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Funder: Università degli Studi di Padova, Evaluating the effects of neonicotinoids on forager bees in conditions as near as possible to those in nature presents a considerable challenge. Tackling this challenge is, however, necessary to establish their negative side effects on these pollinators. For instance, it is still under debate the mechanism by which bees seem to recognize low-level contaminations of neonicotinoid insecticides in nectar and pollen of the flowers they visit and limit collection to protect themselves and their hive from a possible intoxication. In this study, we propose an experimental system that involves the use of foragers in free flight foraging repeatedly on artificial feeders containing a sucrose solution contaminated with clothianidin, as well as foragers feeding at adjacent control feeders, allowing us to observe changes in their foraging activity. The progressive disappearance of foragers from the contaminated feeders became increasingly clear and rapid with the increase in clothianidin concentration. The lowest concentration at which we observed an effect was around 10 µg/L, which corresponds to the maximum residual concentration (10 ng/g) observed in pollen and nectar of flowers close to open fields sown with seeds coated with insecticides. At the highest concentrations tested (80 µg/L), there was an almost total abandonment of the feeders. The estimated quantity of contaminated sucrose solution collected by foragers showed an almost linear relationship inversely proportional to clothianidin concentration, whilst the estimated quantity of insecticide collected by a forager increased and then stabilised at the highest concentrations tested of 40 and 80 µg/L. Irregular mortality was not observed in front of the hives, furthermore, foragers did not show evident memory of the position of the treated units in the trials on the 2 consecutive days. The decrease in foraging activity in the presence of a few µg/L of insecticide in the sucrose solution appears to limit the introduction of elevated amounts of toxic substances into the hives, which would have serious consequences for the young bees and the brood. At the same time, in the absence of an alternative energy source, even reduced feeding of the hive can compromise colony health. more...

Three-dimensional (3D) digital brain atlases and high-throughput brain-wide imaging techniques generate large multidimensional datasets that can be registered to a common reference frame. Generating insights from such datasets depends critically on visualization and interactive data exploration, but this a challenging task. Currently available software is dedicated to single atlases, model species or data types, and generating 3D renderings that merge anatomically registered data from diverse sources requires extensive development and programming skills. Here, we present brainrender: an open-source Python package for interactive visualization of multidimensional datasets registered to brain atlases. Brainrender facilitates the creation of complex renderings with different data types in the same visualization and enables seamless use of different atlas sources. High-quality visualizations can be used interactively and exported as high-resolution figures and animated videos. By facilitating the visualization of anatomically registered data, brainrender should accelerate the analysis, interpretation, and dissemination of brain-wide multidimensional data. more...

The academic world initially regarded Wikipedia with misgivings for a number of reasons, including the uncertainty concerning the reliability of its content, its anonymous authorship and the fact that students often use it as an easy way to cut and paste material for their coursework. In recent years, however, university instructors’ perception of Wikipedia has changed significantly: it is seen as a useful teaching resource, as well as a promising environment for learning and collaborative knowledge building. It can be used for teaching purposes by assigning students to create and edit encyclopaedia articles. Such assignments can have many benefits: in addition to improving students’ understanding of content, the assignments have been shown to increase their intrinsic motivation to learn, develop digital competences and build cross-cutting skills in online communication and interaction. Lastly, in connection with universities’ institutional role, editing Wikipedia can be an interesting channel for the public communication of science. more...

