Palaeoecological data indicates land-use changes across Europe linked to spatial heterogeneity in mortality during the Black Death pandemic.

The Black Death (1347-1352 CE) is the most renowned pandemic in human history, believed by many to have killed half of Europe's population. However, despite advances in ancient DNA research that conclusively identified the pandemic's causative agent (bacterium Yersinia pestis), our knowledge of the Black Death remains limited, based primarily on qualitative remarks in medieval written sources available for some areas of Western Europe. Here, we remedy this situation by applying a pioneering new approach, 'big data palaeoecology', which, starting from palynological data, evaluates the scale of the Black Death's mortality on a regional scale across Europe. We collected pollen data on landscape change from 261 radiocarbon-dated coring sites (lakes and wetlands) located across 19 modern-day European countries. We used two independent methods of analysis to evaluate whether the changes we see in the landscape at the time of the Black Death agree with the hypothesis that a large portion of the population, upwards of half, died within a few years in the 21 historical regions we studied. While we can confirm that the Black Death had a devastating impact in some regions, we found that it had negligible or no impact in others. These inter-regional differences in the Black Death's mortality across Europe demonstrate the significance of cultural, ecological, economic, societal and climatic factors that mediated the dissemination and impact of the disease. The complex interplay of these factors, along with the historical ecology of plague, should be a focus of future research on historical pandemics.

Understanding and predicting the impact of critical dissolution variables for nifedipine immediate release capsules by multivariate data analysis.

In this study the selection of in vivo predictive in vitro dissolution experimental set-ups using a multivariate analysis approach, in line with the Quality by Design (QbD) principles, is explored. The dissolution variables selected using a design of experiments (DoE) were the dissolution apparatus [USP1 apparatus (basket) and USP2 apparatus (paddle)], the rotational speed of the basket/or paddle, the operator conditions (dissolution apparatus brand and operator), the volume, the pH, and the ethanol content of the dissolution medium. The dissolution profiles of two nifedipine capsules (poorly soluble compound), under conditions mimicking the intake of the capsules with i. water, ii. orange juice and iii. an alcoholic drink (orange juice and ethanol) were analysed using multiple linear regression (MLR). Optimised dissolution set-ups, generated based on the mathematical model obtained via MLR, were used to build predicted in vitro-in vivo correlations (IVIVC). IVIVC could be achieved using physiologically relevant in vitro conditions mimicking the intake of the capsules with an alcoholic drink (orange juice and ethanol). The multivariate analysis revealed that the concentration of ethanol used in the in vitro dissolution experiments (47% v/v) can be lowered to less than 20% v/v, reflecting recently found physiological conditions.

In vitro and in silico characterisation of Tacrolimus released under biorelevant conditions.

This work aims to better understand the in vivo behaviour of modified release (MR) formulations (Envarsus® tablets and Advagraf® capsules) using in vitro properties of tacrolimus and in silico simulations. The in silico concentration profiles of tacrolimus released from the MR formulations were predicted after building a three compartments PK model with GastroPlus™, and using the experimentally determined in vitro physico-chemical properties as input parameters. In vitro-in vivo correlations (IVIVC) were obtained after deconvolution of in vivo data from a clinical trial. The IVIVC showed that the in vitro dissolution was faster than the in vivo deconvoluted dissolution for Advagraf®, while the in vitro dissolution was slightly slower than the in vivo deconvoluted dissolution for Envarsus®. Population PK simulation showed that variability in the simulation was lower for Envarsus® compared to Advagraf®. The in silico predicted preferential absorption sites were the proximal and distal tract for Advagraf® and Envarsus®, respectively. The integration of experimental in vitro solubility, permeability and biorelevant dissolution data allowed to generate in silico tacrolimus concentrations for two different MR formulations. This permitted to compare the two formulations in a single PK profile, in a simulated population PK study and with respect to their absorption sites.

An in vitro-in vivo correlation study for nifedipine immediate release capsules administered with water, alcoholic and non-alcoholic beverages: Impact of in vitro dissolution media and hydrodynamics.

