Perioperative morbidity of different operative approaches in early cervical carcinoma: a systematic review and meta-analysis comparing minimally invasive versus open radical hysterectomy.

PURPOSE: Radical hysterectomy and pelvic lymphadenectomy is the standard treatment for early cervical cancer. Studies have shown superior oncological outcome for open versus minimal invasive surgery, but peri- and postoperative complication rates were shown vice versa. This meta-analysis evaluates the peri- and postoperative morbidities and complications of robotic and laparoscopic radical hysterectomy compared to open surgery. METHODS: Embase and Ovid-Medline databases were systematically searched in June 2020 for studies comparing robotic, laparoscopic and open radical hysterectomy. There was no limitation in publication year. Inclusion criteria were set analogue to the LACC trial. Subgroup analyses were performed regarding the operative technique, the study design and the date of publication for the endpoints intra- and postoperative morbidity, estimated blood loss, hospital stay and operation time. RESULTS: 27 studies fulfilled the inclusion criteria. Five prospective, randomized-control trials were included. Meta-analysis showed no significant difference between robotic radical hysterectomy (RH) and laparoscopic hysterectomy (LH) concerning intra- and perioperative complications. Operation time was longer in both RH (mean difference 44.79 min [95% CI 38.16; 51.42]), and LH (mean difference 20.96 min; [95% CI - 1.30; 43.22]) than in open hysterectomy (AH) but did not lead to a rise of intra- and postoperative complications. Intraoperative morbidity was lower in LH than in AH (RR 0.90 [0.80; 1.02]) as well as in RH compared to AH (0.54 [0.33; 0.88]). Intraoperative morbidity showed no difference between LH and RH (RR 1.29 [0.23; 7.29]). Postoperative morbidity was not different in any approach. Estimated blood loss was lower in both LH (mean difference - 114.34 [- 122.97; - 105.71]) and RH (mean difference - 287.14 [- 392.99; - 181.28]) compared to AH, respectively. Duration of hospital stay was shorter for LH (mean difference - 3.06 [- 3.28; - 2.83]) and RH (mean difference - 3.77 [- 5.10; - 2.44]) compared to AH. CONCLUSION: Minimally invasive radical hysterectomy appears to be associated with reduced intraoperative morbidity and blood loss and improved reconvalescence after surgery. Besides oncological and surgical factors these results should be considered when counseling patients for radical hysterectomy and underscore the need for new randomized trials.

Protective operative techniques in radical hysterectomy in early cervical carcinoma and their influence on disease-free and overall survival: a systematic review and meta-analysis of risk groups.

PURPOSE: Radical hysterectomy with pelvic lymphadenectomy presents the standard treatment for early cervical cancer. Recently, studies have shown a superior oncological outcome for open versus minimal invasive surgery, however, the reasons remain to be speculated. This meta-analysis evaluates the outcomes of robotic and laparoscopic hysterectomy compared to open hysterectomy. Risk groups including the use of uterine manipulators or colpotomy were created. METHODS: Ovid-Medline and Embase databases were systematically searched in June 2020. No limitation in date of publication or country was made. Subgroup analyses were performed regarding the surgical approach and the endpoints OS and DFS. RESULTS: 30 studies fulfilled the inclusion criteria. Five prospective, randomized-control trials were included. Patients were analyzed concerning the surgical approach [open surgery (AH), laparoscopic surgery (LH), robotic surgery (RH)]. Additionally, three subgroups were created from the LH group: the LH high-risk group (manipulator), intermediate-risk group (no manipulator, intracorporal colpotomy) and LH low-risk group (no manipulator, vaginal colpotomy). Regarding OS, the meta-analysis showed inferiority of LH in total over AH (0.97 [0.96; 0.98]). The OS was significantly higher in LH low risk (0.96 [0.94; 0.98) compared to LH intermediate risk (0.93 [0.91; 0.94]). OS rates were comparable in AH and LH Low-risk group. DFS was higher in the AH group compared to the LH group in general (0.92 [95%-CI 0.88; 0.95] vs. 0.87 [0.82; 0.91]), whereas the application of protective measures (no uterine manipulator in combination with vaginal colpotomy) was associated with increased DFS in laparoscopy (0.91 [0.91; 0.95]). CONCLUSION: DFS and OS in laparoscopy appear to be depending on surgical technique. Protective operating techniques in laparoscopy result in improved minimal invasive survival.

