Joint action malnutrition in the elderly (MaNuEL) knowledge hub: summary of project findings.

PURPOSE: The Joint Action Malnutrition in the Elderly (MaNuEL) Knowledge Hub was established to extend scientific knowledge, strengthen evidence-based practice, build a sustainable, transnational network of experts and harmonize research and clinical practice in the field of protein-energy malnutrition in older persons. This paper aims to summarize the main scientific results achieved during the 2-year project and to outline the recommendations derived. METHODS: 22 research groups from seven countries (Austria, France, Germany, Ireland, Spain, The Netherlands and New Zealand) worked together on 6 relevant domains of malnutrition-i.e. prevalence, screening, determinants, treatment, policy measures and education for health care professionals-making use of existing datasets, evidence and expert knowledge. RESULTS: Four systematic reviews, six secondary data analyses of existing cohort and intervention studies, two web-based surveys and one Delphi study were performed. In addition, a scoring system to rate malnutrition screening tools and a theoretical framework on the aetiology of malnutrition in older persons were developed. Based on these activities and taking existing evidence into consideration, 13 clinical practice, 9 research and 4 policy recommendations were developed. The MaNuEL Toolbox was created and made available to effectively distribute and disseminate the MaNuEL results and recommendations. CONCLUSIONS: The MaNuEL Knowledge Hub successfully achieved its aims. Results and recommendations will support researchers, healthcare professionals, policy-makers as well as educational institutes to advance their efforts in tackling the increasing problem of protein-energy malnutrition in the older population.

Vitamin K status and inflammation are associated with cognition in older Irish adults.

Studies have shown associations between reduced vitamin K status and poor cognitive function. However, despite this apparent link, direct studies measuring cognitive function, vitamin K status and inflammation are lacking. In the current study, The ELDERMET cohort was investigated to identify associations between cognition, vitamin K status and inflammation. The primary aim of the ELDERMET study was to investigate the relationship between gut bacteria, diet, lifestyle and health in 500 older Irish adults. Significant differences in serum phylloquinone, dietary phylloquinone and inflammatory markers were found across varying levels of cognitive function, after controlling for sex, age, body mass index (BMI), triglycerides and blood pressure. In addition, significantly higher levels of dietary phylloquinone were found in those with better cognition compared to those with the poorest function. Higher levels of inflammatory were also associated with poor cognition. Furthermore, both dietary and serum phylloquinone were significant independent predictors of good cognitive function, after controlling for confounders. This study highlights the importance of dietary vitamin K as a potentially protective cognitive factor; it also provides evidence for the correlation between cognition and inflammation. Strategies should be devised by which elderly populations can access rich dietary sources of phylloquinone to maintain cognition.

Potentially modifiable determinants of malnutrition in older adults: A systematic review.

BACKGROUND & AIMS: Malnutrition in older adults results in significant personal, social, and economic burden. To combat this complex, multifactorial issue, evidence-based knowledge is needed on the modifiable determinants of malnutrition. Systematic reviews of prospective studies are lacking in this area; therefore, the aim of this systematic review was to investigate the modifiable determinants of malnutrition in older adults. METHODS: A systematic approach was taken to conduct this review. Eight databases were searched. Prospective cohort studies with participants of a mean age of 65 years or over were included. Studies were required to measure at least one determinant at baseline and malnutrition as outcome at follow-up. Study quality was assessed using a modified version of the Quality in Prognosis Studies (QUIPS) tool. Pooling of data in a meta-analysis was not possible therefore the findings of each study were synthesized narratively. A descriptive synthesis of studies was used to present results due the heterogeneity of population source and setting, definitions of determinants and outcomes. Consistency of findings was assessed using the schema: strong evidence, moderate evidence, low evidence, and conflicting evidence. RESULTS: Twenty-three studies were included in the final review. Thirty potentially modifiable determinants across seven domains (oral, psychosocial, medication and care, health, physical function, lifestyle, eating) were included. The majority of studies had a high risk of bias and were of a low quality. There is moderate evidence that hospitalisation, eating dependency, poor self-perceived health, poor physical function and poor appetite are determinants of malnutrition. Moderate evidence suggests that chewing difficulties, mouth pain, gum issues co-morbidity, visual and hearing impairments, smoking status, alcohol consumption and physical activity levels, complaints about taste of food and specific nutrient intake are not determinants of malnutrition. There is low evidence that loss of interest in life, access to meals and wheels, and modified texture diets are determinants of malnutrition. Furthermore, there is low evidence that psychological distress, anxiety, loneliness, access to transport and wellbeing, hunger and thirst are not determinants of malnutrition. There appears to be conflicting evidence that dental status, swallowing, cognitive function, depression, residential status, medication intake and/or polypharmacy, constipation, periodontal disease are determinants of malnutrition. CONCLUSION: There are multiple potentially modifiable determinants of malnutrition however strong robust evidence is lacking for the majority of determinants. Better prospective cohort studies are required. With an increasingly ageing population, targeting modifiable factors will be crucial to the effective treatment and prevention of malnutrition.

