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Abstract: Ethanolamine ammonia-lyase (EAL) is an adenosylcobalamin (AdoCbl, a B12 coenzyme) dependent enzyme expressed in enteric bacteria, which catalyzes conversion of ethanolamine in ammonia and acetaldehyde under anaerobic conditions. The most accepted reaction mechanism includes five steps: (a) 5-deoxyadenosyl radical (Ado ) generation by homolytic cleavage of a Co–C bond in AdoCbl; (b) substrate activation by Ado H-abstraction from C1; (c) 1,2-NH2 migration in substrate’s radical; (d) Ado regeneration; (e) NH3 elimination from substrate leaving acetaldehyde as final product. Steps a–c have been the subject of both empirical studies and computational modeling performed at the ab initio and DFT levels, with different representations of the protein environment, attempting to explain the observed kinetics by total/partial protonation or by “push–pull” catalysis assisted by two amino acid residues. In such models, a drastic reduction of the reactive system and of the influence of the enzyme environment was introduced, the latter mainly due to the lack of detailed data on the 3D structure of EAL (note elucidated until 2007) and the nature of the active site (for which a structure of the substrate-EAL complex in presence of adeninylpentylcobalamin has been very recently reported, shedding light on its structural features). Here we assess at the PCM//UB3LYP/6-31G(d,p) level how using a complete molecular description of both reactants in step b and introducing the polarizing effect of the protein/cofactor environment (mimicked through a continuum model with ε =10) impacts on the nature and energetics of the intermediate complexes (ICs) and transition states (TSs) under three catalytic scenarios (total, partial, and no protonation). Our results show that a concerted asynchronous mechanism involving distonic radical species is at play, passing through quasi-linear early TSs displaying a C1–H cleavage slightly more advanced than formation of the new H–C bond in Ado. Whereas the enzyme’s polarizing environment differentially stabilizes the TS lowering the barrier by ∼2.4–2.8kcal/mol, a reduction of 1.4kcal/mol in is the outcome of substrate’s partial/total protonation, without a clear distinction among them, in opposition to what previously shown based upon more simplified models. [Copyright &y& Elsevier]

There is increasing evidence that farmers in many areas are achieving below maximum yields due to insufficient pollination. Practical and effective approaches are needed to maintain wild pollinator populations within agroecosystems so they can deliver critical pollination services that underpin crop production. We established nesting and wildflower habitat interventions in 24 UK apple orchards and measured effects on flower-visiting insects and the pollination they provide, exploring how this was affected by landscape context. We quantified the extent of pollination deficits and assessed whether the management of wild pollinators can reduce deficits and deliver improved outcomes for growers over 3 years. Wildflower interventions increased solitary bee numbers visiting apple flowers by over 20%, but there was no effect of nesting interventions. Other pollinator groups were influenced by both local and landscape-scale factors, with bumblebees and hoverflies responding to the relative proportion of semi-natural habitat at larger spatial scales (1000 m), while honeybees and other flies responded at 500 m or less. By improving fruit number and quality, pollinators contributed more than £16 k per hectare. However, deficits (where maximum potential was not being reached due to a lack of pollination) were recorded and the extent of these varied across orchards, and from year to year, with a 22% deficit in output in the worst (equivalent to ~£14 k/ha) compared to less than 3% (equivalent to ~£2 k/ha) in the best year. Although no direct effect of our habitat interventions on deficits in gross output was observed, initial fruit set and seed set deficits were reduced by abundant bumblebees, and orchards with a greater abundance of solitary bees saw lower deficits in fruit size. The abundance of pollinators in apple orchards is influenced by different local and landscape factors that interact and vary between years. Consequently, pollination, and the extent of economic output deficits, also vary between orchards and years. We highlight how approaches, including establishing wildflower areas and optimizing the ratio of cropped and non-cropped habitats can increase the abundance of key apple pollinators and improve outcomes for growers., (© 2022 The Authors. Ecological Applications published by Wiley Periodicals LLC on behalf of The Ecological Society of America.)

