Mechanism underlying the modulation of milk production by incomplete milking.

Mammary gland secretory activity is modulated by systemic and local factors; however, the relationship between these factors is unknown. The aim of this study was to determine how a local factor, such as incomplete milking, affects mammary epithelial cell activity, number, and responsiveness to blood prolactin (PRL). Eight cows in mid-lactation were differentially milked (i.e., their right quarters were milked incompletely at approximately 70%, and their left quarters were milked completely, twice daily for 4 wk). Throughout the experiment, milk yield was measured at the quarter level. Milk samples were collected from each quarter once a week to assess the milk components, and epithelial cell concentrations, as well as to isolate milk fat globule RNA. In the weeks before and after the experiment, mammary gland functional capacity was evaluated by measuring the volume of milk harvested after complete filling of the gland. At the end of the last experimental week, mammary gland biopsies were performed on each rear quarter. The milk production of quarters milked completely remained stable during the treatment period, whereas, as expected, the milk production of quarters milked incompletely was only 53% of completely milked quarters at the end of the period. Accordingly, the expression of genes related to milk synthesis (CSN2, LALBA, and ACACA) in milk fat was lower in the quarters that were milked incompletely. Incomplete milking decreased the milk lactose content, indicating a loss of integrity of tight junctions. The total yield of epithelial cells in milk was not affected, but their concentration in milk, the BAX:BCL2 gene expression ratio, and the loss of mammary functional capacity were greater in the quarters milked incompletely, suggesting an acceleration of involution in those quarters. The expression of the short isoform of the PRL receptor gene (PRLR) tended to be lower, and the expression of STAT5A and STAT5B tended to decline in the quarters milked incompletely. In mammary gland biopsy samples, the number of both short and long isoforms of the PRLR were not affected, nor were the amount and activation of STAT3 and STAT5. However, the ratio of PRLR short isoform to PRLR long isoform was lower in the quarters milked incompletely. The decrease in milk yield induced by incomplete milking is rapid and associated with a decrease in mammary epithelial cell activity and a decrease in the number of secretory epithelial cells. The results of this experiment provide only limited support for the hypothesis that modulation of the mammary gland's responsiveness to PRL is part of the mechanism by which local factors, such as incomplete milking, modulate milk synthesis.

Is cestode infection intensity associated with decreased body condition in the Eurasian woodcock Scolopax rusticola?

The Eurasian woodcock Scolopax rusticola is a widespread woodland specialist and a widely harvested quarry species throughout its European wintering areas, including Britain. Woodcock are prone to cestodiasis, but prevalence levels and possible effects on body condition remain under-studied. We studied the prevalence, abundance and intensity of cestodiasis in 161 woodcock harvested in four British regions in December and January during two consecutive winters (2013/14 and 2014/15). Cestodiasis prevalence was 90%, and there was no difference in prevalence between birds harvested in Cornwall, Wessex, East Anglia and Scotland. High prevalence levels were explained by the fact that earthworms (Lumbricidae) are intermediate hosts for some cestode species and also the most important dietary component of woodcock. The distribution of cestodiasis in woodcock was aggregated, such that when using the total length of cestodes per sample to measure abundance, 65% of the birds had less than 80 cm. Cestodiasis abundance varied between sexes across regions but the intensity was not affected by region, sex, age or their interactions. The intensity of cestodiasis was positively correlated with fresh weight and pectoral mass, while no significant correlation was found with the abdominal fat pad. Our results suggest that, despite high prevalence levels and intensity of cestodiasis in woodcock, host body condition is not significantly affected and hence it is unlikely that cestodiasis has a major effect on woodcock population dynamics.

A strategy for monitoring and evaluating massive open online courses.

We argue that the complex, innovative and adaptive nature of Massive Open Online Course (MOOC) initiatives poses particular challenges to monitoring and evaluation, in that any evaluation strategy will need to follow a systems approach. This article aims to guide organizations implementing MOOCs through a series of steps to assist them in developing a strategy to monitor, improve, and judge the merit of their initiatives. We describe how we operationalise our strategy by first defining the different layers of interacting agents in a given MOOC system. We then tailor our approach to these different layers. Specifically, a two-pronged approach was developed, where we suggest that individual projects be assessed through performance monitoring; assessment criteria for which would be defined at the outset to include coverage, participation, quality and student achievement. In contrast, the success of an overall initiative should be considered within a more adaptive, emergent evaluation inquiry framework. We present the inquiry framework we developed for MOOC initiatives, and show how this framework might be used to develop evaluation questions and an assessment methodology. We also define the more fixed indicators and measures for project performance monitoring. Our strategy is described as it was developed to inform the evaluation of a MOOC initiative at the University of Cape Town (UCT), South Africa.

CUSUM method for construction of trainee spinal ultrasound learning curves following standardised teaching.