The Predictive Ability of Pre-Operative Magnetic Resonance Imaging to Detect Pathological Outcomes in Prostate Cancer

Aims Accurate preoperative knowledge of tumour stage is important in preoperative planning at radical prostatectomy (RP). The aim of this study was to assess the predictive ability of multiparametric MRI for detecting pathological outcomes. Methods A retrospective review was performed of all patients who underwent RP over a 4 year period. Results Preoperative MRI was reported as showing T3 or T4 disease in 26(17.9%) out of 145 patients undergoing RP. Of these, 10(6.9%) had ECE (extra-capsular extension) and 1(0.7%) had SVI (seminal vesicle invasion) on final histology. The sensitivity and specificity of MRI for detecting ECE were 27.3% and 87.6%, respectively. The sensitivity and specificity of MRI for detecting SVI were 11.1% and 97.8%, respectively. The positive predictive values for determining ECE and SVI were 45.5% and 25%, respectively and negative predictive values were 75.9% and 94.4%. Conclusion MRI has good specificity but poor and heterogeneous sensitivity for predicting T3 disease in RP specimen.

Plating up appropriate portion sizes for children: a systematic review of parental food and beverage portioning practices.

Consumption of larger portion sizes is associated with higher energy intake and weight status in children. As parents play a pivotal role in child feeding, we synthesized literature on 'parental portioning practices' using a mixed methods systematic design to inform future strategies addressing portion sizes served to children. Electronic databases PubMed, EMBASE, SCOPUS, PsycINFO and CINAHL Plus were searched. Two reviewers independently screened 385 abstracts and assessed 71 full-text articles against eligibility criteria: studies assessing portioning of foods or beverages by parent(s) with ≥1 child aged 2-12 years. Narrative synthesis of 14 quantitative studies found that portion sizes parents serve vary substantially and are influenced by amounts parents serve themselves, perceived child hunger and parent and child body size. Thematic synthesis of 14 qualitative studies found that parents serve the portion sizes they learn to be appropriate for their child to be fed. Portioning is influenced by parents' desires for a healthy child with a balanced diet. Future guidance on appropriate portion sizes for children would ideally present recommended portion sizes for first serving, incremental with age. Future research is however needed to assess the adoption and efficacy of providing such guidance to families.

Comparison of nomograms predicting lymph node invasion in patients undergoing radical prostatectomy for prostate cancer.

INTRODUCTION: The aim of this study was to compare the performance of preoperative risk nomograms or detecting lymph node invasion in a cohort of men undergoing radical prostatectomy (RP). METHODS: A retrospective analysis was performed on all men (n = 145) who underwent RP between 2012 and 2015. Preoperative data was inputted to the Memorial Sloan-Kettering Cancer Centre (MSKCC), Partin 2011 and Briganti nomograms and the University of California San Francisco- Centre of the Prostate Risk Assessment tool (UCSF-CAPRA). The risk of lymph node involvement (LNI) was calculated and compared to final histology. RESULTS: One hundred three (71%) men underwent a lymph node dissection at RP. Ten (9.7%) demonstrated LNI. The median nodal yield was 15 nodes, with no difference between those with LNI and those without (19.5 vs 14.5, p = 0.22).No patient classified as low risk on the UCSF-CAPRA score had evidence of LNI. In patients with LNI, no patient breached the 2% threshold for lymph node dissection (LND) on the MSKCC nomogram; four patients breached the 5% threshold on the Partin tables while three patients breached the 2.5% threshold for the Briganti nomogram. CONCLUSION: Nomograms produce useful information regarding risk of disease; however, they often have not been validated on different populations. Risk predictions need to be considered carefully and treatment decisions were made on a patient specific basis.

