Flow-Cytometric Characterization of Hematopoietic Cells in Non-Pregnant Human Endometrium

Lachapelle M-H, Miron P, Hemmings R, Baron C, Roy DC. Flow-cytometric characterization of hematopoietic cells in non-pregnant human endometrium. Am J Reprod Immunol 1996; 35:5–13 cP Munksgaard, Copenhagen PROBLEM: Immunologic evaluation and quantitation of hematopoietic cells in human endometrium has been difficult to perform, particularly in nonpregnant subjects. In this study, we describe a method for the flow-cytometric characterization of hematopoietic cells present in the endometrium of non-pregnant women. METHOD: Endometrial biopsy samples from normal donors were first mechanically disrupted and filtered to generate a single-cell suspension of leukocyte-enriched endometrial cells. Cells were labeled with a panel of monoclonal antibodies, stained with propidium iodide (PI), and one- or two-color flow-cytometric analysis performed on cells excluding PI. RESULTS: The methodology described in this study was highly reproducible in experiments evaluating the interrun and intrarun variability. We then determined the immunophenotypic profile of endometrial leukocytes from 12 normal females. The majority of leukocytes were T cells (CD3: 47%; CD4: 24%; CD8: 28%) with an important contingent of NK cells (CD56: 32%), the majority of which harbored the unusual CD16-CD56 bright phenotype, and a minority of B cells (CD20: 6%) and monocytes (CD14: 7%). CONCLUSIONS: Flow cytometry can be used to assess antigen expression on the surface of endometrial leukocytes from nonpregnant women. In future studies, it will be possible to use this approach to investigate the role of immune cell populations in the endometrium of patient experiencing reproductive failure.     

The effectiveness of a treatment protocol for male lower urinary tract symptoms in general practice: a practical randomised controlled trial

BACKGROUND: Randomised controlled trials have shown the efficacy of several treatment modalities for lower urinary tract symptoms (LUTS) in selected populations. The effectiveness in daily practice has hardly been investigated, especially in primary care and is dependent on choices between all possible treatment options and best investigated in a comprehensive study, including all treatment modalities (watchful waiting, alpha-blockers, 5-alpha-reductase inhibitors, and surgery). AIM: Assessment of the effectiveness of a comprehensive treatment protocol for LUTS in primary care. DESIGN OF STUDY: Randomised controlled trial. SETTING: Fourteen general practices in the Netherlands. METHOD: Intervention: treatment protocol based on a formalised expert opinion. Control condition: usual care. Study population: 208 subjects with moderate to severe LUTS (IPSS > or =8, median = 13). OUTCOME MEASURES: symptom severity (IPSS [International Prostate Symptom Score]), bother score (Dan-PSS [Danish Prostate Symptom Score]), and maximum urinary flow (Q(max)); incidence of acute urinary retention and urinary tract infections. RESULTS: In the intervention group markedly more subjects used an alpha-blocker at end of follow-up than in the usual care group (24% versus 6%). No significant differences were found between intervention and control group in IPSS, Q(max) or Dan-PSS. CONCLUSION: alpha-blockers and watchful waiting are the most frequent treatment modalities for LUTS in primary care. Our study showed no evidence that a protocol using well-defined indications for all possible treatment modalities based on a formalised expert opinion procedure has added value. Based on our results, we cannot recommend a broadening of the indication for alpha-blockers, which, however, seems to be the current trend.     

Artificial insemination and in-vitro fertilization using donor spermatozoa: a report on 15 years of experience

Donor insemination (DI) using cryopreserved semen commenced at The Royal Women's Hospital in 1976. Over the next 15 years we performed 5953 treatment cycles to achieve 816 pregnancies (13.7% per cycle) and 706 live births. In-vitro fertilization (IVF) using donor spermatozoa commenced in 1986. Over the next 5 years we performed 303 treatment cycles for 185 couples. Including subsequent transfer of cryopreserved embryos, a total of 33% of couples achieved a successful pregnancy by IVF. Statistical analysis indicated that, for DI pregnancies, the most important semen variable was the percentage post-thaw motility, whilst for normal fertilization in IVF it was the pre-freeze motility. These results may be explained by the compensatory effects of post-thaw processing of spermatozoa for IVF, but not for DI in our clinic.      

Histomorphometry and autoradiography of cultured fetal rat long bones

The behavior of fetal rat long bones cultured in vitro according to Raisz's technique (1969) was studied by histomorphometry and autoradiography for a period of four days. The changes were recorded daily both on the trabecular and cortical bone by measuring the bone volume, the number of osteoclasts, and the number of nuclei per osteoclast. Radioactive calcium release was measured and compared to the changes in bone volume and in the number of osteoclasts. An autoradiographic study, using ³H-proline and ³H-thymidine in flash labeling in the medium and ³H-thymidine in follow-up labeling after one injection in vivo was performed to evaluate the bone formation, the cellular proliferation rate and cell differentiation. After four days in culture, an increase in total calcified bone volume was observed which correlatd with changes in the trabecular bone. No significant changes were recorded in the cortical bone. The results showed a good maintenance of the resorption and formation phenomena through an active process of cellular multiplication and differentiation. Undifferentiated cells were labeled in flash label and osteoblast, osteocyte and some osteoclast nuclei were labeled in follow-up studies.     