Morbidity, fertility and pregnancy outcomes after myoma enucleation by laparoscopy versus laparotomy.

OBJECTIVES: Myomas are defined as benign tumours that arise from smooth muscle cells of the uterus. Clinically, they are found in 5-77% of women of reproductive age. The prevalence rate varies considerably in the literature and a large number of fibroids do not cause symptoms. The lifetime risk of acquiring myomas is 70% for Caucasian women and ≥ 80% for African American women. MATERIALS/METHODS: The data of 265 patients undergoing surgery for symptomatic myomas by laparoscopy or laparotomy, performed in the gynaecological department of Hannover Medical School, Hannover, Germany, between 2009 and 2013, were retrospectively analysed in this retrospective design study. RESULTS: High pregnancy rates (up to 70%) and birth rates (up to 86%) after myomectomy, regardless of the surgical approach adopted, were found in the current study. The trend was that ≥ 3 myomas and those that were ≥ 6 cm in size were almost always removed by laparotomy in our clinic. It was possible to remove up to 42 myomas without having to perform a hysterectomy. A statistically significant negative correlation was observed in relation to the association between the size of the largest myoma extracted and the pregnancy rate (p = 0.02). A statistically significant correlation between the number of removed myomas and the pregnancy rate was observed for patients who wished to bear children (p = 0.010). Elevated complication rates (of up to 50%) were reported for more than three extracted myomas with a statistically significance (p = 0.0471). CONCLUSIONS: It is necessary to ensure sound preoperative selection of the surgical approach in order to achieve the most optimal results, especially for those patients who wished to bear children.

SPECT/CT for SLN dissection in vulvar cancer: Improved SLN detection and dissection by preoperative three-dimensional anatomical localisation.

OBJECTIVE: In this study, we assessed the feasibility and clinical advantages of single photon emission computed tomography with CT (SPECT/CT) for sentinel lymph node (SLN) detection in vulvar cancer. METHODS: This is a unicentric prospective trial. Vulvar cancer patients underwent preoperative SLN marking (10MBq Technetium (TC)-99m-nanocolloid) and subsequent planar lymphoscintigraphy (LSG) and SPECT/CT for SLN visualization. Directly before surgery, a patent blue dye was injected. We assessed detection rates of SPECT/CT and those of planar LSG and intraoperative detection. We analyzed the sensitivity, negative predictive value and false negative rate. RESULTS: At Hannover Medical School, 40 vulvar cancer patients underwent SLN dissection after preoperative LSG and SPECT/CT. The mean diameter of all tumors in final histology was 2.23 (0.1-10.5) cm with a mean tissue infiltration of 3.93 (0.25-11) mm. In preoperative imaging, SPECT/CT identified significantly more SLNs (mean 8.7 (1-35) LNs per patient) compared to LSG (mean 5.9 (0-22) LNs, p<0.01). In addition, SPECT/CT led to a high spatial resolution and anatomical localization of SLNs. Thus, SPECT/CT identified aberrant lymphatic drainage in 7/40 (17.5%) patients. There were no significant differences, but significant correlation was found between SPECT/CT and intraoperative SLN identification. Regarding inguino-femoral LNs, for all patients who underwent complete groin dissection, sensitivity was 100%, NPV was 100% and false negative rate was 0%. CONCLUSION: SPECT/CT leads to higher SLN identification compared to LSG in vulvar cancer. Due to its higher spatial resolution and three-dimensional anatomical localisation of SLNs, SPECT/CT provides the surgeon with important additional information, facilitates intraoperative SLN detection and predicts aberrant lymphatic drainage.

Isthmocervical labelling and SPECT/CT for optimized sentinel detection in endometrial cancer: technique, experience and results.