Ireland's contribution to urology and nephrology research in the new millennium: a bibliometric analysis.

INTRODUCTION: Bibliometrics is the statistical analysis of written publications. Bibliometric analyses have been performed across a range of biomedical disciplines. The aim of this study was to provide a comprehensive qualitative and quantitative analysis of Irish urology and nephrology research and to analyse how this compares internationally. METHODS: We performed a retrospective bibliometric analysis of the top 20 ranking journals in the field of "Urology and Nephrology" based on their 5 years impact factor, as obtained from the ISI Journal Citation Report database over the 15-year study period, 2000-2015. Utilising the Pubmed database, a search phrase was constructed using country of affiliation, year of publication and journal title. The abstracts of the Irish publications identified were analysed for their institution of origin, article theme and content. RESULTS: A total of 67,740 article abstracts were analysed over the 15 years study period. As anticipated, the USA accounted for the largest number of publications by a country [28,206 (41.64 % of all articles)]. Ireland contributed 347 articles in total (0.51 % of all articles); however, ranking according to population per million was 13th worldwide. Ireland's contribution to urology and nephrology research was highest in the BJUI-British Journal of Urology International [76 articles (21.90 % of Irish total)]. CONCLUSION: We believe this study to be the largest bibliometric analysis in the field of urology and nephrology internationally. This study provides a novel overview of the current Irish urology- and nephrology-related research, and examines how our results compare within the international community.

An Analysis of Gender Diversity in Urology in the UK and Ireland.

Traditionally, surgery and certain surgical sub-specialities in particular have been predominantly male orientated. In recent years, there has been an increased proportion of female medical graduates which will ultimately have an effect on speciality choices. The aim of this study was to assess the gender diversity among urologists in the UK and Ireland. The total number and gender breakdown of consultant urologists and trainees in the UK and Ireland was obtained from the British Association of Urological Surgeons (BAUS) and the Irish Society of Urology (ISU) membership offices. The total number and gender breakdown of medical school entrants and graduates in 2015 was obtained from the six medical schools in the Republic of Ireland. There are a total of 1,012 consultant urologists in the UK and Ireland. In the UK, 141 (14.6%) are female compared to four (8.2%) in Ireland, p= 0.531. There was a significant increase in the number of females between consultant urologists and trainees in both the UK (p=0.0001) and Ireland (p=0.015). In recent years, there has been a significant change in the percentage of female trainees in the UK and Ireland (22.8% (n=75) in 2011 vs 31.7% (n=93) in 2014, p=0.019. Between the six medical schools in Ireland, there were significantly more female entrants (n=726, 56.5%) than female graduates (n=521, 51.2%) in 2015, p=0.013.There has been a significant shift in gender diversity in urology in the UK and Ireland. Efforts to increase diversity should be pursued to attract further trainees to urology.

Uptake of selenium and mercury by captive mink: Results of a controlled feeding experiment.

Captive, juvenile, ranch-bred, male mink (Neovison vison) were fed diets containing various concentrations of methyl-mercury (MeHg) and selenium (Se) for a period of 13 weeks and then sacrificed to determine total Hg levels in fur, blood, brain, liver and kidneys and total Se concentrations in brain tissue. As MeHg concentrations in the diet increased, concentrations of total Hg in the tissues also increased with the highest level occurring in the fur > liver = kidney > brain > blood. Concentrations of Hg in the fur were correlated (r(2) > 0.97) with liver, kidney, blood and brain concentrations. The addition of Se to the mink diet did not appear to affect most tissue concentrations of total Hg nor did it affect the partitioning of Hg between the liver:blood, kidney:blood and brain:blood; however, partitioning of Hg between fur and blood was apparently affected.

Diet-microbiota-health interactions in older subjects: implications for healthy aging.

