[Three-dimensional imaging in central serous chorioretinopathy]. / Dreidimensionale Bildgebung bei Chorioretinopathia centralis serosa.

BACKGROUND: Conventional optical coherence tomography (OCT) has brought new insights to retinal diagnostics, but it is restricted due to its low scanning speed and limited resolution. In this study, high-resolution raster scanning OCT (HR-OCT) was used to identify typical changes in central serous chorioretinopathy (CSC). METHODS: Fifteen consecutive patients with acute CSC were imaged using HR-OCT with an axial image resolution of 6 mum. The scanned area measured 6x6 mm2. Three-dimensional (3D) evaluation was performed using various analysing programmes. RESULTS: Topographic changes in CSC could be visualised with 3D reconstructions in all locations. Retinal thickness was measured with the automatic segmentation mode and quantified precisely. Results were presented in 2D and 3D maps. En-face imaging as a modality to present an integrative overview showed specific alterations in CSC. CONCLUSION: HR-OCT was able to provide essential additional information about CSC when combined with appropriate analysing programmes, allowing the identification of typical differences. Aside from precise volumetric measurements, exact localisation of pathological deviations could be achieved.

Trimethoprim-Sulfamethoxazole-induced Hepatotoxicity in a Renal Transplant Patient.

Drug-induced liver injury (DILI) represents liver damage from various therapeutic drugs. Antimicrobials are among the most common causes of DILI. We report a case of hepatic toxicity due to Trimethoprim-sulfamethoxazole (TMP-SMX) in a patient who underwent renal transplantation. Diagnosis has been made after a careful history taking, exclusion of competing etiologies and reversal of biochemical abnormalities after withdrawal of the antibiotic. TMP-SMX liver toxicity is well known but remains unpredictable and is rarely reported.

Alpha 2-macroglobulin production by the human endometrium.

alpha 2-Macroglobulin (A2M) is a broad spectrum plasma protease inhibitor previously described in uterine effluents and recently demonstrated to bind and possibly modulate the functions of cytokines. As cytokines, proteases, and protease inhibitors are important in implantation and endometrial physiology, we sought to investigate and characterize the pattern of production of A2M in the endometrium. Endometrial tissues from different phases of the menstrual cycle were analyzed for A2M production. Tissues were incubated in methionine-free Minimum Essential Medium with [35S]methionine for 12 h at 37 C in 5% CO2. Conditioned media were immunoprecipitated with a polyclonal antibody to human A2M. Recovered proteins were resolved under reducing and nonreducing conditions by 5% sodium dodecyl sulfate-polyacrylamide gel electrophoresis and analyzed by autoradiography. Immunohistochemistry was performed on both cryostat sections of OCT-embedded tissue and formalin-fixed paraffin-embedded tissue. The intensity of staining was evaluated, and a histochemical score was assigned. Comparisons between histochemical scores were performed using the nonparametric Wilcoxon rank sum test. Immunoprecipitation with A2M antibody yielded a single 320-kilodalton protein band under nonreducing conditions and a single 182-kilodalton band under reducing conditions. Both physical and immunological properties of the recovered protein were consistent with A2M. A2M was identified in all endometrial samples and represented approximately 2% of the total radiolabeled proteins produced. Immunohistochemical analysis revealed prominent stromal, but no glandular, staining for A2M throughout the menstrual cycle. The stromal staining in secretory endometrial samples was significantly more intense than that in proliferative samples. In the proliferative phase, staining was more intense in the spongiosum and basalis layers, and less intense in the superficial compactum layer. In the secretory phase, it remained very intense in the spongiosum, but less so in the basalis layer. These findings indicate that A2M is predominantly produced by the stromal component of endometrial tissue. This production is menstrual cycle dependent, with zonal differences in A2M expression within the endometrium, which may indicate a functional significance relevant to the process of implantation that merits further investigation.