OBJECTIVE: We evaluated the clinical feasibility of a new injection technique for sentinel detection in endometrial carcinoma (EC), transcervical subepithelial injection into the isthmocervical region of the myometrium. We compared detection of sentinel lymph nodes (SLN) by single photon emission computed tomography with CT (SPECT/CT) with planar lymphoscintigraphy. METHODS: This is a unicentric prospective study. In all patients, transcervical injection of 10 MBq Technetium-99m-nanocolloid was performed into the isthmocervical myometrium without anaesthesia. After 40 (30-60) min, lymphoscintigraphy and SPECT/CT were performed. Patent blue was administered before surgery. The number and localisation of SLN detected in SPECT/CT and lymphoscintigraphy were recorded and compared to the SLN and non-SLN dissected intra-operatively. RESULTS: Between August 2008 and March 2012, 31 patients with EC were enrolled. The new transcervical injection of labelling substances led to high intra-operative (90.3%) detection rates, pelvic bilateral (57%), para-aortic (25%). SPECT/CT significantly identified more SLN than lymphoscintigraphy (mean 2.2 (1-8) to 1.3 (1-7)) in more patients (29/31 (93.5%) to 21/31 (68%), p<0.01). If SLN were identified in one hemi-pelvis, the histological evaluation of the SLN correctly predicted lymph node (LN) metastases for this basin which led to sensitivity 100%, negative predictive value (NPV) 100%, and false negative results 0%. CONCLUSION: Transcervical SLN marking in combination with SPECT/CT is easily applicable and leads to high physiologic detection rates in pelvic and para-aortic lymphatic drainage areas. Non-affected SLN truly predicted a non-affected LN basin. Combining both methods SLN dissection may be a safe and feasible staging technique for clinical routine in EC.

Use of the Sentinel Lymph Node Technique Compared to Complete Inguino-femoral Lymph Node Removal in Patients with Invasive Vulvar Cancer in Germany.

In the current S2 guidelines, the standard surgical therapy for patients with vulvar cancer also includes inguino-femoral lymphadenectomy. However, in view of the severe side-effects associated with this approach such as problems with wound healing, lymphoceles and lymphoedema, the search is on for alternative treatments that could decrease treatment-associated morbidity and improve patients' quality of life, particularly for node-negative patients. The sentinel lymph node technique is currently the gold standard in the treatment of unifocal breast cancer (clinically negative axilla), and studies on the use of this technique in the treatment of vulvar cancer are promising. To date, the diagnostic accuracy of this method in vulvar cancer has only been evaluated in a single, one-arm, non-randomised, multicentre study. In preparation for a multicentre study, in 2010 we surveyed 41 German hospitals to investigate how often they used the sentinel lymph node technique compared to inguino-femoral lymphadenectomy. The hospitals were grouped according to hospital size and number of patients treated for vulvar cancer. The decision criteria to determine the type of procedure performed were also investigated. Finally, the hospitals were asked whether they would be willing to participate in a prospective clinical study to evaluate the sentinel lymph node technique in patients with vulvar cancer. The majority of surgeons questioned (73 %) already had some experience with this technique in patients with vulvar cancer. In our survey, 27 % of hospitals carried out inguino-femoral lymphadenectomy, 10 % used the sentinel lymph node technique, and 63 % used both methods. In 24 % of hospitals, the standard procedure consisted of the sentinel lymph node technique supplemented by inguino-femoral lymphadenectomy. Only 20 % of the institutions surveyed in our study carried out sentinel lymph node biopsy alone in accordance with the criteria of the consensus recommendations. The majority of the investigated institutions were willing to participate in a randomised prospective clinical study to evaluate the effectiveness of sentinel lymph node sampling in patients with vulvar cancer.

Patent blue sentinel node mapping in cervical cancer patients may lead to decreased pulse oximeter readings and positive methaemoglobin results.

BACKGROUND AND OBJECTIVE: Patent blue (4-[(4-diethylaminophenyl)-(4-diethylazaniumylidencyclohexa-2,5-dienyliden) methyl]-6-hydroxy-3-sulfo-benzolsulfonate, sodium salt) is a contrast dye used for the intraoperative detection of the primary lymphatic nodes draining the area of tumour infiltration. The dye is known to interact with pulse oximeter readings. However, the degree of alteration seems to be moderate and predictable when patent blue is injected into the perimammilar region during breast surgery. METHODS: Here we report severe interference with the anaesthetic monitoring when patent blue was injected into the cervix prior to laparoscopy-assisted radical vaginal hysterectomy for cervical cancer. RESULTS: Injection of patent blue into the cervix induced a rapid (within 14 +/- 9 min after the injection) and severe (from 98% to 89 +/- 2%) decrease in pulse oximeter readings, accompanied by positive methaemoglobin values of 7.3 +/- 2.5% (arterial co-oximetry, Bayer Rapidlab 865 blood gas analyser; Bayer, Fernwald, Germany). Control of these values by a different device (Radiometer ABL co-oximeter blood gas analyser; Radiometer, Willich, Germany) yielded negative methaemoglobin results (<1.7%, mean 0.9 +/- 0.6%). The arterial PO2 was normal in all patients throughout the procedure. CONCLUSION: Injection of patent blue into the cervix uteri interferes dramatically with pulse oximeter readings. This situation is further complicated by device-dependent arterial co-oximetry methaemoglobin results. For the time being it is recommendable to monitor adequate oxygenation of the patient in the presence of patent blue by regular control of the arterial PO2. Clearly, the unresolved issue of reliable methaemoglobin determination in the presence of patent blue remains a matter of clinical concern for anaesthetists.