With modern medicine and an awareness of healthy lifestyle practices, people are living longer and generally healthier lives than their ancestors. These successes of modern medicine have resulted in an increasing proportion of elderly in society. Research groups around the world have investigated the contribution of gut microbial communities to human health and well-being. It was established that the microbiota composition of the human gut is modulated by lifestyle factors, especially diet. The microbiota composition and function, acting in concert with direct and indirect effects of habitual diet, is of great importance in remaining healthy and active. This is not a new concept, but until now the scale of the potential microbiota contribution was not appreciated. There are an estimated ten times more bacteria in an individual than human cells. The bacterial population is relatively stable in adults, but the age-related changes that occur later in life can have a negative impact on host health. This loss of the adult-associated microbiota correlates with measures of markers of inflammation, frailty, co-morbidity and nutritional status. This effect may be greater than that of diet or in some cases genetics alone. Collectively, the recent studies show the importance of the microbiota and associated metabolites in healthy aging and the importance of diet in its modulation.

Food and nutrient intake of Irish community-dwelling elderly subjects: who is at nutritional risk?

OBJECTIVES: To assess the dietary intakes of Irish community-dwelling elderly individuals, participating in the ELDERMET project. DESIGN: Cross-sectional study. SETTING: Cork city and county region of southern Ireland. PARTICIPANTS: Two hundred and eight (94 males, 114 females) community-dwelling subjects aged 64-93 yrs. MEASUREMENTS: Dietary intake was assessed using a validated semi-quantitative food frequency questionnaire (FFQ). Anthropometric data were recorded. Nutritional status was assessed using the Mini Nutritional Assessment (MNA). RESULTS: A high rate of overweight/obesity was observed in this population group. Consumption of energy-dense, low-nutrient foods was excessive among this population group. Older elderly subjects (≥75 yrs) consumed significantly (P<0.01) more desserts/sweets than younger elderly (64-74 yrs). Intakes of dietary fat and saturated fat were high while dairy food consumption was inadequate in both males and females. Elderly females typically had a more nutrient-dense diet than males. A considerable proportion of subjects, particularly males, had inadequate intakes of calcium, magnesium, vitamin D, folate, zinc and vitamin C. CONCLUSION: The data indicate that the diet of Irish community-dwelling elderly individuals is sub-optimal with respect to nutrient intake, and excessive in terms of fat intake, with implications for the health status of this population group. Reductions in dietary fat and increased low fat dairy food intakes are recommended for the prevention of diet-related disease in older persons. In addition, strategies to improve a number of sub-optimal micronutrient intakes need to be developed and implemented, particularly among elderly males.

The relationships among work stress, strain and self-reported errors in UK community pharmacy.

BACKGROUND: Changes in the UK community pharmacy profession including new contractual frameworks, expansion of services, and increasing levels of workload have prompted concerns about rising levels of workplace stress and overload. This has implications for pharmacist health and well-being and the occurrence of errors that pose a risk to patient safety. Despite these concerns being voiced in the profession, few studies have explored work stress in the community pharmacy context. OBJECTIVES: To investigate work-related stress among UK community pharmacists and to explore its relationships with pharmacists' psychological and physical well-being, and the occurrence of self-reported dispensing errors and detection of prescribing errors. METHOD: A cross-sectional postal survey of a random sample of practicing community pharmacists (n = 903) used ASSET (A Shortened Stress Evaluation Tool) and questions relating to self-reported involvement in errors. Stress data were compared to general working population norms, and regressed on well-being and self-reported errors. RESULTS: Analysis of the data revealed that pharmacists reported significantly higher levels of workplace stressors than the general working population, with concerns about work-life balance, the nature of the job, and work relationships being the most influential on health and well-being. Despite this, pharmacists were not found to report worse health than the general working population. Self-reported error involvement was linked to both high dispensing volume and being troubled by perceived overload (dispensing errors), and resources and communication (detection of prescribing errors). CONCLUSIONS: This study contributes to the literature by benchmarking community pharmacists' health and well-being, and investigating sources of stress using a quantitative approach. A further important contribution to the literature is the identification of a quantitative link between high workload and self-reported dispensing errors.

Modelling phosphorus dynamics in multi-branch river systems: a study of the Black River, Lake Simcoe, Ontario, Canada.