To gain insight into the regulatory mechanisms and the signals responsible for the adaptation of higher plants to conditions of varying sulfate availability, we have isolated from a sulfate- deprived root library maize cDNAs encoding sulfate permease (ZmST1) and ATP sulfurylase (ZmAS1), the two earliest components of the sulfur assimilation pathway. The levels of ZmST1 and ZmAS1 transcripts concomitantly increased in both roots and shoots of seedlings grown under sulfate-deprived conditions, and rapidly decreased when the external sulfate supply was restored. This coordinate response, which was not observed under conditions of limiting nitrate or phosphate, correlated with the depletion of glutathione, rather than sulfate stores. However, drastically reducing glutathione levels through treatment with buthionine sulfoximine, a specific inhibitor of γ-glutamyl cysteine synthetase, did not provide an adequate stimulus for the up- regulation of either sulfate permease or ATP sulfurylase messengers. Indeed, L-cysteine, but not D-cysteine, effectively down-regulated both transcripts when supplied to sulfur-deficient seedlings under conditions of blocked glutathione synthesis. Altogether, these data provide evidence for the coordinate regulation of sulfur assimilation mRNAs in higher plants and for the glutathione-independent involvement of cysteine as a stereospecific pretranslational modulator of the expression of sulfur status-responsive genes. more...

EDITORIAL COMMENT: This issue of the journal contains 2 case reports of interstitial ectopic pregnancies, both treated by injection of methotrexate, by intramuscular injection in one case, and by laparoscopically-guided injection into the ectopic pregnancy mass in the other. The methods were not equally successful. Parker and colleagues reviewed the published data and concluded that single-dose intramuscular methotrexate for treatment of ectopic pregnancy is associated with a high failure rate, and serious complications have occurred (11). The authors of this report advocate the use of laparoscopically-guided local methotrexate infiltration. The selection of which cases of ectopic pregnancy warrant this regimen is not yet established in the editor's opinion. Ironically one of the reviewers of this case report, selected because of his known experience with ultrasonographically-guided direct local methotrexate injection into early tubal ectopic pregnancies, stated that, because of the risk of causing haemorrhage, that he had preferred methotrexate by intramuscular injection in 2 women with interstitial ectopic pregnancies, who then, at a later date, had excision of the ectopic pregnancies performed laparoscopically. Summary: This case describes the use of ultrasound to diagnose an interstitial ectopic pregnancy and the subsequent management and ultrasound follow-up. Interstitial pregnancy is a rare but potentially life-threatening complication of pregnancy. Management options depend upon the timing of diagnosis, and the desire for maintenance of fertility. Methotrexate injection, under ultrasonographic or laparoscopic guidance, seems to be the management of choice when the ectopic is diagnosed at an early stage. more...

Background Therapeutic embolization of the uterine arteries has been successfully used to manage profuse gynecological hemorrhage. In the present study we aimed to investigate whether embolization of uterine arteries may serve as a safe and effective alternative treatment in cases of menorrhagia in fertile and perimenopausal women. As a first step, we have evaluated the methodology, patient reactions and effects on the uterine vasculature. Methods The distal part of the uterine artery was embolized with polyvinyl alcohol particles via catheterization of the right femoral artery. Total abdominal hysterectomy was performed the next day. Results Bilateral embolization in two patients resulted in considerable pain that required morphine analgesic medication and epidural analgesia. One patient was embolized unilaterally and experienced only slight discomfort with no need for analgesic medication at all, indicating that unilateral embolization is a well-tolerated method. After embolization, angiography showed stagnant flow in embolized vessels without contrast filling of distal branches. Angiography of the specimen showed normal vascular architecture in non-treated vessels. In treated vessels the main arterial trunks were patent but all smaller branches were occluded. Histology showed that most of the particles lodged in small arteries and that arterioles never showed injected material. Conclusion The study indicates that the procedure involves an efficient occlusion of uterine vessels and that unilateral embolization of uterine arteries is well tolerated. more...

Therapeutic embolization of the uterine arteries has been successfully used to manage profuse gynecological hemorrhage. In the present study we aimed to investigate whether embolization of uterine arteries may serve as a safe and effective alternative treatment in cases of menorrhagia in fertile and perimenopausal women. As a first step, we have evaluated the methodology, patient reactions and effects on the uterine vasculature.The distal part of the uterine artery was embolized with polyvinyl alcohol particles via catheterization of the right femoral artery. Total abdominal hysterectomy was performed the next day.Bilateral embolization in two patients resulted in considerable pain that required morphine analgesic medication and epidural analgesia. One patient was embolized unilaterally and experienced only slight discomfort with no need for analgesic medication at all, indicating that unilateral embolization is a well-tolerated method. After embolization, angiography showed stagnant flow in embolized vessels without contrast filling of distal branches. Angiography of the specimen showed normal vascular architecture in non-treated vessels. In treated vessels the main arterial trunks were patent but all smaller branches were occluded. Histology showed that most of the particles lodged in small arteries and that arterioles never showed injected material.The study indicates that the procedure involves an efficient occlusion of uterine vessels and that unilateral embolization of uterine arteries is well tolerated. more...