The impact of hydrodynamics and media composition on nifedipine dissolution profile from IR (immediate release) soft capsules was investigated using dissolution apparatus USP1, USP2, USP3 and USP4 (United State Pharmacopoeia). Media composition was varied in terms of pH and content, to mimic the dosage form intake with water or non-alcoholic beverages (orange juice) and alcoholic beverages (orange juice/ethanol mixture (47% v/v)). Through construction of in vitro-in vivo correlations (IVIVC) with corresponding in vivo data from the literature, it was possible to evaluate the in vitro conditions that are likely to simulate the in vivo formulation behaviour. Both linear and nonlinear correlations were obtained depending on experimental set-ups. Testing of 20mg nifedipine capsules in FaSSGFst (Fasted State Simulated Gastric Fluid pH 1.6; water administration) produced IVIVC with the USP3 (after time scaling) and USP4 apparatus. IVIVC were obtained for USP2, USP3 and USP4 in FaSSGFoj (Fasted State Simulated Gastric Fluid pH 3.4; orange juice administration). Linear and nonlinear correlations were obtained with the USP1, USP2 and USP3 apparatus when testing the capsules in FaSSGFoj/EtOH (orange juice/ethanol administration). This study highlighted that selection of physiologically relevant dissolution set-ups is critical for predicting the in vivo impact of formulations co-administration with water, non-alcoholic and alcoholic beverages.

Climate warming and the decline of Taxus airborne pollen in urban pollen rain (Emilia Romagna, northern Italy).

Woody plant performance in a changing global environment has always been at the centre of palaeoenvironmental and long-term climate reconstructions carried out by means of pollen analysis. In Mediterranean regions, Taxus constitutes the highest percentage in past pollen diagrams from cold or cool periods, and therefore it is generally considered a good index to infer climate features from past records. However, a comparison of these inferences with the true current trends in pollen production has not been attemped until now. This study reports the decline of airborne pollen of Taxus observed in Emilia Romagna, a region of northern Italy, during the period 1990-2007. Phenological observations on four male specimens and microscopic examination of fresh pollen were made in order to check Taxus flowering time and pollen morphology. Airborne pollen was monitored through continuous sampling with a Hirst volumetric sampler. In the 18-year long period of investigation, Taxus pollen production has decreased, while total woody pollen abundance in air has increased. The trend of the Taxus pollen season shows a delay at the beginning, a shortening of the pollen period, and an advance of the end of the pollen season. This was interpreted as a response to climate warming. In particular, Taxus follows the behaviour of winter-flowering plants, and therefore earlier pollination is favoured at low autumn temperatures, while late pollination occurs more often, most likely after warm autumn temperatures.

Identification and molecular interpretation of the effects of drug incorporation on the self-emulsification process using spectroscopic, micropolarimetric and microscopic measurements.

Addition of a drug to a self-emulsifying drug delivery system (SEDDS) can affect the emulsification process after administration, leading to variation in the emulsion droplet size formed and potentially its clinical behavior (Mercuri et al., Pharm. Res., 2011, 28, 1540-1551). However, the mechanisms involved and, in particular, the location of the drug within the system are poorly understood. Here, we have investigated the location of a model drug, ibuprofen, in the emulsions formed from a simple anhydrous SEDDS (soybean oil, Tween 80 and Span 80), using a range of physical characterization techniques. (1)H NMR studies showed an interaction between the drug and the polyoxyethylene chains of the surfactant Tween 80. Micropolarity assessment of the emulsion droplet interfacial region, using the chemical probes pyrene and Reichardt's dye, confirmed this interaction, and suggested that the drug was altering the microenvironment around the surfactants, and hence the behavior of the SEDDS with water during emulsification. Both dielectric spectroscopy and polarized light microscopy highlighted the differential behavior with water of placebo and drug-loaded SEDDS, also seen in the initial visual observational studies on the emulsification performance of the SEDDS. (1)H NMR studies with three other NSAIDs indicate that this effect is not confined to ibuprofen alone. The study has therefore indicated that the drug's influence on the emulsification process may be related to interactions within the microenvironment of the surfactant layer. Furthermore, such interactions may be usefully identified and characterized using a combination of micropolarity, spectroscopic and microscopic methods.