High rates of nutrient loading from agricultural and urban development have resulted in surface water eutrophication and groundwater contamination in regions of Ontario. In Lake Simcoe (Ontario, Canada), anthropogenic nutrient contributions have contributed to increased algal growth, low hypolimnetic oxygen concentrations, and impaired fish reproduction. An ambitious programme has been initiated to reduce phosphorus loads to the lake, aiming to achieve at least a 40% reduction in phosphorus loads by 2045. Achievement of this target necessitates effective remediation strategies, which will rely upon an improved understanding of controls on nutrient export from tributaries of Lake Simcoe as well as improved understanding of the importance of phosphorus cycling within the lake. In this paper, we describe a new model structure for the integrated dynamic and process-based model INCA-P, which allows fully-distributed applications, suited to branched river networks. We demonstrate application of this model to the Black River, a tributary of Lake Simcoe, and use INCA-P to simulate the fluxes of P entering the lake system, apportion phosphorus among different sources in the catchment, and explore future scenarios of land-use change and nutrient management to identify high priority sites for implementation of watershed best management practises.

Halocins and sulfolobicins: the emerging story of archaeal protein and peptide antibiotics.

Production of antibiotic peptides and proteins is a near-universal feature of living organisms regardless of phylogenetic classification. Bacteriocins (proteinaceous antimicrobials from the domain Bacteria) have been studied for over 75 years, and the eucaryocins (proteinaceous antimicrobials from the domain Eucarya) since the early 1960s. However, one domain of organisms, the Archaea, containing hyperthermophiles, extreme halophiles and the methanogens, is just beginning to be scrutinized for the production of peptide antibiotics. Production of archaeal proteinaceous antimicrobials (archaeocins) from extreme halophiles (halocins) is a nearly universal feature of the rod-shaped haloarchaea. Halocin activity is first detectable in culture supernatants at the beginning of the transition into stationary phase, concomitant with an induction of transcription of the structural gene. Halocins are diverse in size, consisting of proteins as large as 35 kDa and peptide "microhalocins" as small as 3.6 kDa. The 36 amino acids of microhalocin HalS8 are located in the interior of a 311-residue pro-protein from which they are liberated by an unknown mechanism. Microhalocins are hydrophobic and robust, withstanding heat, desalting and exposure to organic solvents. Unlike the peptide bacteriocins and the eucaryocins, microhalocins possess a large number of neutral residues and are not cationic, leaving their mechanism(s) of action mostly a mystery. While microhalocins affect a variety of haloarchaeal genera (kingdom Euryarchaeota), they also exhibit cross-kingdom toxicity, inhibiting or killing Sulfolobus species (kingdom Crenarchaeota). Finally, archaeocins also are produced by the hyperthermophile "Sulfolobus islandicus". These 20-kDa protein antibiotics are not excreted into the environment, but are associated with small particles apparently derived from the cell's S-layer.

Gender differences in 24-hour outcome following resuscitation after 9 minutes of cardiac arrest in dogs.