LDL-apheresis often induces an almost constant and progressive increase of the differential pressure of plasma flowing through the dextran sulphate cellulose column, reducing the efficacy of the treatment. On two occasions we were able to identify a fibrin plug by immunofluorescence. Our aim was to verify the modification of some coagulation indicators in patients undergoing LDL-apheresis and whether an activation of coagulation occurs in the LDL-apheresis device. Blood samples were obtained from six patients with familial hypercholesterolaemia who were undergoing LDL-apheresis. During the same session further blood/ plasma samples were taken from the LDL-apheresis device at different sites and at different volumes of filtered blood. In patients after LDL-apheresis the following modifications were found: a 25% decrease of fibrinogen and a slight increase in F1 + 2 plasma levels. No relevant changes in thrombin-antithrombin complexes and fibrinopeptide A plasma levels were noted. In the LDL-apheresis device the main results were: (a) fibrinogen was trapped in the dextran sulphate cellulose column in the early phases; (b) activation of coagulation was recognisable in the plasma separator during the procedure and progressively increased with duration of LDL-apheresis; (c) thrombin-antithrombin complexes, formed in the plasma separator, were retained by the dextran sulphate cellulose column. In conclusion, LDL-apheresis activates coagulation in the device. Shortening cycle time or using nafamostat mesilate as an anticoagulant, could be interesting alternatives for improving the procedure. more...

Primary infra-inguinal arterial reconstructions were reviewed for primary patency and outcome of thrombosis in 144 patients. Distal anastomoses in these patients were to the popliteal artery and were above the knee in 63, below the knee in 53 and at the tibial level in 28. The treatment used was: polytetrafluoroethylene (PTFE) in 33 cases, PTFE with an interposition vein cuff in 29 cases, autogenous saphenous vein (ASV) in situ in 47 cases, and reversed technique in 26 cases. Life table analysis showed a 59% overall primary patency at 3 years. Patency rates of above knee anastomoses (65%) and below knee (61%) were statistically different from the tibial anastomoses (42%. P = 0.005). In both above and below knee popliteal anastomoses there was a statistically significant difference in the patency of ASV and the PTFE/vein cuff technique (P= 0.0006) but there was no difference between ASV and FTFE. There was no difference in patency rates for the various types of grafts with tibial anastomoses. Data were analysed at 3 years, taking into account the variables of smoking, diabetes or indications for surgery respectively and no difference was found in patency. The number and calibre of the run-off vessels did not influence patency significantly, hence anastomosis to any good quality vessel regardless of run-off is recommended. The poor results with the interposition vein cuff technique are unexplained but this study suggests that the technique should he reserved for anastomoses below the popliteal artery. more...