Trends and challenges in tuberculosis in a medium-sized southern European setting.

Trends in tuberculosis (TB) admissions over 40 years at the Infectious Diseases Clinic of Perugia University Hospital, Perugia, Italy, show that in the last decade non-Italian TB case admissions outweighed those of Italians, with a large number of cases from Eastern Europe (25.2%) and Africa (23.4%). Non-Italians tended to be younger and were generally new pulmonary TB cases, and drug resistance was also more common. Overall, the number of multidrug-resistant cases increased. Only one case occurred in a native-born Italian, and five of seven cases had newly diagnosed TB. In low TB incidence settings such as Perugia, Italy, TB prevention and control programmes for the foreign-born need to be reinforced.

Predicting the human in vivo performance of different oral capsule shell types using a novel in vitro dynamic gastric model.

The disintegration of a capsule shell may determine the onset of drug dissolution from capsule formulations. In this study, the release of a rapidly dissolving model drug (paracetamol), from two hydroxypropyl methylcellulose capsules containing either carageenan (HPMC-C) or gellan gum (HPMC-G) and one hard gelatin (HG) capsule, were investigated using a conventional in vitro model, the USP dissolution apparatus I, and a novel in vitro model of the human gastric compartment, the dynamic gastric model (DGM). The results obtained in vitro were compared with in vivo gamma scintigraphy human data and in vivo gastric emptying profiles available in the literature. The drug release from HPMC-G capsules, observed with the USP dissolution apparatus I, was delayed with respect to the other two capsules, while the results obtained from the DGM in the fasted state were closer together, which was in agreement with data from the in vivo studies. In the fasted state, the capsule rupture times obtained from the DGM were similar to those observed by gamma scintigraphy in vivo studies. In the fed state, the 'apparent' rupture times observed with the DGM were delayed compared to fasted, and were even longer than those observed by scintigraphy in vivo for HPMC-G and HG capsules. However, these discrepancies can reasonably be explained by considering the impact of food upon dispersion of the capsule contents and the sampling from the DGM, when compared to the human scintigraphy experiments.

The utility of multimodality imaging with CT and MRI in defining rectal tumour volumes for radiotherapy treatment planning: a pilot study.

AIMS: This study compares the volumetric and spatial relationships of gross tumour volume (GTV) derived from CT (CT-GTV) and GTV derived from MRI (MR-GTV) to determine the utility of multi-modality imaging for radiotherapy treatment planning in rectal cancer. METHODS AND MATERIALS: Fifteen patients with T3 rectal cancer were accrued over 18 months. The male : female ratio was 2:1. The average age was 60.3 years (range 38-79). All patients underwent a diagnostic MRI and CT and MRI simulation. Data sets were co-registered. A site specialised diagnostic radiologist contoured all volumes in consultation with a radiation oncologist. CT-GTV was contoured while blinded to MR data sets. MR-GTV was contoured independently 2-4 weeks later whilst blinded to its respective CT-GTV data. Tumour volumes were analysed for three anatomical subregions (sigmoid, rectal and anal). Reference points on tumour volumes were used for spatial comparison and analysis. RESULTS: The mean CT-GTV/MR-GTV ratio was 1.2 (range 0.5-2.9). The tumour volume ratios for the rectal subregion were well correlated. CT-GTV provided adequate spatial coverage of tumour in reference to MR-GTV with the average mean discrepancy of 0.12 (range -0.08-0.38) or a maximum discrepancy of <0.4 cm (1.54 standard deviation). CT-GTV coverage was inadequate for tumours with MRI evidence of anal and sigmoid invasion. CONCLUSIONS: Conventional simulation CT imaging provided a reasonable estimate of the GTV. Multi-modality imaging with staging MRI can assist target volume definition where there is involvement of the sigmoid and anorectal region and avoid geographic misses. The role of a simulation MRI may aid in this process but remains investigational.