OBJECTIVE: To examine possible gender-specific differences in 24-hr outcome following resuscitation from 9 mins of controlled cardiac arrest. DESIGN: Preclinical, prospective study comparing two similarly prepared, independent control groups (one female group, one male group) included in a larger series of studies. SETTING: Physiology research laboratory at a major medical center. SUBJECTS: Male and female mongrel dogs (Canis familiaris), weighing 16 to 22 kg. INTERVENTIONS: Cardiopulmonary-cerebral resuscitation following 9 mins of normothermic cardiac arrest in male vs. female dogs. MEASUREMENTS AND MAIN RESULTS: Mean arterial blood pressure, heart rate, urine output, arterial blood oxygen, and PCO2 values, arterial pH, temperature, plasma glucose concentrations, and hematocrit were measured and recorded at the precardiac arrest and postcardiac arrest period, and at 30 mins, and 1, 4, 6, 12, and 24 hrs following resuscitation. Neurologic dysfunction was assessed using a well-standardized neurologic deficit score assigned at 6, 12, and 24 hrs after arrest. Plasma concentrations of malonaldehyde, 4-hydroxynonenal, and erythrocyte-reduced glutathione were measured at the precardiac arrest period, and 6, 12, and 24 hrs following resuscitation. Additionally, serum concentrations of alanine aminotransferase, aspartate aminotransferase, total bilirubin, alkaline phosphatase, gamma-glutamyl transferase, creatinine kinase, creatinine, albumin, and total protein were measured before arrest, and at 6, 12, and 24 hrs after resuscitation. Plasma concentrations of inorganic phosphorus, blood urea nitrogen, and electrolytes (sodium, chloride, calcium, and potassium) were measured. The estrous cycle phase in the female dogs enrolled in the study was determined by physical examination and vaginal cytology. No prearrest differences were detectable between males and females in basic physiologic variables. No differences in neurologic deficit were detectable between males and females across the 24-hr recovery period following resuscitation. No detectable differences in malonaldehyde, 4-hydroxynonenal, and erythrocyte-reduced glutathione occurred between groups. Serum concentrations of aspartate aminotransferase (p = .02), alanine aminotransferase (p = .009), creatinine kinase (p = .01), total bilirubin (p = .05), and plasma concentrations of inorganic phosphorus (p = .03), blood urea nitrogen (p = .0003), and creatinine (p = .02) all were significantly and dramatically higher in female than male dogs at the 24-hr time point. The trend of increase in these values began at the 6- and 12-hr time points and was consistent with a steadily decreasing trend in mean arterial pressure and an increasing trend in heart rate in the female group. CONCLUSIONS: An extensive history with this preclinical canine model (restricted to male dogs) had indicated little or no change in standard clinical chemistry markers of systemic dysfunction following 9 mins of cardiac arrest. However, when compared with male dogs, the female dogs tested here appear to have sustained a more significant hepatic and renal ischemic injury with no differences in the neurologic deficit.

Isolation, sequence, and expression of the gene encoding halocin H4, a bacteriocin from the halophilic archaeon Haloferax mediterranei R4.

The first gene to encode a haloarchaeal bacteriocin (halocin H4) has been cloned and sequenced from Haloferax mediterranei R4. Both the signal sequence in the halocin H4 preprotein and the monocistronic halH4 gene have some unusual features. The physiology of halH4 expression reveals that although halH4 transcripts are present at low basal levels during exponential growth, halocin H4 activity first appears as the culture enters stationary phase. As halocin activity levels increase, so do transcript levels, but then activity levels decrease precipitously while transcript levels remain elevated.

Thermal strain and G protection associated with wearing an enhanced anti-G protection system in a warm climate.

A flight trial was conducted in Cyprus to assess the thermal strain associated with and the G protection provided by the prototype Eurofighter 2000 aircrew equipment assembly (AEA) in a warm climate. Six subjects flew a standardized sortie four times in a Hawk aircraft: two while wearing the Eurofighter 2000 AEA and two wearing standard Hawk summer AEA. The sortie included high-G turns and simulated air combat. Cockpit temperatures, rectal and skin temperatures, heart rate, and sweat rate were recorded. Subjective thermal comfort, fatigue, and G protection were also assessed. Skin temperatures of the back, chest and thighs, mean skin temperatures, and sweat rate were greater when the Eurofighter AEA was worn. Rectal temperature and heart rate did not differ significantly between the two conditions. Superior G protection was provided by the Eurofighter assembly. These findings suggest that wearing the Eurofighter AEA in a warm climate is associated with an increased but not unacceptable level of thermal stress while offering enhanced G protection. These results may not generalize when ambient temperatures are higher or more insulative protective clothing is worn.

Immunohistochemical localization of smooth muscle myosin in human spleen, lymph node, and other lymphoid tissues. Unique staining patterns in splenic white pulp and sinuses, lymphoid follicles, and certain vasculature, with ultrastructural correlations.