The mutual interaction between monocytes and low density lipoprotein (LDL) in atherogenesis prompted a test of the hypothesis that LDL-apheresis could reduce the adhesive properties of monocytes to endothelium; and therefore interfere with a key mechanism in atheroma formation. Five patients affected by heterozygous familial hypercholesterolemia were studied. All patients received LDL-apheresis treatment with selective adsorption of LDL-cholesterol on dextran-sulphate columns. Low density lipoprotein particles were isolated by sequential preparative ultracentrifugation and subfractionated by ion exchange high performance liquid chromatography. Thiobarbituric acid reacting products of lipid peroxidation were measured fluorometrically. Vitamin E was estimated by high performance liquid chromatographic technique. Monocytes were isolated from patients blood before and 1 day after LDL-apheresis by Percoll gradient. The blood samples for monocyte adhesion were drawn from control subjects for 2 consecutive days. The adhesion of monocytes to an endothelial monolayer was evaluated by assaying the peroxidase content of the adherent monocytes. Low density lipoprotein-apheresis reduced total cholesterol (-65%; p < 0.01), LDL-cholesterol (-75%; p < 0.01), triglycerides (-51%; p < 0.05), and fibrinogen (-28%; p < 0.01). With LDL-apheresis treatment, a reduction of 54% in oxidized LDLs was observed; vitamin E concentration significantly increased in LDLs (+ 14.2%; p < 0.05). The monocyte adhesion decreased by approximately 61% after apheresis; the variation became statistically significant (-65%; p < 0.01) when endothelial cells were stimulated by lipopolysaccaride. more...

Objective: To determine the live birth rate following surgical reversal of sterilisation in women aged 40 years and older. Design: Retrospective cohort study of pregnancy outcome following day surgery microsurgical reversal of sterilisation performed by two reproductive microsurgeons in the private sector. Setting and patients: 47 patients (aged 40 years or older) who had reversal of sterilisation performed between 1997 and 2005 in Adelaide, South Australia (n = 35), or the Infertility Centre of St Louis, Missouri, USA (n =1 2). Main outcome measures: Independently audited live birth surviving the neonatal period. Results: Of the 47 patients on whom follow-up was obtainable from the two centres, 19 (40%) had a live birth, 7 had had only a first trimester miscarriage at the time of followup, and 21 (44%) had failed to conceive. Age at conception ranged between 40 and 47 years. Two women had two live births following surgery. The total direct costs (Australian dollars, adjusted to 2005) in Australia were $4850 per treatment, and $11 317 per live birth. The corresponding direct cost of a single cycle of in-vitro fertilisation (IVF) in Australia has been estimated at $6940, with a cost per live birth of $97 884 for women aged 40–42 years and $182 794 for older women. Conclusion: Previously sterilised women wanting further pregnancy should be offered tubal surgery as an alternative to IVF, as it offers them the opportunity to have an entirely natural pregnancy. In settings where IVF is financially supported by government agencies or insurance, tubal reversal is a highly cost-effective strategy for the previously more...

To the Editor. —Dr Agishi 1 discusses the evidence that anaphylactoid reactions could occur during low-density lipoprotein (LDL) apheresis performed with the dextran sulfate— cellulose (DSC) adsorption columns 2 in patients taking angiotensin converting enzyme (ACE) inhibitors. 3 This adverse reaction is triggered by the entrapment of the Hageman factor, high-molecular-weight kininogen, and prekallikrein into the DSC column, leading to kallikrein activation and bradykinin generation. Bradykinin is usually inactivated rapidly by kinases I and II, 3 which are identical to ACE and are inhibited by ACE inhibitors. Therefore, increased generation and decreased catabolism of bradykinin may explain this adverse reaction during the LDL apheresis. Recently, we observed a case of hypotension in a patient treated with an ACE inhibitor during an LDL-apheretic procedure without the DSC column (sham apheresis). After obtaining informed consent, we routinely perform sham apheresis in candidates for chronic LDL apheresis to check its feasibility (eg, adequate more...

A very fast, highly efficient, versatile and low-cost cloning of PCR products is described. PCR amplicons, obtained with any set of primers, is directly integrated into circular plasmid vectors by means of a one-step restriction-ligation procedure. When using proof-reading DNA polymerases, 100% cloning efficiency is easily achieved, implying that direct cloning into 'final-use' vectors (i.e. avoiding any intermediate cloning step) is a feasible task. Albeit with a lower efficiency, the same procedure is also suitable for the cloning of PCR products generated by 'non-proof-reading' DNA polymerases. Furthermore, with a simple modification of the vector polylinker site, the present method can be easily adapted to the directional cloning of open-reading-frame-encoding amplicons. This one-step procedure thus couples high efficiency with high reliability and versatility, and lends itself as the method of choice for routine cloning of PCR products. more...