Severe stenoses and persistent occlusions of the middle cerebral artery: hemodynamic and metabolic consequences studied by positron tomography

Using positron emission tomography and the 15O continuous inhalation technique, we have measured the regional cerebral blood flow (rCBF) oxygen extraction fraction (rOEF) and oxygen consumption (rCMRO2) of non-infarcted tissue in six patients with either tight stenosis (N = 3) or occlusion (N = 3) of the trunk of the middle cerebral artery (MCA); these arterial lesions were shown to be persistent on late and/or repeated angiograms. The patients were studied 1 to 6 months after their last cerebral ischemic event. The data were analyzed in 4 cm2 regions of interest (fig. 1) and were compared to age-matched control values. Regional right/left ratios were tested for significance individually by comparison to 95 p. 100 confidence limits found in control subjects. We found a significant reduction in mean rCBF in the affected MCA territory; concomitantly, there was a lesser decrease in rCMRO2 significant only in the peri-sylvian area; this was associated with a moderate but significant increase in rOEF in the same areas (Table II, fig. 2 and 3). Individually, the reduction in rCBF and the increased rOEF were significant in 5/6 and 2/6 patients, respectively (Table III). These data indicate that rCBF is decreased distal to persistent hemodynamic MCA obstruction in most patients. This hypoperfusion appears due in part to a mild degree of cerebral ischemia (as demonstrated by the occurrence of "misery perfusion"), indicating inadequate perfusion pressure distal to the MCA obstruction. This was associated with a metabolic depression of the cortex possibly resulting from either neuronal loss, or deactivation (diaschisis), or long-standing hemodynamic local failure or any combination of the three.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effects of extra-intracranial arterial bypass on cerebral blood flow and oxygen metabolism in humans

Twelve patients, eleven with a carotid obstruction and one with an occlusion of the middle cerebral artery, were studied before and after a successful unilateral extra-intracranial arterial by-pass, (EIAB) using PET and the 15-0 steady-state technique to measure regional cerebral blood flow (CBF), oxygen extraction fraction and oxygen metabolic rate (CMRO2). In the whole group of patients, both CBF and CMRO2 increased significantly on both cerebral hemispheres after EIAB, returning toward control levels defined in age-matched subjects. Mean oxygen extraction fraction, on the other hand, was not affected. Individually, three different effects of EIAB emerged: 1) Alleviation of a state of long standing unilateral "misery-perfusion", as reported earlier; 2) parallel increase of CBF and CMRO2 bilaterally, which appeared due to improvement of a hemodynamic depression of metabolism, the precise mechanism of which remains obscure; 3) Complex, unexpected changes in the CBF-CMRO2 couple again resulting in increases in CMRO2. This metabolic improvement afforded by EIAB in our patients has not been reported before; it suggests that long-standing hemodynamic failure may induce a metabolic depression that is still potentially reversible by surgical revascularization.     

Processes underlying formal thought disorder in psychiatric inpatients

Three theories about the cognitive processes underlying symptoms of formal thought disorder in psychiatric inpatients were tested. Chapman and Chapman's "excessive yielding to normal bias" theory and a response competition theory were tested by using two ambiguity tasks. Chapman and Chapman's bias theory predicts a smaller ambiguity effect for thought-disordered patients; a response competition hypothesis predicts a larger ambiguity effect. Results showed no difference between thought-disordered and non-thought-disordered patients. To test a distractibility theory of thought disorder, subjects performed a Stroop-type task in which they counted the number of digits (e.g., 3333) or symbols (####) in a set. Thought-disordered patients did show an increased effect of the presence of the digits, and this finding was replicated in a second inpatient sample. Thus, results were consistent with the distractibility theory and with a reformulation of the bias theory, in which the bias shown by thought-disordered patients is not a tendency toward a particular response, but rather toward a particular rule, or set, for responding.     

Focal chronic ischemia and concomitant migraine: an atypical form of Sturge-Weber angiomatosis?

A young man with a left hemifacial hemangioma had during a six months period about forty left hemispheric neurologic attacks suggestive of classic migraine. The neurologic examination was normal during the attack-free period. The CT scan (fig. 1) and the M.R.I. study (fig. 2) only showed a moderate interhemispheric asymmetry. The left internal carotid angiogram showed subtle anomalies of the venous system (fig. 3). All the neurologic manifestations ceased as soon as therapy by aspirin was initiated. A Positron Emission Tomography (PET) study with the oxygen 15 continuous inhalation technique was performed 7 months after the last attack in order to measure the regional Cerebral Blood Flow (rCBF), Oxygen Extraction Fraction (rOEF) and Oxygen Consumption (rCMRO2). Striking, statistically significant, alterations were observed in the left temporo-parieto-occipital area (fig. 4) consisting of a "misery perfusion" syndrome (rCBF = 28-38 ml/100 g/mn; rOEF = 0.64-0.80), without alteration in the rCMRO2 (Table). A repeated PET study 12 months later was unchanged. The association of local chronic oligemia and ipsilateral facial hemangioma, ipsilateral cerebral hypotrophy and venous anomalies suggested the diagnosis of atypical leptomeningeal angiomatosis of the Sturge-Weber type. The importance and persistence of the hemodynamic alterations suggest that chronic oligemia and, hence, tissue hypoxia may participate in the pathogenesis of the migraine-like attacks. Moreover, local circulatory stasis with thrombotic events may be implicated, as suggested by the apparent efficacy of aspirin.