The effect of an intensive nutritional program on daily set-up variations and radiotherapy planning margins of head and neck cancer patients.

This is a prospective case-control study to assess nutritional supplementation in limiting weight loss and its impact on daily set-up variations and planning target volume (PTV) margins in head and neck (H&N) radiotherapy (RT). Twenty sequential H&N patients were recruited for this study. Ten patients had a percutaneous endoscopic gastrostomy (PEG) tube inserted prior to RT and 10 did not. PEG use was determined by departmental guidelines for patients considered at high risk for weight loss. Daily 2D electronic portal images were taken for orthogonal verification. Set-up variations were determined for both PEG and non-PEG patients by calculating systematic (Sigma) and random (sigma) errors, and PTV margins were derived. PEG patients lost less weight (P = 0.04) over the course of RT and had a reduction in set-up variation in the superior-inferior (SI) and anterior-posterior (AP) planes compared to those without. Mean correctional shifts in mm (range) for PEG patients were: Right-Left (RL) 0.1 (-1.9-2.1), SI -1.7 (-2.9-0.0), AP -0.4 (-2.0-0.8), and for non-PEG patients were: RL -0.2 (-2.7-1.3), SI -1.3 (-3.1-1.0), AP 0.4 (-1.5-2.8). The adapted PTV margins (mm) in the RL, SI and AP planes, respectively, for PEG patients were 4.1, 3.3 and 3.6, and for non-PEG were 3.9, 4.9 and 4.8. Intensive enteral support maintained weight stability in H&N patients considered at risk of weight loss during RT and this was associated with reduced set-up variation.

Development of a bioreactor system for the decolorization of dyes by Coriolus versicolor f. antarcticus.

Decolorization of 100 microM malachite green (MG) by Coriolus versicolor f. antarcticus using a two-phase bioreactor, was investigated. In the first phase the decolorization ability of this fungus, growing under conditions of solid-state fermentation (SSF), was proved; in the second phase the capacity of the enzymes present in extracts from the solid residues was exploited. During the first phase using the same culture in the bioreactor, five consecutive charges were made, each with 75 ml of 100 microM MG solution, at 28 degrees C. Each cycle ended when MG solution reached a decolorization of 50%, at this time the bioreactor was discharged to a stainless steel coil at 50 degrees C, initiating the second phase of decolorization. Time required in order to reach 50% decolorization during the first phase varied between 25 and 65 min, with an average retention time of 48 min. The second stage had a retention time of 120 min. Residual MG after this phase varied from 0% to 6.3%. The role of laccase and Mn-peroxidase in MG decolorization is discussed. Toxicity of MG solutions before and after decolorization treatments was assayed using Lumbriculus variegatus as test organism.

Vasoactive biomarkers and oxidative stress in healthy recently postmenopausal women treated with hormone replacement therapy.