Endometrial alpha-2 macroglobulin; localization by in situ hybridization and effect on mouse embryo development in vitro.

alpha-2 macroglobulin (A2M) is a 718,000-kDA broad spectrum plasma protease inhibitor whose production by the human endometrium was recently reported. The multifunctional A2M receptor, also known as low-density lipoprotein receptor-related protein, was also recently immunolocalized to the endometrial stroma. The objective of this study was to further characterize the endometrial site of expression of A2M, and to study its effects on mouse embryo development in vitro, to gain some insight into the functional significance of its endometrial production. Formalin-fixed, paraffin-embedded human endometrium from hysterectomy and endometrial biopsy specimen was used for in situ hybridization analysis, with 35S-labeled riboprobes representing subcloned A2M complementary DNA (cDNA) fragments. Duplicate sections of human endometrium were hybridized with sense and antisense probe and coated with photographic emulsion. Resultant autoradiograms were analyzed qualitatively by light- and darkfield microscopy and quantitatively by a computerized analysis of the signal intensity. Immunohistochemistry and immunoblotting for endometrial tissues were performed using an affinity-purified polyclonal antibody to human A2M. The effect of A2M on mouse embryo development was studied by exposure of one cell mouse embryo in culture to physiological concentrations of biologically active and inactive A2M. Expression signals for A2M were more numerous and intense in the secretory endometrium, compared with proliferative endometrium. Endothelial cells lining the endometrial blood vessels seemed to be the main source of A2M expression. The A2M expression signals in secretory endothelium were 2- to 3-fold stronger than the proliferative endothelium, suggesting transcriptional activation of A2M expression in the secretory endothelium. Glandular expression was observed in secretory endometrium from two patients with endometriosis. Ectopic endometrial tissues also produced A2M. A2M at concentrations of 400-500 mumol/L significantly inhibited blastocyst development of mouse embryos in vitro. A2M is expressed predominantly by the endometrial endothelial cells and may be involved in endometrial physiology. Physiological concentrations of A2M inhibit mouse embryo development in vitro, suggesting that endometrial production of A2M may play a role in regulating preimplantation embryo development.

Localization of the expression of complement component 3 in the human endometrium by in situ hybridization.

C3 production by the human endometrium has been previously described. The objective of the current study was to localize the site of expression and regulation of the third component of complement, C3, in the endometrium. Eight secretory and eight proliferative archival endometrial samples from hysterectomy and endometrial biopsy specimens were used for in situ hybridization analysis. This analysis was performed with a radiolabeled riboprobe synthesized from a 736-bp template representing sequence 1944-2680 of the human C3 complementary DNA. Duplicate sections were hybridized with sense and antisense riboprobes. Resultant autoradiograms were analyzed qualitatively by light- and darkfield microscopy. In proliferative endometrium, minimal expression of C3 was observed and was limited to a few stromal patches and glands throughout the section. In the secretory samples, prominent C3 expression was observed in both the glands and stroma of the basalis layer. Endometrial lymphocytes did not express C3. Endometrial stromal and glandular cells express the C3 gene. Endometrial lymphocytes did not express C3, but other nondistinct lymphoid elements scattered in the stroma may be expressing C3. There was a visibly more intense expression of C3 in the basalis layer of the secretory endometrium than in proliferative endometrium. The spatial and temporal pattern of C3 expression may have implications in normal menstrual physiology and in the immunological response of the endometrium to the invading trophoblast during placentation.

Impact of hormone replacement therapy on the body mass and fat compositions of menopausal women: a cross-sectional study.

OBJECTIVE: The purpose of the study was to compare the body mass and fat compositions of menopausal women who were taking conventional doses of hormone replacement therapy (HRT) with that of menopausal women who were not taking any hormones. DESIGN: The body fat composition of 169 healthy postmenopausal women was measured using a noninvasive handheld machine, the Electrolipograph (BioAnalogics ELG, Beaverton, OR, USA). Impedance to electrical flow in tissues is lower with increasing water content of the tissue. Information on HRT, lifestyle, diet, smoking, and alcohol was obtained from the medical record and by a telephone interview before women were invited to participate. HRT and non-HRT groups were compared. Multivariate linear regression, which included age, years since menopause, type of menopause, and use of HRT, was performed for each of the two major outcomes: body mass index (BMI) and percentage of body fat. RESULTS: Comparisons between subgroups showed a large number of significant differences reflecting differences in age since menopause, baseline BMIs, and baseline waist to hip ratios. In the regression model, however, the only factor significantly associated with lower fat and BMI was the use of HRT. Women who were taking HRT had significantly lower percentages of body fat (-4.8%; p < 0.001) and BMI (-2.6 kg/m2; p < 0.001) compared with nonusers. Age and duration and type of menopause were not significant predictors of weight and BMI in this group of postmenopausal women. CONCLUSIONS: In this study, HRT seems to be associated with a significant reduction in postmenopausal weight and fat mass gains. This may be an important mechanism by which HRT exerts its beneficial long-term effects on cardiovascular health.