Background In prenatal diagnosis, a thickened nuchal translucency (NT) is one of the most sensitive and specific markers for several defects but it may also be found in 5% of healthy fetuses. The pathophysiological causes that lead to an increase in NT are not yet fully understood. Metabolomics represents a new promising approach, useful for studying different metabolites in biological organisms in response to environmental stressors. The aim of our study was to investigate the metabolomic profile of the amniotic fluid samples (AFS) of euploid fetuses with enlarged nuchal translucency (ENT) compared to a control group (C group). Methods This study was carried out on a group of women who underwent second-trimester amniocentesis for advanced maternal age (C group) or for NT ≥95th percentile (ENT group) found during first-trimester aneuploidy screening. AFS were analyzed with proton nuclear magnetic resonance (1H-NMR) and high-performance liquid chromatography (HPLC), and subsequent multivariate and univariate statistical analyses were conducted, followed by pathway analysis. Results In total, 67 AFS from the C group and 23 from the ENT group were analyzed. Partial least square discriminate analysis was carried out (R2X=0.784, R2Y=0.658, Q2=0.622, P<0.0001). A different metabolic profile was observed in the ENT group compared with the C group, suggesting an energetic shift to a glycolytic phenotype in an oxidative environment in the ENT group compared to the C group. Conclusion Metabolomic studies enable the identification of metabolic alterations occurring in fetuses with ENT. These findings may provide a new basis for better understanding the pathophysiological mechanisms in this prenatal phenomenon.

Objective: To evaluate the best management of fetal megacystis diagnosed in the first trimester and define the prognosis and the most appropriate follow-up as early as possible., Methods: This is a retrospective study of first-trimester fetal megacystis diagnosed in pregnant women who performed a combined screening for fetal aneuploidy. Megacystis was defined as a longitudinal bladder diameter (LBD) greater than 7 mm. All fetuses were divided into two groups according to the LBD: Group A with LBD > 15 mm and Group B with LBD < 15 mm. The fetal karyotype and associated anomalies were evaluated. Ultrasound monitoring was performed every 2 weeks (a second ultrasound scan after 2 weeks from diagnosis and a third ultrasound scan 2 weeks after the second one)., Results: Twenty-six cases were identified between 2011 and 2016; three cases of aneuploidy were excluded from the study. Of the remaining 23 cases, 11 were included in Group A and 12 in Group B. All Group A fetuses had an adverse outcome. In Group B: five (41.7%) cases had an adverse outcome and seven (58.3%) had a spontaneous resolution of megacystis. The ultrasound findings of both the ultrasound scans, the second and the third, were 100% concordant., Conclusion: An ultrasound scan performed 2 weeks after the megacystis diagnosis can predict the outcome in fetuses with an LBD < 15 mm as early as the end of the first trimester. The outcome of euploid fetuses with an LBD < 15 mm was favorable in 58.3% of the cases.

Background: Gonadotropins are protein hormones which are central to the complex endocrine system that regulates normal growth, sexual development, and reproductive function. There is still a lively debate on which type of gonadotropin medication should be used, either human menopausal gonadotropin or recombinant follicle-stimulating hormone. The objective of the study was to perform a systematic review of the recent literature to compare recombinant follicle-stimulating hormone to human menopausal gonadotropin with the aim to assess any differences in terms of efficacy and to provide a cost evaluation based on findings of this systematic review., Methods: The review was conducted selecting prospective, randomized, controlled trials comparing the two gonadotropin medications from a literature search of several databases. The outcome measure used to evaluate efficacy was the number of oocytes retrieved per cycle. In addition, a cost evaluation was performed based on retrieved efficacy data., Results: The number of oocytes retrieved appeared to be higher for human menopausal gonadotropin in only 2 studies while 10 out of 13 studies showed a higher mean number of oocytes retrieved per cycle for recombinant follicle-stimulating hormone. The results of the cost evaluation provided a similar cost per oocyte for both hormones., Conclusions: Recombinant follicle-stimulating hormone treatment resulted in a higher oocytes yield per cycle than human menopausal gonadotropin at similar cost per oocyte.