Dilution of dye improves parametrial SLN detection in patients with cervical cancer.

OBJECTIVE: Aim of this study to improve the detection rate of parametrial sentinel nodes in patients with early cervical cancer by using a new dosage of blue dye in a randomized trial. METHODS: Standard labelling volume of 4 ml Patent Blue was compared to 2 ml Patent Blue diluted with 8 ml NaCl 0.9% in 60 patients using a randomized protocol. Tc-99 was not applied in any patient. All patients underwent open lymphadenectomy. RESULTS: In each arm 30 patients were enrolled. Overall detection rate of sentinel nodes was 93.3%. Both groups did not differ with regard to patient's age, BMI, tumor stage, number of lymph nodes harvested, number of sentinel nodes detected and detection rate. Significantly more patients with parametrial sentinel nodes were detected in the diluted protocol (37.0% vs. 10.3%) which is due to the lateral part of the cardinal ligament (29.6% vs. 6.9%). CONCLUSION: In our study overall detection of sentinel nodes using a dye was high. The diluted solution of Patent Blue led to a higher detection rate of parametrial sentinel nodes. These findings might help explain the discrepancy between the high reported rate of parametrial lymph nodes in anatomical studies compared with the low rates in clinical sentinel series.

[Perinatal morbidity in pregnancies of women with preconceptional and gestational diabetes mellitus in comparison with pregnancies of non-diabetic women. Results of the perinatal registry of Lower Saxony, Germany]. / Perinatale Morbidität in Schwangerschaften von präkonzeptionellen Diabetikerinnen und Gestationsdiabetikerinnen im Vergleich mit Nichtdiabetikerinnen. Ergebnisse der Niedersächsischen Perinatalerhebung.

OBJECTIVE: Pregnancies of women with glucose tolerance disorders are still associated with high maternal and perinatal morbidity. In the present population-based study, we investigated the fetal and neonatal risks and complications in pregnancies of women with preconceptional and gestational diabetes in comparison with pregnancies of non-diabetic women. METHODS: A total of 913,255 pregnancies with 926,685 newborns documented by the Perinatal Registry of Lower Saxony, Germany, between 1987 and 1999 was retrospectively evaluated. In particular, the obstetric parameters of 3439 women with preconceptional diabetes mellitus (PDM, n = 3485 neonates), 4051 women with gestational diabetes mellitus (GDM, n = 4155 neonates) and 905,765 non-diabetic women (NDM, n = 919.053 neonates) were analysed. RESULTS: The diabetes groups had a higher rate of preterm birth < 31st week of gestation (PDM vs. NDM odds ratio [OR] 2.341; GDM vs. NDM OR 1.483) and between the 32nd and 37th weeks of gestation (OR 3.688 and 2.418, respectively). The incidences of pathological cardiotocograms (OR 1.519 and 1.258, respectively) and acidosis as indication for operative delivery (OR 5,015 and 2,102, respectively) were increased in the diabetes groups. There was a higher prevalence of birth weight > 4500 g (OR 2.775 and 2.742, respectively) and >95. percentile (OR 4.268 and 3.610, respectively) in newborns of diabetic women. The higher rate of umbilical arterial pH < 7.00 (OR 2.481 and 1.195, respectively) as well as Apgar score at 1 minute <7 (OR 2.971 and 1.897, respectively) indicated a poorer neonatal outcome in pregnancies of women with diabetes mellitus. The frequency of necessary intubation of the infants was significantly higher in the diabetes group (OR 3.384 and 1.317, respectively). There also was an increased prevalence of intrauterine fetal death in pregnancies of diabetic women (OR 4.197 and 2.511, respectively). CONCLUSION: Pregnancies of women with diabetes mellitus are still correlated with higher perinatal risks and complications. The neonatal morbidity in pregnancies with gestational diabetes mellitus was more similar to that of pregnancies with preconceptional diabetes mellitus than to the morbidity in the group of non-diabetic women. Changes of the parameters within the observation period were the same in all three groups and therefore cannot be attributed to a primary more intensive prenatal care of diabetic women.