OBJECTIVE To assess the efficacy of destruction of the endometrium by ablation with a neodymium:YAG laser or resection with electrocautery in the management of menorrhagia. DESIGN The efficacy of the treatment was assessed subjectively by each patient comparing the duration and amount of her menstrual bleeding before and after the operative procedure. Randomisation of treatments or patients was not undertaken. SETTING The study was carried out in tertiary centres. Most of the procedures were undertaken in the Day Surgery Units of those centres. PATIENTS A total of 64 patients underwent one or more procedures. One had continuous bleeding per vagina on hormone replacement therapy, all others had menorrhagia which they considered incapacitating. INTERVENTIONS Of the patients 40 had removal of endometrium via a resectoscope and 24 had endometrial vaporisation with the neodymium:YAG laser. MAIN OUTCOME MEASURE The outcome was assessed by each patient as amenorrhoea, hypomenorrhoea, attainment of normal menstrual amount or no change in menstrual loss. RESULTS Both operative procedures were satisfactory in that amenorrhoea, hypomenorrhoea or normal menstruation was achieved in 87% of cases. Uterine perforation occurred on two occasions with thermal bowel injury in one patient. CONCLUSION Destruction of the endometrium by the neodymium:YAG laser or the resectoscope is a successful way of managing menorrhagia in the absence of demonstrable pathological cause in nearly 90% of cases. The performance of the procedures as day cases with minimal discomfort makes them attractive alternatives to hysterectomy in the management of menorrhagia. more...

The identification of plant genes involved in early phases of in vitro morphogenesis can not only contribute to our understanding of the processes underlying growth regulator-controlled determination, but also provide novel markers for evaluating the outcome of in vitro regeneration experiments. To search for such genes and to monitor changes in gene expression accompanying in vitro regeneration, we have adapted the mRNA differential display technique to the comparative analysis of a model system of tomato cotyledons that can be driven selectively toward either shoot or callus formation by means of previously determined growth regulator supplementations. Hormone-independent transcriptional modulation (mainly down-regulation) has been found to be the most common event, indicating that a non-specific reprogramming of gene expression quantitatively predominates during the early phases of in vitro culture. However, cDNA fragments representative of genes that are either down-regulated or induced in a programme-specific manner could also be identified, and two of them (G35, G36) were further characterized. One of these cDNA fragments, G35, corresponds to an mRNA that is down-regulated much earlier in callus- (day 2) than in shoot-determined explants (day 6). The other, G36, identifies an mRNA that is transiently expressed in shoot-determined explants only, well before any macroscopic signs of differentiation become apparent, and thus exhibits typical features of a morphogenetic marker. more...

Summary: This study presents an independently-assessed comparison of the laparoscopic view obtained using a 2mm versus 10mm laparoscope in women with suspected pelvic pathology. Fifteen female volunteers booked for laparoscopy with clinical evidence of pelvic abnormality according to clinical findings and/or pelvic ultrasound were recruited for this study. Sequential observations were carried out by independent observers for clinically significant differences. Although discrepancies were noted in 3 patients the view obtained with the 2mm microendoscope was considered to be comparable to that obtained with the 10mm telescope. The cases with discordant findings included mild or minimal endometriosis and distal tubal disease. The results of this study suggest that microendoscopy is likely to be entirely adequate for many routine laparoscopic procedures and sterilization. more...