The severity, morbidity and financial costs of atopic eczema (AE) were assessed during a 1-year prospective study of a cohort of 85 people aged 14-63 years (mean 36 years) with the disease. A dermatologist examined each participant using the Six Area Six Sign Atopic Dermatitis severity scoring system to classify severity. Participants completed a Dermatology Life Quality Index (DLQI), a Nottingham Eczema Severity Score (NESS) and an ongoing diary of health-care consultations and treatment costs. Follow up by mail to each participant was conducted every 2 months and participants completed a NESS, a DLQI and a diary of costs incurred. The DLQI data revealed that 36% spent over 10 min per day applying treatments, 28% indicated that AE influenced the clothes they wore, 21% felt embarrassed by their skin and 15% reported problems with treatments. There appeared to be a relationship between increased morbidity and increased severity. The average annual out-of-pocket cost for products used for treatment was A$425, ranging from A$13.50 to over A$2000 per individual. The average out-of-pocket cost for medical consultations was A$120, ranging from zero to over A$800 per individual. Although there were concerns about the reproducibility of the severity and morbidity measures, the data showed that AE can have substantial effects both financially and from a personal perspective for those affected.

This study assessed whether the function of an individual's problem behavior was related to the effectiveness of an intervention involving choice among tasks. Analogue functional analyses were conducted with 7 students with various diagnoses to determine whether problem behaviors were maintained by escape or attention. Following identification of the function of each student's problem behavior, reversal designs were used to assess the effectiveness of an intervention that allowed the students to choose their own instructional tasks. Results showed that students who displayed escape-maintained problem behavior showed substantial reductions in such behavior when they were provided with opportunities to choose among tasks. On the other hand, students who displayed attention-maintained problem behavior did not show any effects as a result of the choice intervention. These findings are discussed in terms of the effective use of behavior management programs involving choice and the reduction of problem behavior.

Eighty-three participants with psoriasis were followed over a 2-year period assessing the severity, morbidity and cost of their disease over time. At recruitment, they were examined by a dermatologist who classified them on a global basis as mild (47%), moderate (35%) or severe (18%). A Psoriasis Area and Severity Index (PASI) score was also recorded at the initial interview. Participants completed questionnaires on the morbidity related to having psoriasis using the Psoriasis Disability Index (PDI) and a self-administered PASI (SAPASI) score at the initial interview and at 3-monthly intervals over the 2-year period. During the 3-monthly follow ups, patients also collected information on the cost of treatment. Using the PDI data, two-thirds of the respondents said that, as a result of their psoriasis, they altered the way they carried out their normal daily activities; more than 50% wore different types or colours of clothing; more than 50% said their home was made more messy or untidy; and over a third had problems at the hairdresser or difficulties playing sport. The annual out-of-pocket expense for medical products was around AUD$250 per person, with costs ranging from zero to more than AUD$2,000 per individual over the 2-year period. Costs were highest for over-the-counter products purchased without a medical prescription. There were similar variations in the out-of-pocket expenses of medical consultations depending on the level of medical care required. The study revealed that the standard methods used for classification of severity of psoriasis, such as the PASI or SAPASI scores, do not take into account the treatment being used at the time the score is recorded and therefore may not accurately reflect the true severity.

