological study on the pathogenesis of ankylosing spondylarthritis.

The present investigation is based on the cytomorphological, histopathological (HE, VG, PAS-Alcian, Safranin 0, Gömöri), histoenzymological (acid phosphatase, chondroitinsulphatase, peroxidase) and immunological (rheumatoid factor (RF), circulating immune complexes (CIC), anticolagen II antibodies and C reactive protein (CRP) study on ankylosing spondylarthritis (2.5 cases). The synovial fluid (SF) synoviocytogram showed cytosis (6.067/mm3), with polynucleosis (65.19%) and ragocytosis (17.73%) as compared with the hydrarthrosie SF characterized by lymphocytosis (47%). Enzymological findings revealed phosphatasic and myeloperoxidasic activity in the ragocytary polymorphonuclear (PMNs) and mononuclear cells. Histopathologically, the severe forms of AS correlated with villous chronic synovitis, associated to processes of obliterating vascularitis, fibrosclerosis, necrosis and calcification of disintegrated synovial structures. The articular cartilage was severly damaged, while osseous necrobiosis was noted at the osteocartilaginous junction. Histoenzymologically, the chondrocytes and synovial macrophages showed lysosomal and oxidative enzymatic activity. Immunological assessments (72 sera and 25 synovial fluid samples) showed pathological values of circulating immune complexes, anticollagen antibodies and C reactive protein. Correlation of immunocytomorphological findings demonstrates the involvement of immunologic and enzymatic factors in the pathogenesis of AS.

Morphophysiopathologic studies on the biology of myocardium in acquired valvulopathies.

Myocardial lesions due to various valvulopathies were sampled during open heart surgery under extracorporeal circulation, and histologically, histoenzymologically and ultrastructurally investigated. Various cardiomyocitary and interstitial nonspecific progressive lesions, due especially to hemodynamic and hypoxic alterations, were identified. During the pathogenic stages, we remarked the onset and chronic evolution of perivascular and interstitial oedema, enhanced by a reduced lymph draining. The progressive processes of hyalinization, sclerosis and interstitial fibrosis were subsequent to the interstitial matrix modification. These processes accentuate the myocardial hypoxic lesion due to the nutritional and gaseous exchange alterations. The infrastructure, mitochondrial enzyme equipment, sarcoplasma and tubular network lesions, as well as the intramyocitary oedema that destroys the sarcomeric structure, ended with the activation of lysosome and lysosomal enzymes, giving rise to consequent cytolytic foci. Therefore, the process was extending. In this context, we remarked at the electronmicroscopic examination ventricular natriuremic granules, that are rarely mentioned in literature.

Retroperitoneal extraadrenal paraganglioma report of two cases.

Two cases with retroperitoneal tumours affecting young women who cannot undergo surgery were investigated by biopsy and needle sampling. One of these cases presented a mainly alveolar histologic pattern and was easily diagnosed by immunohistochemical means, as well as a chromaffin paraganglioma. We discussed different diagnoses and approaches in the literature.

Anatomoclinical and bacteriological study on chronic marginal periodontal disease. Therapy with staphylococcic vaccine.

Histological, histochemical and electronmicroscopic investigations were carried out on gum biopsies taken from cases with chronic marginal periodontal disease before and after the treatment with staphylococcic vaccine, thus, morphopathologically proving the findings of Gafar et al. (1985) and of Georgescu et al. (1973; 1974; 1975; 1979; 1995). Before treatment, gum lesions were severe, consisting in an abundance of polymorphic inflammatory infiltrates in the chorion along with oedema and disorganised connective fibres, atrophies and ulceration of the mucosa, purulent deposits at the surface with bacteria and fungi. After the treatment with the staphylococcic vaccine, an epithelial regeneration was seen together with the absence of the chronic inflammation. The intensely fibroparius granulated tissue was also noted, in all cases observing abundant connective fibres, accounting for good results after treatment, notably the firmness of the tooth fitting.

Morphopathological and clinical diagnosis aspects of a bilateral dysembryoplasic renal tumor associated with Bourneville's tuberous sclerosis.

A 34-year old woman was admitted for bilateral flank pain, maleolary and palpebral oedema with an insidious evolution lasting for 5 years. Urogram showed a left ureteral duplicity and a right lacunary pelvic image. MRI and echography indicated the complete modification as a renal structure as well as the presence of some bilateral tumoral formations with a nonhomogenous structure, blood diffusion and adenopathy of the renal hilum. Nephrectomy revealed white-gray tumoral nodular tumours. The histopathological test evidenced proliferations of leiomyofibromatous, angiomatous, lipomatous, cartilaginous types with a benign aspect suggesting the diagnosis of a renal dysembryoplasic tumor (hamartoma).

Immunocytomorphopathological studies on the pathogenesis of rheumatoid arthritis.