Disease activity of hepatitis C correlates with single-stage polymerase chain reaction detection of hepatitis C virus.

Hepatitis C virus (HCV) RNA was amplified and detected in the serum of 17 anti-HCV antibody positive patients using a single-stage (30 cycles) polymerase chain reaction (PCR). Specific amplification, targeted to the C-100 protein (anti-HCV) region, was confirmed by direct sequencing of the PCR product. Single-stage PCR detected the virus in 11 patients. Polymerase chain reaction-positive patients demonstrated a significantly higher histologic activity index (10.3 + 1.2 standard error of mean [SEM] than those testing negative (5.8 + 1.5 SEM, P < .05). Nine of 11 PCR-positive patients exhibited a rise in alanine transaminase (ALT) values within 1 month of assay, compared with only 1 of 6 PCR-negative patients. The correlation between rising ALT levels and PCR positivity was significant (P < .01). Direct sequencing revealed mutability in all cases, some of which resulted in amino acid substitutions. The authors concluded that HCV detection using single-stage PCR correlates with biochemical and histologic features of disease activity. Mutability is likely an important feature of HCV pathobiology and may significantly affect detection methods.

The human "peripheral-type" benzodiazepine receptor: regional mapping of the gene and characterization of the receptor expressed from cDNA.

A cDNA for the human "peripheral-type" benzodiazepine receptor (PBR) was isolated from a liver cDNA library. The 851-nucleotide probe hybridized with a approximately 1 kb mRNA in Northern blots of RNA extracted from various human tissues and cell lines. The human PBR probe was hybridized to DNA from a somatic cell hybrid mapping panel to determine that the gene maps to chromosome 22. With a regional mapping panel for chromosome 22, we localized the gene within band 22q13.31. The ligand-binding properties of the receptor expressed from the cDNA were examined in transient expression experiments and compared to the endogenous human PBR. The PBR ligand [3H]PK 11195 had high affinity for the expressed receptor in COS-1 cells, but the affinities of a pair of isoquinoline propanamide enantiomers differed remarkably in expressed and endogenous human PBR. These findings reveal that the host cell and/or post-translational modification may have an important influence on PBR function.

The effect of a carbohydrate-rich beverage on mood, appetite, and cognitive function in women with premenstrual syndrome.

OBJECTIVE: To test the efficacy of a specially-formulated, carbohydrate-rich beverage (one known to increase the serum ratio of tryptophan to other large neutral amino acids) on the mood, cognitive, and appetitive disturbances of premenstrual syndrome (PMS). METHODS: Twenty-four women with confirmed PMS were enrolled in a double-blind, crossover study to test the efficacy of the specially-formulated beverage compared with two other isocaloric products on PMS symptoms. The study was conducted over three menstrual cycles preceded by a 1-month placebo run-in. Patients were tested at home or work using an interactive computer-telephone system. Standardized measurements of mood, cognitive performance, and food cravings were made before and 30, 90, and 180 minutes after consumption of active and placebo beverages during the late luteal phase of the menstrual cycle. RESULTS: The experimental carbohydrate intervention significantly decreased self-reported depression, anger, confusion, and carbohydrate craving 90-180 minutes after intake. Memory word recognition was also improved significantly compared with scores obtained during the placebo run-in month (P < .05). The isocaloric placebo interventions had no significant effect on any of these measures. CONCLUSION: The results suggest that the psychological and appetitive symptoms of PMS can be relieved after consuming a specially-formulated, carbohydrate-rich beverage known to increase serum tryptophan levels.