In 1989 the Australian Council on Healthcare Standards (ACHS) embarked on a programme to develop acute health care clinical indicators in conjunction with the Australian medical colleges. Through a carefully structured stepwise process this collaboration established a 'World first' in 1993 with the introduction of the first set of indicators into the ACHS Accreditation programme. The programme remains unique in the formal involvement of providers in the development process and in the scope of the clinical areas covered in acute health care. From the year 2000 there will be 18 sets (and over 200 indicators) from which health care organisations (HCOs) can choose to monitor the major services they provide. There remains no compulsion to address a specific number of indicators. The growth of the programme has been considerable with more than half of the nations' acute HCOs reporting their clinical indicator data (twice yearly) and it provides a reflection of the care given for the majority of patient separations in acute care. This reporting process allows HCOs to receive feedback on the aggregate results together with comparative peer group information for each indicator they address. In addition to numerous publications in peer reviewed journals an annual aggregate report, 'the Measurement of Care in Australian Hospitals' is published. It reports both qualitative and quantitative data on all indicator sets for the preceding year. Validity of the indicators is strengthened each year with a review process and reliability and reproducibility of the data can now be demonstrated. The clinical response to the indicators has been overwhelming and there is now documented evidence of numerous actions taken by HCOs to improve both the processes and the outcomes of patient care. The nation wide database can be expected to reflect trends in care over the next few years. The process of indicator refinement, however, will continue and it is likely that a reduction in the total number of indicators will occur with a core group of the more 'robust' indicators remaining. Further directions in indicator development are likely to be in the area of multidisciplinary care and in the assessment of longer-term outcomes. In addition to measures of the quality of care, hopefully, in time, health care providers will also take part in the establishment of measures of the appropriateness of that care.

The high sensitivity of pentagastrin stimulation in detecting primary or metastatic medullary thyroid cancer (MTC) suggests widespread expression of the corresponding receptor type on human MTC. Indeed, autoradiographic studies demonstrated cholecystokinin (CCK)-B/gastrin receptors not only in >90% of MTCs but in a high percentage of small cell lung cancers and potentially a variety of gastrointestinal adenocarcinomas. In a pilot study, we have demonstrated the feasibility of radiolabeled gastrin-I to target CCK-B receptor-expressing tissues in vivo in animals and patients (T. M. Behr et al., Eur. J. Nucl. Med., 25: 424-430, 1998). The aim of the present study was to systematically optimize, in a preclinical model, suitable radioligands for targeting CCK-B receptors in vivo. For this purpose, a variety of CCK/gastrin-related peptides, all having in common the COOH-terminal CCK-receptor binding tetrapeptide sequence Trp-Met-Asp-PheNH2 or derivatives thereof, were studied. They were radioiodinated by the Iodogen or Bolton-Hunter procedures. The peptides tested were members of the gastrin- or cholecystokinin families or possessed characteristics of both, which differ by the intramolecular position of a tyrosyl moiety (occurring in native or sulfated form). Their stability and affinity were studied in vitro and in vivo; their biodistribution and therapeutic efficacy were tested in nude mice bearing s.c. human MTC xenografts. Diethylene-triamine-pentaacetate derivatives of suitable peptides were synthesized, evaluated, and labeled with (111)In. All members of the CCK or gastrin family were stable in serum (with t(1/2)s of several hours at 37 degrees C); nevertheless, the stability of those peptides was highest that bore the NH2-terminal pGlu residues (e.g., big gastrin, gastrin-I, caerulein, and others) or D-amino acids. In accordance to their comparably low affinity, nonsulfated members of the CCK family showed fairly low uptake in the tumor and other CCK-B receptor-expressing tissues (e.g., the stomach). Sulfated CCK derivatives performed significantly better but additionally displayed a high uptake in normal, CCK-A receptor-expressing tissues (such as the liver/gallbladder, pancreas, and bowel). Best tumor uptake and tumor:nontumor ratios were obtained with members of the gastrin family, probably because of their selectivity and affinity for the CCK-B receptor subtype. Pilot therapy experiments in MTC bearing animals showed significant antitumor efficacy as compared with untreated controls. (111)In-Labeled diethylene-triamine-pentaacetate derivatives of minigastrin showed excellent targeting of CCK-B receptor-expressing tissues in animals and a normal human volunteer. These data suggest that CCK/gastrin analogues may be a useful new class of receptor binding peptides for diagnosis and therapy of CCK-B receptor-expressing tumors, such as MTC or small cell lung cancer. Nonsulfated gastrin derivatives may be preferable because of their CCK-B receptor selectivity, and hence, lower accretion in normal CCK-A receptor-expressing organs. Further preclinical as well as clinical studies are ongoing.