Thirty cases of rheumatoid arthritis were submitted to cytomorphological, histopathological (HE, VG, PAS Alcian, Gömöri, Safranine O), histoenzymological (Acid Phosphatase, chondroitin-sulphatase, Peroxidase) and immunological (rheumatoid factor (RF)) studies; circulating immune complexes, anti-collagen antibodies II, Reactive C protein (CRP), Complementary C3 fraction were also assessed. The synoviocytogram of the rheumatoid synovial fluid (SF) indicated a cytosis with polynucleosis and ragocytosis compared to the hydroarthrosic SF defined by lymphocytosis (47.8%). Enzymologically, especially for high titres of rheumatoid factor, a phosphatase and peroxidase activity was observed in polymorphonuclear cells of a ragocytary type and in phagocytic mononuclear cells. The severe forms of rheumatoid arthritis (RA) were correlated histopathologically with chronic villous synovitis associated with some processes of obliterant vascularitis, fibrosis and sclerosis. At the level of synovio-cartilage junction, fissures and a homogenization of the cartilaginous fundamental substance in the vicinity of disintegrated synovial structures were noticed. Histoenzymologically, a lysosomal and oxidative activity was found in chondrocytes and in synovial macrophages. Immunological assessments (73 serum and 60 synovial fluid samples) showed pathological values of circulating immune complexes, anti-collagen antibodies and C reactive protein. The complementary synovial depletion of the C3 fraction underlines the immune character of the rheumatoid synovitis. The immunocytomorphologic data correlation demonstrates the involvement of immunologic and enzymatic factors in the evolution of Rheumatoid Arthritis.

Gastrointestinal stromal tumors--a recently defined entity. Literature data and personal case report.

Stromal tumors of the gastrointestinal tract are controverted tumoral entities which were recently delimited. They were initially identified by immunohistochemical investigation and processing of the gastrointestinal muscular tumors and of the malignant and benign nervous tumors. Those investigations were subsequently amplified by electronmicroscopic studies and observations during the patient's prognostic follow-up. Within these circumstances, monoclonal tumors responsive to smooth muscular antigens (actin, SMA), to polyclonal antigens reacting to S-100, PGP 9.5, NSE and GEAP were identified. Thus, tumors with a nervous origin known as a gastrointestinal nerve tumors or plexosarcomas were differentiated, taking into account the difficulty of their distinguishing from gastrointestinal tumors (GIST) which also shows multinucleated great cells. There were also difficulties in defining bidirectionally differentiated tumors or those nonresponsive to any antigen, all these elements proving their origin from nondifferentiated mesenchymal cells. These data are presented in a personal case report of a 28-year old woman with an acute anteroenteral invagination due to a gastrointestinal stromal tumor, favorably removed by surgery and with a postoperative outcome.

Electronmicroscopic and histoenzymologic study regarding morphophysiopathologic visceral modifications during open heart operations under extracorporeal circulation.

Morphophysiopathologic modifications occurred in myocardium, liver and lung during open heart operations under extracorporeal circulation had been studied using histologic, histoenzymologic, ultrastructural and biochemical investigations. Preexisting myocardic and hepatic lesions subsequent to chronic stasis and hypoxia are accentuated due to the "controlled shock" state induced by extracorporeal circulation. Mitochondrial marker enzymes like SDH, LDH and ATP-ase in the membrane are decreasing their levels and hydrolytic enzymes like acid phosphatase, nonspecific esterase (ANAE) and peroxidase are increasing their activity. These disseminated lesions can become irreversible under the conditions of a severe cardial insufficiency and of a prolonged operation. In the lung, the increase in the hydrolytic enzyme activity suggests the processes of phagocytosis and metabolization of red cells by macrophages.

Histopathological and ultrastructural modifications of the arthrosis articular cartilage.

Thirty samples of articular cartilage taken during the operation from patients with incipient arthrosis, arthrosis with radiological modifications and arthrosis under study for Rheumatoid Arthritis (RA) were investigated using histopathological (HE, VG, PAS-Alcian, Gömöri, Safranine O) and electronmicroscopic techniques. The control material was made of posttraumatic cartilage (Moore prosthesis). Histopathologically, the incipient arthrosis cartilage had superficial exfoliations associated with reduced saframinophilic tinctorial perichondrocytic activity. The arthrosic cartilage with typical radiological modifications was individualized at the synovia-cartilage junction by villous aspects of the synovia associated with perichondrocytic gaps, reduction of safraninophilia and modifications of reticuline-collagenic network. The arthrosic cartilage under study for RA revealed destructive fibrous modifications of the synovia and severe affection of the articular cartilage at synovia-cartilage junction. Electronmicroscopically, the ultrastructural affection was minimum in the incipient arthrosis cartilage developing to chondrocytic degeneration in arthrosis with radiological correspondent. Both histopathological and ultrastructural data emphasize the fact that arthrosis is associated with synovitis following a primitive degenerative process similar to rheumathoid synovitis in arthrosis under study for RA.

Histoenzymological and ultrastructural studies of the macrophage-type pulmonary cell modifications under different chronic stasis conditions.

The optical histoenzymological and electronomicroscopical investigations were carried out en 90 pulmonary biopsies taken from patients under different chronic stasis conditions. To investigate macrophage-type cells we studied the reactions of acid phosphatase and non-specific esterase like enzyme markers, indicating the metabolic state activation degree of this cellular compartment. The ultrastructural examinations revealed the existence of two types of activated pulmonary macrophages. The former is the alveolar macrophage with a prevailing phagolysosome apparatus that pleads for phagocytic function and the latter is the interstitial macrophage with subcellular aspect corresponding to the synthesis and secretion function. The presence of these activated alveolar and interstitial macrophages, as well as their extremely increased number, plead for their direct participation in the appearance and aggravation of the morphophysiopathological disorders in the chronic stasis lungs.