Blood flow to the ovary, vascular pedicle and oviduct was measured in anaesthetized non-cycling and cycling ewes by timed collection of ovarian venous blood. The degree of arterio-venous shunting across the ovary and pedicle was estimated both in vivo and in vitro by perfusing the tissues with 15 ± 5 μm radioactive microspheres. The mean ovarian blood flow in non-cycling animals was 1.9 ml/min, which was 51% of blood flow in the ovarian vein. In cycling animals ovarian blood flow at midcycle was 2.9 ml/min (66% of ovarian venous flow) in non-luteal ovaries and 4.3 ml/min (79% of venous flow) in luteal ovaries. The degree of arterio-venous shunting was low in all stages of the cycle (1.0–2.6% across ovary + pedicle). The degree of shunting was also found to be very small in vitro (0.007–1.38%) in both non-luteal and luteal ovaries. A considerable number of microspheres was entrapped in the vascular pedicle of the ovary indicating the presence of an extensive capillary bed. There was an inverse relationship between blood flow in the ovary and flow in the vascular pedicle. Alterations in distribution of blood flow between the ovary and adjacent structures supplied by the ovarian artery may be of functional significance in allowing rapid changes in ovarian blood flow. The results of the present study indicate that changes in ovarian blood flow during the oestrous cycle are not caused by an action on arteriovenous shunt vessels. more...

In order to identify the sequence of hormonal changes surrounding farrowing in sows, concentrations of sex steroids (pregnenolone sulphate, pregnenolone, progesterone, dehydroepiandrosterone, androstenedione, testosterone, oestrone sulphate, oestrone and oestradiol), relaxin and cortisol were determined in blood samples collected via chronically implanted utero-ovarian (UOV) venous cannulae in seven animals between 120 h before and 120 h after the onset of delivery. The relaxin content of UOV plasma reached maximum values 1–2 days before delivery, at which time progesterone values were falling, or had fallen, to nadir. High concentrations of pregnenolone sulphate, an important luteal precursor to progesterone synthesis in pregnant sows, were also maintained in the UOV during the phase of progesterone decline, suggesting that progesterone secretion by ovaries of preparturient sows is constrained by enzyme activity (arylsulphatase and/or 3β-hydroxysteroid dehydrogenase), rather than by substrate availability. Amounts of other steroids in UOV plasma declined after the onset of delivery. Asynchronies in flux of progestagens and of relaxin in the UOV drainage of periparturient sows portray a sequence of changes in luteal secretory activity prior to delivery. During farrowing, times of expulsion of piglets did not consistently coincide with pulses of release of any particular steroid hormone, or of relaxin. more...

Summary: Ninety-eight patients were given prostaglandin F2α for the induction of mid-trimester abortion. In 42 the PGF2α was administered by the extra-amniotic route and 37 aborted. A further 56 received the PGF2α intra-amniotically and 55 aborted. Dose regimens are compared. more...

Renal biopsies were performed on 11 patients considered clinically and histologically to have pre-eclampsia. Immunofluorescent studies with fluorescein-labelled anti-IgG, IgA, IgM, and IgE complement, albumin, fibrin, and fibrinogen were carried out on the tissue obtained. Significant correlation was obtained between the clinical severity of the disease and the density and pattern of IgM and IgG deposition. Complement was found in glomeruli in severe cases, while complement deposition in the walls of afferent and efferent arterioles was a constant finding. These findings support the concept that an immunological mechanism may be responsible for the renal lesions in pre-eclampsia. If immunity does play a part in the pathogenesis of pre-eclampsia, possible mechanisms include the involvement of histocompatibility antigens and cross-reactivity of fetal and maternal tissues. Renal fibrin deposition in pre-eclampsia may be secondary to an immune process, an occurrence well-described in other forms of glomerulonephritis in man and other species. more...

This study investigated the inhibition by colostrum of the uptake of tritiated thymidine by both autologous and heterologous peripheral blood mononuclear cells (PBMC) stimulated with the mitogens phytohaemagglutinin (PHA), pokeweed mitogen (PWM) or Concanavalin A (Con A). This inhibition was not the result of an alteration in the dose response or kinetics of mitogenesis by the colostrum. The inhibitory factor in the colostrum was neither dialysable nor lymphocytotoxic. Lymphocytes which were briefly pre-treated with colostrum, then washed and cultured, also incorporated much less labelled thymidine than control cells. The inhibitory factor was not adsorbed from colostrum by mononuclear cells. We conclude that colostrum contains a cytostatic factor which inhibits mitogenesis in PBMC. more...