Unlabelled: The high sensitivity of pentagastrin stimulation in detecting primary or metastatic medullary thyroid cancer (MTC) suggests widespread expression of the corresponding receptor type on human MTC. Indeed, autoradiographic studies have demonstrated cholecystokinin (CCK)-B/gastrin receptors not only in more than 90% of MTCs but also in a high percentage of small cell lung cancers, some ovarian cancers, astrocytomas and potentially a variety of adenocarcinomas. The aim of this study was to systematically screen and optimize, in a preclinical model and a pilot clinical study, suitable radioligands for targeting CCK-B receptors in vivo., Methods: A variety of CCK/gastrin-related peptides, all bearing the C-terminal CCK receptor-binding tetrapeptide sequence Trp-Met-Asp-PheNH2 or derivatives thereof, were studied. They were radioiodinated by the lodogen or Bolton-Hunter procedures. The peptides were members of the gastrin or CCK families, which differ by the intramolecular position of a tyrosyl moiety. Their stability and affinity were studied in vitro and in vivo; their biodistribution and therapeutic efficacy were tested in nude mice bearing subcutaneous human MTC xenografts. Diethylenetriamine pentaacetic acid (DTPA) derivatives of suitable peptides were synthesized successfully, and their preclinical and initial clinical evaluations were performed, labeled with 111In., Results: All members of the CCK or gastrin families were stable in serum (with half-lives of several hours at 37 degrees C); nevertheless, the stability of those peptides bearing N-terminal pGlu residues or D-amino acids was significantly higher. In accordance with their comparably low affinity, nonsulfated members of the CCK family showed fairly low uptake in the tumor and other CCK-B receptor-expressing tissues. Sulfated CCK derivatives performed significantly better but also displayed a comparably high uptake in normal CCK-A receptor-expressing tissues. This effect was probably due to their similar affinity for both CCK-A and CCK-B receptors. Best tumor uptake and tumor-to-nontumor ratios were obtained with members of the gastrin family because of their selectivity and affinity for the CCK-B receptor subtype. Pilot therapy experiments in MTC-bearing animals showed significant antitumor efficacy compared with untreated controls. DTPA derivatives of minigastrin were successfully developed. In a pilot clinical study, radioiodinated and 111In-labeled derivatives showed excellent targeting of physiological CCK-B receptor-expressing organs, as well as all known tumor sites., Conclusion: CCK/gastrin analogs may be a useful new class of receptor-binding peptides for diagnosis and therapy of CCK-B receptor-expressing tumors, such as MTC or small cell lung cancer. Nonsulfated gastrin derivatives may be preferable because of their CCK-B receptor selectivity, hence lower accretion in normal CCK-A receptor-expressing organs.