Is the decision for colorectal resection justified by histopathologic findings: a prospective study of 100 patients with advanced ovarian cancer.

OBJECTIVE: We compare the indication for colorectal resection in patients with advanced ovarian cancer with histopathologic findings. We describe the effect on pelvic control and morbidity associated with surgery. METHODS: Between February 1995 and March 2001, 100 patients with FIGO stage IIIc ovarian cancer underwent pelvic en bloc resection with excision of the rectosigmoid colon as part of primary or secondary cytoreductive surgery. Decision for resection was made by the surgeon when tumor involvement of the cul-de-sac was suspected. Rectosigmoid infiltration was histopathologically defined as infiltration of the serosa or deeper. RESULTS: In 73 of 100 patients (73%) tumor involvement of the rectum was confirmed histopathologically: infiltration of the serosa in 28 (28%) patients, infiltration of the muscularis in 31 (31%) patients, and infiltration of the mucosa in 14 (14%) patients; in 27 (27%) patients no infiltration was found. Histopathologically confirmed pelvic R0 resection was achieved in 85 (85%) patients. In 11 (11%) patients the pelvic resection margins were tumor-involved and in four (4%) patients visible parametric tumor remained in situ. Pelvic recurrence occurred in 4 (4.7%) of 85 optimally debulked patients compared with 9 (60%) of 15 patients with suboptimal pelvic resection status (P < 0.05). End colostomy could be prevented in 94 (94%) of 100 patients. CONCLUSION: Pelvic en bloc surgery with rectosigmoid resection was justified by histopathologic outcome since deperitonealization with preservation of the rectosigmoid would have left tumor in situ in 73% of patients with suspected cul-de-sac involvement.

Repellent and toxic effects of plant extracts on subterranean termites (Isoptera: Rhinotermitidae).

The effects of four plant extract formulations on the orientation and survival of subterranean termites were tested. In choice experiments, extract-treated filter paper had a significantly repellent effect on groups of Reticulitermes santonensis De Feytaud, R. virginicus (Banks), Coptotermes formosanus Shiraki, and Schedorhinotermes intermedius Breinli. There was no species-specific difference in avoidance behavior toward the tested concentrations. No-choice experiments revealed toxic properties of all investigated extracts by contact or airborne compounds against R. santonensis. However, high termite mortality was only achieved by forced direct or forced indirect exposure to the plant material, Feeding deterrence or digestive toxicity could not be judged in these experiments. One of the extracts was efficiently used for soil treatments to protect a food substrate against R. santonenis infestation. Extract-treated barriers in the experiments did not affect mortality compared with control trials but prevented termites from penetrating treated soil.

Trocar-site metastasis is not always due to laparoscopy.

The use of laparoscopic surgical techniques for the management of gynecologic malignancies has increased over the last years. Metastasis developing at the trocar insertion site is an emerging problem. We present the case of a 66-year-old woman with endometrial cancer who was diagnosed with an umbilical tumor after laparoscopically assisted vaginal hysterectomy (LAVH) and bilateral salpingoophorectomy. The interval between LAVH and diagnosis of the umbilical tumor was 13 months. The tumor was excised, and metastasis of endometrial cancer was histologically confirmed. Review of computer tomograms taken before LAVH showed a tumor in the umbilical area that had not been recognized before therapy. Therefore, tumor manifestation at the abdominal wall after laparoscopic surgery should not automatically be considered the result of iatrogenic spreading.

Laparoscopic lymph node staging of cervical cancer in the 19th week of pregnancy. A case report.