Spinal ultrasonography is a promising aid for epidural insertion. We aimed to determine the learning curve of spinal ultrasonography tasks and the number of training scans required to reach competency after undergoing standardised step-wise teaching. Trainees were required to complete a minimum of 60 assessed scans on selected non-pregnant models following attendance at two training sessions, with feedback from an expert after each scan. Learning curves were plotted using the non-risk cumulative summation technique and an acceptable failure rate of 20%. Five trainees completed between 65 and 75 scans each. All trainees were competent at identifying a randomly assigned intervertebral space after a median of five scans (range one to nine) and at measuring the depth from skin to the posterior complex after a median of 10 scans (range 1 to 42). Two trainees were competent at marking an ideal needle insertion point after 55 scans, while three trainees did not attain competency. All trainees were competent after 60 scans if the tolerance was changed from five to eight millimetre for marking the needle insertion point. The average time taken to complete a scan was 163 seconds. Our study showed that after a standardised educational intervention, anaesthetic trainees are able to identify a lumbar interlaminar space easily and can measure the depth to the posterior complex after a reasonable number of additional practice scans, but experienced difficulty accurately marking the needle insertion point whilst using spinal ultrasonography. We confirmed that it was hard to achieve competency in all aspects of spinal ultrasonography, based on assessment using our predefined competency criteria.

Shape coexistence in the neutron-deficient even-even (182-188)Hg isotopes studied via coulomb excitation.

Coulomb-excitation experiments to study electromagnetic properties of radioactive even-even Hg isotopes were performed with 2.85 MeV/nucleon mercury beams from REX-ISOLDE. Magnitudes and relative signs of the reduced E2 matrix elements that couple the ground state and low-lying excited states in Hg182-188 were extracted. Information on the deformation of the ground and the first excited 0+ states was deduced using the quadrupole sum rules approach. Results show that the ground state is slightly deformed and of oblate nature, while a larger deformation for the excited 0+ state was noted in Hg182,184. The results are compared to beyond mean field and interacting-boson based models and interpreted within a two-state mixing model. Partial agreement with the model calculations was obtained. The presence of two different structures in the light even-mass mercury isotopes that coexist at low excitation energy is firmly established.

Steatitis in wild sharptooth catfish, Clarias gariepinus (Burchell), in the Olifants and lower Letaba Rivers in the Kruger National Park, South Africa.

Large numbers of adult Nile crocodiles, Crocodylus niloticus (Laurenti), died from pansteatitis during autumn and winter 2008 in the lower Letaba and Olifants River gorge in the Kruger National Park, South Africa. Consequently, the health status of fish from these waters was investigated. The study presents the pathological findings in fish inhabiting these rivers within the boundaries of the Park. Changes typical of steatitis were diagnosed in many of the larger specimens of sharptooth catfish, Clarias gariepinus (Burchell), caught within the Olifants River gorge. These fish carried large amounts of mesenteric fat with characteristic small brown granulomata within the adipose tissue. Necrosis and inflammation of the adipose tissues, with characteristic ceroid accumulation within the resultant granulomata and the associated aggregation of ceroid-containing macrophages, were demonstrated histologically and were typical of steatitis. Other changes included mild thickening and pallor of the gill tissues and swollen, orange, fatty livers. Focal hepatic lipidosis was demonstrated histologically, and special stains revealed storage of large amounts of iron in the livers. Blood smears revealed chromatin clumping in erythrocyte nuclei and nuclear and cell membrane irregularities. This is the first record of steatitis in wild-caught C. gariepinus.

Acute intermittent porphyria and chronic renal failure.

Despite the little known association between renal damage and the acute porphyrias, limited information is available on the characteristics and pathogenesis of renal disease in this patient group. Previous reports have focused on hypertension as the principal etiological factor. We have studied a series of 9 patients with acute intermittent porphyria (AIP) attending the Porphyria Clinic at King's College Hospital, London, UK, who were referred to the Renal Unit for investigation and treatment of their renal disease. No evidence of a glomerular lesion was found in any of the patients. In contrast, renal histology showed features of a tubulointerstitial disease, and there was evidence of impaired erythropoietin production. Hypertension and nonsteroidal antiinflammatory drug use were present in about a half of the patients. It is postulated that the nephrotoxic effects of porphyrin precursors may contribute to the etiology of this clinical syndrome.

The impact of regional anaesthesia on outcome: a patient's perspective.

Peripheral nerve catheters are being used increasingly to manage acute pain. Whilst acknowledged as effective, their broader implications for patient outcome are less clear. In this case report, we describe the way in which not only was the pain management of an ischaemic leg successful via a sciatic nerve catheter, but decision-making around this strategy affected outcome.

Septicemia due to Yersinia pseudotuberculosis--a case report.

A case report is presented concerning Yersinia pseudotuberculosis septicemia presenting as an acute abdominal emergency in an elderly diabetic man with multiple medical problems.