The anatomic distribution of smooth muscle myosin, a contractile protein, was determined in a variety of lymphoid tissues (spleen, lymph nodes, tonsils) with the use of highly specific rabbit antibodies to human uterine smooth muscle myosin and an indirect immunoperoxidase technique. In the spleen, in addition to the anticipated immunoreactivity in the walls of arteries, veins, splenic capsule, and trabeculas, other staining patterns were observed. Smooth muscle myosin-containing cells which comprised the adventitia of the trabecular arteries appeared continuous with myosin-containing reticular cells of the white pulp. The latter cells assumed a circumferential pattern within the periarteriolar lymphoid sheaths, then blended delicately with the red pulp at the marginal zone. Ultrastructurally, immunogold techniques demonstrated that smooth muscle myosin in these cells was localized to cytoplasmic filaments. Within the red pulp, a different and distinct staining pattern was observed for the splenic sinuses. Short, regular, orderly, and repetitive bands of immunoreactivity, aligned parallel to the long axis of the sinus, extended between contiguous ring fibers. By immunoelectron microscopy these structures corresponded to distinct bundles of filaments in the endothelial lining cells of the splenic sinuses. Factor VIII associated antigen was also identified in the splenic lining cells in cryostat and paraffin sections, and ultrastructurally. Within the red pulp of the spleen, the sheaths of sheathed capillaries also revealed strong immunoreactivity for smooth muscle myosin. Other sites of immunohistochemical localization of smooth muscle myosin included dendritic reticulum cells present in reactive follicles and in nodular non-Hodgkin's lymphomas. Certain vascular structures, specifically sinus lining cells and Schweigger-Seidel capillary sheaths of the spleen and postcapillary venules of lymph nodes and tonsils, coexpressed smooth muscle myosin and Factor VIII associated antigen. The patterns of localization of smooth muscle myosin are correlated with anatomic structures and possible tissue functions.

Are keratin proteins a better tumor marker than epithelial membrane antigen? A comparative immunohistochemical study of various paraffin-embedded neoplasms using monoclonal and polyclonal antibodies.

Epithelial membrane antigen and keratin proteins represent markers of epithelial differentiation that may be detected in routine formalin-fixed, paraffin-embedded tissues. Eighty-seven neoplasms, including 48 adenocarcinomas of various types, squamous and transitional cell carcinomas, small-cell anaplastic carcinomas, carcinoid tumors, mesotheliomas, hepatomas, melanomas (metastatic), adrenal cortical carcinomas, germ cell tumors, and extramammary Paget's disease, were assessed to determine the relative effectiveness of these antigens as tumor markers. Immunoperoxidase studies were performed using monoclonal antibodies to epithelial membrane antigen and monoclonal (combined AE1 and AE3) and polyclonal (bovine muzzle keratins) antibodies to keratin proteins. In more than half the cases (50/87%), both markers yielded comparable results. However, in 29 cases (33%), keratin proteins were clearly superior to epithelial membrane antigen as a tumor cell marker. Particular discrepancies were apparent for some gastrointestinal adenocarcinomas, squamous cell carcinomas, hepatomas (hepatocellular type), spindle cell components of mesotheliomas, and carcinoid tumors. Epithelial membrane antigen represented a better marker in eight cases (9%), mainly for small-cell anaplastic carcinomas and some renal cell and pulmonary adenocarcinomas. Adrenal cortical carcinomas, melanomas, and seminomas were nonimmunoreactive for both antigens. Epithelial membrane antigen and keratin proteins represent useful complementary markers in diagnostic surgical pathology.

Optimal immunoreactivity of keratin proteins in formalin-fixed, paraffin-embedded tissue requires preliminary trypsinization. An immunoperoxidase study of various tumours using polyclonal and monoclonal antibodies.

The effect of preliminary trypsinization on the immunoreactivity of keratin proteins in formalin-fixed, paraffin-embedded tissues of a variety of tumors (squamous cell carcinomas, adenocarcinomas, mesotheliomas, and transitional cell carcinomas) was evaluated. Three types of trypsin (Type II and Type IX porcine trypsin and Type III bovine trypsin) and varying concentrations of trypsin were assessed. Immunoreactivity of keratin proteins was determined using rabbit anti-keratin antibodies and monoclonal antibodies (combination of AE1 and AE3) and immunoperoxidase techniques. Preliminary trypsinization was mandatory for optimal immunoreactivity of keratin proteins using either polyclonal or monoclonal antibodies. Excellent results were obtained using Type II porcine trypsin at concentrations of 25 mg/dl for 30-45 min or 50 mg/dl for 20 min, at 37 degrees C. Trypsin treatment with excessive concentrations of enzyme and/or extended incubation times promoted tissue digestion and in some cases, yielded decreased immunoreactivity and altered staining patterns.