The outstanding sensitivity of pentagastrin in detecting the presence of primary, recurrent or metastatic medullary thyroid cancer (MTC) suggests widespread expression of the corresponding receptor type in human MTC. Indeed, recent autoradiographic studies have demonstrated the presence of cholecystokinin (CCK)-B (= gastrin) receptors not only in more than 90% of MTCs but also in a high percentage of small cell lung cancers, stromal ovarian cancers, astrocytomas and several other tumour types. The aim of this study was to evaluate whether radiolabelled gastrin may be suitable for targeting CCK-B receptor-expressing tumours in vivo. For this purpose, the biodistribution of the radioiodinated human heptadecapeptide gastrin-I was studied in nude mice bearing subcutaneous xenografts of the human MTC cell line, TT. Initial therapy experiments were undertaken. Finally, the biodistribution of iodine-131- labelled gastrin-I was studied in a patient with metastatic MTC. At a peptide amount of approximately 1 microg, maximum tumour uptake (8.9+/-2.9%ID/g) was observed in animals at 1 h post injection, with tumour-to-blood ratios as high as 6.3+/-1.9. Physiological CCK-B receptors in the stomach, gallbladder and pancreas of the mice were targeted as well. The major route of excretion was renal, but strong evidence for a biliary excretion pathway also exists. Pilot therapy studies with 131I-labelled gastrin showed significant anti-tumour efficacy as compared with untreated controls. In accordance with the preclinical data, good receptor targeting was observed in the tumour sites, stomach, gallbladder and pancreas of a patient with metastatic MTC. These data suggest that gastrin and its analogues may represent a useful new class of receptor binding peptides for diagnosis and therapy of a variety of tumour types, including MTC and small cell lung cancer. Future preclinical and clinical studies will address in more detail the molecular features that render CCK-B receptor binding agents potentially useful candidates for in vivo scintigraphy and radionuclide therapy.

The intravenous therapy team at Massachusetts General Hospital studied the potential infectious risks of maintaining percutaneous inserted central catheters (PICC) for prolonged periods. Cultures of 100 PICC sites and catheters were performed on removal of the catheters, which had remained in place for 2 to 43 days. The insertion sites and/or proximal or distal segments of the catheters were found to be colonized in 11% of the patients, with distal catheter tips significantly colonized in only four patients. Nine of the patients were colonized at the insertion site. Bacteremia did not occur as a result of the extended dwell time of the catheters.

English sole (Parophyrs vetulus) in Puget Sound, Washington, USA are at risk of hepatocarcinogenesis specifically in areas adjacent to polluting industrial effluents. A question concerning population and ecosystem survival is whether any of the effects of etiopathologic change are reversible. This has been approached by looking for evidence of tumor accelerating effects in an exposed population. Cellular parameters were determined by flow cytometry for hepatocytes of English sole. Cells containing hyperdiploid DNA not present in fish from reference waters, Port Madison, were found in all non-tumor-bearing and tumor bearing fish taken from a polluted site, Eagle Harbor, where incidence of hepatic neoplasia approaches 30%. Induction of altered DNA content in the exposed general hepatocyte population suggests environmental induction rather than an association with lesions per se. In contrast, glutathione levels in hepatocytes (0.8-3.2 nmol/mg protein), were little influenced by the exposure site, consistent with the apparent lack of protection against chemically induced carcinogenesis in English sole. Association of altered DNA content with exposure site is significant for its potential contribution to biological acceleration and evidence of tumor promotion found at the tissue and organismic levels. The results support the notion that hepatocarcinogenesis in English sole in Eagle Harbor has a multi-year exposure etiology, in which potentially reversible accelerating influences have a role, and that glutathione conjugation is an inadequate mode of detoxification for these fish.

1. Rainbow trout (Salmo gairdneri) beta 1----4galactosyltransferase has been characterized and its properties compared to the mammalian enzyme. 2. Transfer of galactose to lactotriaosylceramide (Lc3) was optimal in the presence of 0.12% Triton CF-54 and Mn2+, in a pH range of 6.5-7.5. 3. Apparent Km values for donor UDPgalactose and acceptor Lc3 were determined to be 40 and 38 microM, respectively. 4. Glycolipid substrate specificity was found with only Lc3 being an efficient acceptor. Transfer of galactose to glycosylceramide, efficient with mammalian enzyme, was 42-fold slower. 5. In contrast to mammalian enzyme, rainbow trout beta 1----4galactosyltransferase was found to be independent from regulation by alpha-lactalbumin by the inability of alpha-lactalbumin-Sepharose to bind the enzyme and by failure of alpha-lactalbumin to inhibit the enzyme activity.