Determination of tumor aggressiveness in colorectal cancer by K-ras-2 analysis.

Markers that predict tumor aggressiveness on a case-by-case basis would enable individualization and optimization of oncologic therapy. To achieve this goal, the presence and specific type of K-ras-2 point mutation was determined from formalin-fixed, paraffin-embedded tissue sites in 247 primary and 166 metastatic-recurrent colorectal adenocarcinomas, using a novel approach consisting of topographic tissue selection, DNA amplification, and direct sequencing applicable to large and needle-biopsy-sized specimens. The results provide the basis for a genotypic classification of colorectal cancer capable of predicting individual tumor aggressiveness, including the pattern and extent of metastasis.

K-ras-2 topographic genotyping of pancreatic adenocarcinoma.

OBJECTIVE: To determine the frequency distribution of K-ras-2 point mutation genotypes in pancreatic adenocarcinoma and to evaluate the effectiveness of K-ras-2 genotyping as a means to predict localized disease and potential long-term survival. DESIGN: Topographic genotyping from archival formalin-fixed, paraffin-embedded large- and biopsy-sized tissue specimens as well as cytologic fluid using polymerase chain reaction products and direct sequencing together with clinicopathologic and statistical analysis. SETTING: Tertiary care medical center with molecular diagnostics pathology laboratory. PATIENTS: Patients treated between 1988 and 1993 at Rhode Island Hospital, Providence, yielding 55 primary and 56 metastatic specimens of pancreatic adenocarcinoma. RESULTS: Each primary pancreatic adenocarcinoma was found to contain one of eight specific genotypes that was maintained in all metastatic deposits of that individual tumor. Primary adenocarcinomas confined to the pancreatic bed at diagnosis were predominantly of a normal genotype (56% [14/25]). Pancreatic adenocarcinomas progressing to distant hematogenous metastasis were almost exclusively mutated (88% [7/8]; P < .005). Patients undergoing pancreatic resection (Whipple's operation) and having a normal K-ras-2-genotype (58% [11/19]) had a significantly longer survival (21.3 months) than similar patients with mutated tumors (8.2 months). CONCLUSIONS: The findings support the feasibility of K-ras-2 topographic genotyping to identify potentially indolent disease and suggest a potentially useful role in the preoperative evaluation of pancreatic adenocarcinoma.

Increased expression of complement component 3 in human ectopic endometrium compared with the matched eutopic endometrium.

OBJECTIVE: To compare the gene expression of complement component 3 (C3) in human eutopic and ectopic endometrium. DESIGN: A prospective, controlled study. SETTING: Academic hospital. PATIENT(S): Women with documented endometriosis. INTERVENTION(S): Eutopic and ectopic endometrial tissues were collected simultaneously at laparoscopy. MAIN OUTCOME MEASURE(S): Detection of C3 messenger RNA (mRNA) by in situ hybridization and C3 protein by immunohistochemistry and Western blot. RESULT(S): Expression of C3 mRNA increased in ectopic endometrium compared with that in the matched eutopic endometrium. The quantitative analysis of C3 mRNA by grain count (mean +/- SE) showed 175.60 +/- 40.02 and 39.97 +/- 8.17 grains per micron2 in ectopic and eutopic glands, respectively, and 67.65 +/- 29.82 and 15.02 +/- 5.80 grains per micron2 in ectopic and eutopic stroma, respectively. Expression of C3 mRNA in ectopic glands was significantly higher than that in eutopic glands. The pattern of immunoreactive staining of C3 protein was consistent with that of C3 mRNA. A higher level of C3 protein in ectopic endometrium than eutopic endometrium was detected by immunohistochemistry and Western blot. CONCLUSION(S): Expression of C3 mRNA and protein significantly increased in human ectopic endometrium compared with that in the matched eutopic endometrium.

Preparation by differential gradient centrifugation is better than swim-up in selecting sperm with normal morphology (strict criteria).