We present the case of a 39-year-old gravida I para 0 woman who underwent laparoscopic staging of lymph node involvement in cervical cancer in the 19th week of pregnancy. She had been diagnosed with adenosquamous carcinoma of the cervix, stage 1B1, grade 2, with tumor involvement of the lymphovascular space and tumor involved resection margins via a cone biopsy in the 16th week of pregnancy. In order to decide whether it would be safe to proceed with the pregnancy, she was submitted to the laparoscopic exposure and removal of 18 parametric and pelvic lymph nodes. One positive lymph node was detected at the right internal iliac artery; therefore, an open radical hysterectomy with paraaortic lymphadenectomy was performed. This case shows that lymph node staging for cervical cancer can be done laparoscopically in the 2nd trimester. Information yielded during the course of this procedure can be crucial in deciding whether it is possible to preserve the pregnancy.

Human cardiac microvascular and macrovascular endothelial cells respond differently to oxidatively modified LDL.

Oxidation of low density lipoproteins (LDL) is considered a key event in the pathogenesis of atherosclerotic lesions. Disturbed generation of coagulatory and anticoagulatory factors by endothelial cells contributes to thrombosis and the progression of atherosclerosis in coronary arteries. In this study, the effects of native LDL (n-LDL) and oxidized LDL (ox-LDL) on human coronary endothelial cells were measured. The reaction of coronary endothelial cells to LDL were compared with those of cardiac microvascular endothelial cells grown under comparable conditions. LDL was isolated by ultracentrifugation and copper oxidized. The degree of oxidation was expressed as malondialdehyd (MDA) equivalents and was 0.78+/-0.14 nM MDA/mg LDL for native LDL and 13.63+/-1.18 nmol MDA/mg LDL for ox-LDL. Basal secretion of t-PA and PAI-1 activity were higher in macrovascular endothelial cells. Incubation of n-LDL in concentrations ranging from 3 to 100 microM/ml LDL-protein did not change t-PA-secretion, PAI-1 activity or procoagulant activity in both cell types. Ox-LDL (3 to 100 microM/ml LDL protein) decreased t-PA secretion in a concentration dependent manner from 30.9+/-1.7 to 13.7+/-30 ng/ml per 24 h per 10(6) cells (P < 0.01), increased PAI-1 antigen from 2772+/-587 to 4441+/-766 ng/ml per 24 h per 10(6) cells (P < 0.05) as well as PAI-1 activity from 34+/-6 to 55+/-9 AU/ml per 24 h per 10(6) cells (P < 0.05) in macrovascular endothelial cells but had only minor effects on microvascular endothelial cells. Procoagulant activity measured as coagulation time, similarly increased only in macrovascular endothelial cells from 197+/-6 to 76+/-6 s/24 h per 10(6) cells (P < 0.05). The effect on PAI-1 secretion showed a dependency to the degree of oxidation and could be completely blocked by the antioxidant probucol. The angiotensin converting enzyme (ACE), which represents an endothelial enzyme not related to coagulation, remained unchanged during incubation with ox-LDL. Basal ACE activity was higher in microvascular endothelial cells. The higher susceptibility of macrovascular endothelial cells to ox-LDL may partially determine the localization of thrombus formation and the development of atherosclerotic plaques in hyperlipidemic patients.

Hemicellulose-degrading bacteria and yeasts from the termite gut.

Termites play a major role in the recycling of photosynthetically fixed carbon. With the aid of their symbiotic intestinal flora, they are able to degrade extensively wood constituents such as cellulose and hemicellulose. Nevertheless, the microbial species involved in the degradation of hemicelluloses are poorly defined. The purpose of this paper was to examine the microflora involved in hemicellulose degradation. Different aerobic and facultatively anaerobic bacteria and yeasts were isolated using xylan, arabinogalactan and carboxymethylcellulose as substrates. Gram-positive isolates belonged to the genera Bacillus, Paenibacillus, Streptomyces or the actinobacteria group, while the Gram-negative strains were assigned to the genera Pseudomonas, Acinetobacter, Ochrobactrum, and to genera belonging to the family Enterobacteriaceae. The spectrum and activity of xylan- and arabinogalactan-hydrolysing glycosidases of these new isolates, together with additional bacterial strains originally obtained from enrichments with aromatic compounds were determined.

Isolation and characterization of macrovascular and microvascular endothelial cells from human hearts.