International air travel: a risk factor for attacks in acute intermittent porphyria.

Five patients are reported with acute intermittent porphyria in whom attacks were apparently precipitated by international air travel. In four subjects this was the initial presenting attack and in a fifth the cause of an acute relapse in a patient requiring regular haem arginate prophylaxis. Multifactorial precipitants implicated include, dehydration, missed meals, alcohol use, infection, chronic hypoxia, premenstrual syndrome and stress. Acute intermittent porphyria should be suspected in individuals presenting with unexplained acute abdominal pain following international air travel. Appropriate precautions may reduce the incidence of attacks in known porphyrics.

ACP Best Practice No 165: front line tests for the investigation of suspected porphyria.

The porphyrias are uncommon disorders of haem biosynthesis and their effective management requires prompt and accurate diagnosis. This article describes methods for the determination of urinary porphobilinogen, urinary and faecal total porphyrins, and total porphyrins in erythrocytes and plasma that are suitable for use in non-specialist laboratories. The selection and interpretation of these methods, and the indications for further, more specialised, investigation are discussed.

Evidence that NADP+ is the physiological cofactor of ADP-L-glycero-D-mannoheptose 6-epimerase.

ADP-L-glycero-D-mannoheptose 6-epimerase is required for lipopolysaccharide inner core biosynthesis in several genera of Gram-negative bacteria. The enzyme contains both fingerprint sequences Gly-X-Gly-X-X-Gly and Gly-X-X-Gly-X-X-Gly near its N terminus, which is indicative of an ADP binding fold. Previous studies of this ADP-l-glycero-D-mannoheptose 6-epimerase (ADP-hep 6-epimerase) were consistent with an NAD(+) cofactor. However, the crystal structure of this ADP-hep 6-epimerase showed bound NADP (Deacon, A. M., Ni, Y. S., Coleman, W. G., Jr., and Ealick, S. E. (2000) Structure 5, 453-462). In present studies, apo-ADP-hep 6-epimerase was reconstituted with NAD(+), NADP(+), and FAD. In this report we provide data that shows NAD(+) and NADP(+) both restored enzymatic activity, but FAD could not. Furthermore, ADP-hep 6-epimerase exhibited a preference for binding of NADP(+) over NAD(+). The K(d) value for NADP(+) was 26 microm whereas that for NAD(+) was 45 microm. Ultraviolet circular dichroism spectra showed that apo-ADP-hep 6-epimerase reconstituted with NADP(+) had more secondary structure than apo-ADP-hep 6-epimerase reconstituted with NAD(+). Perchloric acid extracts of the purified enzyme were assayed with NAD(+)-specific alcohol dehydrogenase and NADP(+)-specific isocitric dehydrogenase. A sample of the same perchloric acid extract was analyzed in chromatographic studies, which demonstrated that ADP-hep 6-epimerase binds NADP(+) in vivo. A structural comparison of ADP-hep 6-epimerase with UDP-galactose 4-epimerase, which utilizes an NAD(+) cofactor, has identified the regions of ADP-hep 6-epimerase, which defines its specificity for NADP(+).

Hospital care by hospital-based and clinic-based faculty: a prospective, controlled trial.

BACKGROUND: The hospital length of stay decreases and clinical outcomes are maintained when teaching hospitals involve hospital-based attending physicians in comparison with traditional attending physicians. The attending physician's time commitment, including the number of hours per day and months per year, required to achieve this result is unknown. This study compared the clinical outcomes and cost of care for patients treated by hospital-based and clinic-based attending physicians devoting dramatically different amounts of time to supervising residents on the medical wards of a suburban county hospital. METHODS: Patients were alternately admitted to 2 groups of ward teams. Faculty who attended 10 months of the year supervised one group. The comparison group's attending physicians were on service for 2 months or less and maintained clinic responsibilities while on service. The cost of patient care was compared by means of the length of stay, total hospital costs, and costs for ancillary services. Hospital mortality and readmission rates compared clinical outcomes. RESULTS: There were 4456 patients hospitalized on the medical wards of a teaching service. No differences were detected in the length of stay (4.37 +/- 0.1 days for hospital-based and 4.39 +/- 0.1 days for clinic-based attending physicians). Hospital cost was observed to be similar (average cost, $5989 and $5977 per patient, respectively). The clinical outcomes were equivalent, with adjusted mortality rates for hospital-based attending physicians of 3.2% vs 3.9% for clinic-based attending physicians (P =.28). CONCLUSION: An increase of faculty time and involvement for supervision of resident-managed hospital care did not improve clinical outcomes or decrease costs during the 1-year study period.

Self-rated psychosocial consequences and quality of life in the acute porphyrias.