OBJECTIVE: To evaluate two commonly used methods of sperm preparation with respect to their effects on sperm morphology (strict criteria). DESIGN: Auto-controlled, split sample study performed on the semen of 74 male partners of couples enrolled for IVF. SETTING: In vitro fertilization and andrology laboratories at a tertiary care, major teaching hospital. PATIENT(S): Seventy-four male partners of couples who were scheduled to undergo IVF. INTERVENTION(S): Equal halves of the same semen sample were evaluated for strict criteria sperm morphology before and after preparation by differential gradient centrifugation using Percoll (Pacific Andrology, Montrose, CA) and by the standard swim-up method. MAIN OUTCOME MEASURE(S): The percentage of morphologically normal sperm was assessed using strict criteria before and after the two methods of sperm preparation. Specific parameters studied were individual abnormalities of the head, midpiece, and tail. RESULT(S): Sperm preparation using differential gradient centrifugation with Percoll produced a significantly greater number of specimens with normal sperm morphology and also showed higher absolute quantitative improvement over the swim-up method. The two methods were comparable in regard to their effects on specific sperm abnormalities (i.e., head, midpiece, and tail defects). CONCLUSION(S): The differential gradient sperm separation method using Percoll is superior to the swim-up method for selecting sperm with normal morphology as assessed by strict criteria. Because sperm morphology as assessed by strict criteria is a good predictor of oocyte fertilization, this method can be recommended as the method of choice for assisted reproductive technology laboratories. Use of this method may help improve outcome by increasing fertilization rates.

Elevated expression of the proapoptotic BCL-2 family member, BAK, in the human endometrium coincident with apoptosis during the secretory phase of the cycle.

OBJECTIVE: To characterize the distribution of BAK (BCL-2 homologous antagonist/killer) protein in the human endometrium relative to the occurrence of apoptosis. DESIGN: A prospective, controlled study. SETTING: An academic hospital. PATIENT(S): Premenopausal women with histologically normal endometrium who were undergoing hysterectomy and curettage. INTERVENTION(S): Endometrial tissues were collected. MAIN OUTCOME MEASURE(S): Detection of BAK protein by immunohistochemical and immunoblot analyses, and of apoptosis by in situ DNA end-labeling. RESULT(S): BAK protein was detected in secretory endometrium and was confined to the glandular epithelial cells of the functionalis layer. Immunoreactive BAK was absent from most of the cells of the proliferative endometrium. By immunoblot analysis, we confirmed the immunohistochemical data by demonstrating that a 30-kD protein corresponding to BAK was elevated in lysates prepared from secretory, as compared with proliferative, endometrium. The elevated levels of BAK coincided with the onset of apoptosis in endometrial glandular epithelial cells during the secretory phase of the menstrual cycle. CONCLUSION(S): BAK protein localizes to glandular epithelial cells on the verge of apoptosis in the human endometrium. Thus, BAK likely functions with other members of the BCL-2 family in the regulation of apoptosis in the human uterus.

Immunohistochemical localization of alpha-2 macroglobulin receptor/low-density lipoprotein receptor-related protein, receptor-associated protein, and Gp330 in the human endometrium.

OBJECTIVE: We investigated the expression of alpha-2 macroglobulin receptor/low-density lipoprotein receptor-related protein (LRP), an analogous receptor Gp330, and the 39-kd receptor-associated protein (RAP) in the human endometrium. METHODS: Specific polyclonal and monoclonal antibodies against LRP, Gp330, and RAP were used in standard immunohistochemical analyses of normal secretory and proliferative archival endometrial tissue. RESULTS: There was prominent and uniform stromal staining for LRP in secretory and proliferative endometrium. In both phases, 10-25% of glands stained positive for Gp330, with no appreciable stromal staining for Gp330. Also in both phases, 15-30% of glands stained positive for RAP, with apical staining in proliferative glands and uniform staining in secretory glands. Stromal staining for RAP was patchy and appeared to be more intense in the secretory samples than in the proliferative ones. CONCLUSIONS: The human endometrium expresses LRP, RAP, and Gp330. These receptor proteins are known to be involved in endocytosis of multiple ligands. Some of these ligands, such as proteases, plasminogen activators, and cytokines, are produced by the endometrium and play a role in endometrial remodeling and receptivity to implantation. By virtue of their in vitro properties, it is conceivable that endometrial LRP, Gp330, and RAP are involved in endometrial physiology.