In vivo models to investigate mechanisms of local hemostasis in the macro- and microvascular coronary circulation are not available. Therefore, we established a culture system of human macro- and microvascular endothelial cells with high cellular yield and high endothelial cell purity. Microvascular endothelial cells from human hearts were isolated by enzymatic treatment of cardiac muscle preparations obtained during heart transplantation. The isolated microvessels were used to start cultures that were subsequently separated and purified from contaminating nonendothelial cells by paramagnetic beads linked to the lectin Ulex europaeus agglutinin I. Macrovascular endothelial cells were isolated from epicardial coronary arteries and purified by paramagnetic beads as well. With this method high purity (< 2% nonendothelial cells) was achieved as judged from fluorescence-activated cell sorting. Immunochemistry demonstrated the expression of several typical endothelial markers. The two endothelial cell types displayed functional heterogeneity in respect to bradykinin degradation and plasminogen activator inhibitor-1 activity. Thus the ability to selectively isolate and culture human macro- and microvascular cardiac endothelial cells provides a valuable tool to systematically investigate endothelial function in human hearts.

Cultivation and characterization of micro- and macrovascular endothelial cells from the human heart.

In order to investigate processes, such as atherosclerosis and inflammation in vitro, it is necessary to obtain viable and pure endothelial cell cultures from human hearts. To this end, endothelial cells were isolated and cultured from the micro- and macrovasculature of human hearts obtained during heart transplantation. Isolation of capillaries after enzymatic digestion of heart muscle provided a source of microvascular endothelial cells. Contaminating non-endothelial cells were removed by a new technique: paramagnetic beads linked to the lectin ulex europaeus I (UEA-I) were used to select endothelial cells. The resulting cultures contained less than 2% of non-endothelial cells, as judged from immunological staining and fluorescence-activated cell sorting. Both types of endothelial cell displayed typical endothelial properties. They were all positive for factor VIII-related antigen and expressed the endothelial-specific adhesion molecules, CD31 and E-selectin (ELAM-1), after stimulation with cytokines. In addition, they could be labelled with Dil-Ac-LDL, contained angiotensin converting enzyme activity and secreted tissue plasminogen activator, thus demonstrating that typical endothelial functions were preserved in culture.

Isolation and ultrastructural features of a new strain of Dimastigella trypaniformis Sandon 1928 (Bodonina, Kinetoplastida) and comparison with a previously isolated strain.

A strain of the free-living flagellate Dimastigella trypaniformis Sandon was isolated from the gut contents of the Australian termite Mastotermes darwiniensis Froggatt. The fine structure of the isolate (Ulm strain) was compared with a strain isolated from soil in Scotland (Glasgow strain). The biflagellated trophozoites possess great morphological similarities with the bodonids and cryptobiids such as a polykinetoplastic mitochondrial complex, a flagellar paraxial rod, and a cytopharyngeal complex. However, characteristic features are also found such as the two opposed basal bodies and the two flagella, one anteriorly and the other recurrently directed, the latter adhering to a ventral furrow, a rostrum as well as a cytostome and a cytopharynx with rostral structures. These organisms have a typical motion with a highly flexible cell body. They are able to form resistant cysts in the culture. The two strains are distinguishable by small differences in their growth characteristics in different media and qualitative ultrastructural features.

Opponent colour coding is a universal strategy to evaluate the photoreceptor inputs in Hymenoptera.

Behavioural tests were carried out with 9 hymenopteran insect species, which ranked certain sets of coloured stimuli according to their subjective similarity to a previously memorized stimulus. Kendall's tau co-efficient is employed for the analysis of correlation between these similarity rankings and the colour distance rankings predicted by various models of neural colour computation. The models are based on the measured spectral sensitivities of photoreceptor colour types and use a variety of simple colour coding systems to derive hypothetical colour distances. The correlation between the predictions of the models and the behavioural results serves as a measure for the likelihood of existence of a colour coding system. In all species, the similarity rankings can be best explained by assuming that colour is coded on a perceptual level by two colour opponent mechanisms. Brightness differences are ignored, indicating that an intensity-coding sub-system is not used in colour discrimination by the insects investigated. The weighting factors of the colour opponent mechanisms differ between species in detail, but not in the principles involved. It is thus possible to employ a standard measure of perceptual colour distance (colour hexagon distance) to predict the capacities of colour discrimination adequately in all the tested insects.