A battery of self-report psychosocial measures was mailed to 116 patients who had been referred for clinical management (clinic attenders) or laboratory diagnosis (non-clinic attenders) to the London Supraregional Assay Service Centre for Porphyria over the past decade and who tested positive for porphyria. Usable replies were received from 81 (70%) patients. Our interest focused on the prevalence of psychosocial symptoms in acute porphyrias and the perceived effects of porphyria on quality of life and patient experience. Research questions examined included (i), lifestyle factors; (ii) life events; (iii) mental health; (iv) general health; and (v) perceptions of illness of patients receiving specialist clinical management compared to respondents referred for diagnostic investigations, between patients with latent or manifest symptomology and between patients with different types of porphyria. Patients with porphyria have an impaired quality of life, particularly manifest cases, compared to controls and to diabetic patients. Depression, and particularly anxiety, is more common than in the general population or general medical outpatient attenders. Quality oflife is lower in acute intermittent porphyria (AIP) than in other forms of porphyria and a significant number of patients had major life event consequences, e.g. failure to secure, or loss of, employment, limitation of family size. Patients attending a clinic providing specialist porphyria advice, management and counselling received some perceived lifestyle benefits.

The atomic resolution structure of bucandin, a novel toxin isolated from the Malayan krait, determined by direct methods.

Bucandin is a novel presynaptic neurotoxin isolated from Bungarus candidus (Malayan krait). It has the unique property of enhancing presynaptic acetylcholine release and represents a family of three-finger toxins with an additional disulfide in the first loop. There are no existing structures from this sub-category of three-finger toxins. The X-ray crystal structure of bucandin has been determined by the Shake-and-Bake direct-methods procedure. The resulting electron-density maps were of outstanding quality and allowed the automated tracing of 61 of the 63 amino-acid residues, including their side chains, and the placement of 48 solvent molecules. The 0.97 A resolution full-matrix least-squares refinement converged to a crystallographic R factor of 12.4% and the final model contains 118 solvent molecules. This is the highest resolution structure of any member of the three-finger toxin family and thus it can serve as the best model for other members of the family. Furthermore, the structure of this novel toxin will help in understanding its unique ability to enhance acetylcholine release. The unique structure resulting from the fifth disulfide bond residing in the first loop improves the understanding of other toxins with a similar arrangement of disulfide bonds.

The crystal structure of ADP-L-glycero-D-mannoheptose 6-epimerase: catalysis with a twist.

BACKGROUND: ADP-L-glycero--mannoheptose 6-epimerase (AGME) is required for lipopolysaccharide (LPS) biosynthesis in most genera of pathogenic and non-pathogenic Gram-negative bacteria. It catalyzes the interconversion of ADP-D-glycero-D-mannoheptose and ADP-L-glycero-D-mannoheptose, a precursor of the seven-carbon sugar L-glycero-mannoheptose (heptose). Heptose is an obligatory component of the LPS core domain; its absence results in a truncated LPS structure resulting in susceptibility to hydrophobic antibiotics. Heptose is not found in mammalian cells, thus its biosynthetic pathway in bacteria presents a unique target for the design of novel antimicrobial agents. RESULTS: The structure of AGME, in complex with NADP and the catalytic inhibitor ADP-glucose, has been determined at 2.0 A resolution by multiwavelength anomalous diffraction (MAD) phasing methods. AGME is a homopentameric enzyme, which crystallizes with two pentamers in the asymmetric unit. The location of 70 crystallographically independent selenium sites was a key step in the structure determination process. Each monomer comprises two domains: a large N-terminal domain, consisting of a modified seven-stranded Rossmann fold that is associated with NADP binding; and a smaller alpha/beta C-terminal domain involved in substrate binding. CONCLUSIONS: The first structure of an LPS core biosynthetic enzyme leads to an understanding of the mechanism of the conversion between ADP-D-glycero--mannoheptose and ADP-L-glycero-D-mannoheptose. On the basis of its high structural similarity to UDP-galactose epimerase and the three-dimensional positions of the conserved residues Ser116, Tyr140 and Lys144, AGME was classified as a member of the short-chain dehydrogenase/reductase (SDR) superfamily. This study should prove useful in the design of mechanistic and structure-based inhibitors of the AGME catalyzed reaction.

Ill-conditioned Shake-and-Bake: the trap of the false minimum.

The alternation of phase refinement with the imposition of real-space constraints is the essence of the Shake-and-Bake procedure. Typically, these constraints prevent trial structures from falling into local minima. Nevertheless, P1 structures appear to migrate to false minima with significant frequency. These false minima are characterized by the presence of a large 'uranium' peak on the corresponding Fourier map. Fortunately, they can be recognized and avoided by considering the values of the minimal function both before and after the application of constraints. However, it appears that finding solutions for large P1 structures is likely also to require parameter-shift conditions different from those that have been found to work well in other space groups. In fact, these conditions often yield an unusually high percentage of solutions.