Late recurrence of ductal carcinoma in situ at the cutaneous end of surgical drainage following total mastectomy.

A 40-year-old woman underwent upper outer quadrantectomy breast biopsy which revealed extensive intraductal carcinoma, predominantly comedocarcinoma type, with high nuclear grade. Involved ducts were transected at the edge of the biopsy. Total mastectomy with low axillary lymph node dissection was performed 2 weeks later, showing residual intraductal carcinoma in the upper inner quadrant and no evidence of metastasis. Eight years later, the patient developed two separate foci of recurrent, invasive ductal carcinoma at the exit sites for mastectomy drainage in the subcutaneous skin of the upper abdomen. The mastectomy scar was clinically free of tumor. The biological basis for this unusual sequela of treated intraductal carcinoma is discussed together with its importance for management of early breast cancer.

Perinatal outcome of singleton pregnancies conceived by in vitro fertilization: a controlled study.

To determine whether conception by in vitro fertilization and embryo transfer (IVF) predisposes to perinatal complications, the obstetric records of 54 women delivered of singleton pregnancies after conception by IVF were examined. Control women were matched for age, parity, race, year of delivery, diethylstilbestrol exposure and medical problems; another group of women who conceived after infertility treatment was matched in similar fashion. IVF patients showed a longer first stage of labor than previously infertile women, experienced a greater intrapartum blood loss than control or previously infertile women, and showed a trend toward a higher cesarean delivery rate than control women. The differences noted probably do not arise from the physiology of IVF, and although some differences are statistically significant, they are of minimal clinical significance. Singleton pregnancies arising after IVF should not be considered as high risk in the absence of other predisposing factors.

Rupture of the renal pelvis associated with intravenous fluid bolus.

Previously reported cases of spontaneous rupture of the kidney or ureter reflect underlying renal pathology and have been reported both spontaneously and in relationship to delivery, but not in relation to other operative procedures. A 27-year-old woman at 19 weeks' gestation developed severe right flank pain in the operating room immediately prior to cerclage placement. Postoperative renal ultrasound examination and intravenous pyelogram performed to evaluate persistent flank pain demonstrated renal pelvis rupture. No other renal abnormality was present. We postulated that increased urine flow from the fluid bolus for the spinal anesthetic precipitated the rupture.

Ovarian function after conservational ovarian surgery: a long-term follow-up study.

OBJECTIVE: To determine the long-term effects of conservational ovarian surgery on subsequent ovarian function. METHOD: Medical, surgical and menstrual records of 87 reproductive age women who have undergone pelvic surgery for indications including endometriosis, infertility, and pelvic pain between June 1982 and June 1989 were retrospectively reviewed. RESULTS: Sixty-seven patients had ovarian surgery during their procedure; 26 ovarian cystectomies (OC), 19 partial ovarian resections (OR) and 22 ovariolyses (OL). Twenty patients had no ovarian surgery (NOS). The mean follow-up period was comparable in all groups (OL 32 +/- 15 months, OR 34 +/- 26 months, OC 41 +/- 30 months and NOS 37 +/- 21 months). The mean time to onset of the first postoperative menses was not significantly different among the four groups (OL 24 +/- 7 days, OR 24 +/- 13 days, OC 29 +/- 26 days and NOS 22 +/- 6 days). The cumulative conception rates were not significantly different (OL 23%, OR 37%, OC 19% and NOS 25%). Menstrual disturbances, defined as perceived deviations from preoperative patterns, did not appear to be related to the type or extent of ovarian surgery and occurred in comparable frequencies among patients with or without ovarian surgery (OL 23%, OR 37%, OC 23% and NOS 20%). In the majority of cases, menstrual disturbances occurred either in the early postoperative months and were self-limited, or much later in which case they were related to recurrent endometriosis. Premature ovarian failure has so far occurred in one patient suggesting an incidence comparable to that in the general population. CONCLUSIONS: It is concluded that in this group of patients, conservational ovarian surgery had no significant effects on ovulatory and menstrual function over a prolonged follow-up period.