BACKGROUND: Despite biologically plausible mechanisms for cardiac protection from estrogen therapy, recent clinical trials have suggested possible cardiovascular risk rather than benefit. However, it has been speculated that cardioprotective benefits from hormone replacement therapy (HRT) may be more evident in the early postmenopausal period. We have previously reported early beneficial effects on biochemical markers of endothelial function in healthy women after short-term estradiol replacement therapy. In this study we aimed to evaluate the effect of long-term HRT on different vasoactive factors and oxidative stress in healthy recently postmenopausal women. METHODS: Fifteen women (age 50 +/- 1 years, time since menopause 1.6 +/- 0.1 years) were randomized to a sequential oral and transdermal estradiol regimen (2 mg oral micronized 17beta-estradiol/day or 1.5 mg 17beta-estradiol gel/day). Oral dydrogesterone (10 mg/day, 12 days/month) was then cyclically combined with either of the estrogen therapies for 1 year. Blood samples were collected at baseline and after 1, 2, 6 and 12 months of therapy to evaluate levels of follicle stimulating hormone (FSH), estradiol, 6-keto PGF1alpha (prostacyclin metabolite), nitrite/nitrate, epinephrine, norepinephrine, 8-isoprostane (8-epi PGF2alpha) and lipid profile values. RESULTS: FSH levels decreased (p < 0.001) while estradiol levels increased (p < 0.001) during HRT. Levels of epinephrine (p < 0.001), norepinephrine (p < 0.01), mean blood pressure (p < 0.01) and low density lipoprotein (LDL) cholesterol (p < 0.01) decreased, and nitrite/nitrate levels increased (p < 0.01) during HRT, which did not significantly affect 8-epi PGF2alpha levels. CONCLUSIONS: One-year HRT significantly reduced the levels of catecholamines, mean blood pressure and LDL cholesterol while it increased levels of nitrite/nitrate, indicating cardiovascular benefit in healthy recent postmenopausal women. Levels of 8-epi PGF2alpha did not change, suggesting no evident relationship between HRT and oxidative stress.

Estimating the unknown components of nutrient mass balances for forestry plantations in mine rehabilitation, upper Hunter Valley, New South Wales, Australia.

Commercial forestry plantations as a postmining land use in the Upper Hunter Valley of New South Wales, Australia are restricted by both the poor nutrient availability of mining substrates and low regional rainfall. An experiment was conducted to investigate whether municipal waste products and saline groundwater from coal mining operations could improve early tree growth without impacting on the environment through salt accumulation and/or nutrient enrichment and changes in groundwater quality. Potential impacts were investigated by quantifying the nutrient cycling dynamics within the plantation using an input-output mass balance approach for exchangeable calcium (Ca(2+)), exchangeable magnesium (Mg(2+)), exchangeable potassium (K(+)), exchangeable sodium (Na(+)), nitrogen (N), and phosphorus (P). Measured inputs to and outputs from the available nutrient pool in the 0-30 cm of the overburden subsystem were used to estimate the net effect of unmeasured inputs and outputs (termed "residuals"). Residual values in the mass balance of the irrigated treatments demonstrated large leaching losses of exchangeable Ca, Mg, K, and Na. Between 96% and 103% of Na applied in saline mine-water irrigation was leached below the 0-30-cm soil profile zone. The fate of these salts beyond 30 cm is unknown, but results suggest that irrigation with saline mine water had minimal impact on the substrate to 30 cm over the first 2 years since plantation establishment. Accumulations of N and P were detected for the substrate amendments, suggesting that organic amendments (particularly compost) retained the applied nutrients with very little associated losses, particularly through leaching.

Effects of aerobic training on lactate and catecholaminergic exercise responses in mitochondrial myopathies.

The aim of this study was to evaluate the effects of an aerobic training program on the metabolic and sympathetic responses to exercise in 12 patients with mitochondrial myopathies. A 10-week course of aerobic training, consisting of supervised exercise every other day on an electrically braked pedal-rate bicycle ergometer was prescribed to each patient and four healthy controls. Venous lactate, epinephrine (EP) and norepinephrine (NEP) levels were assessed at baseline and after the aerobic training by means of constant-workload exercise performed at near lactate threshold (LT). In patients, a decrease in exercise peak values, significant for lactate (-38.6%, P < 0.01) but not for catecholamines (EP: -26.0%, NEP: -22.1%) was observed after training, findings confirmed by the lactate/EP and lactate/NEP area ratios. The results show that lactate accumulation during exercise is decreased after aerobic training in mitochondrial myopathies and that the effect is partially dissociated from the catecholaminergic response. This in turn suggests that the lactate decrease can be explained, at least in part, by the improved muscle oxidative metabolism consequent to the proposed training program.

[Registry of genetic and malformative diseases in the province of Macerata]. / Registro delle malattie genetiche e malformative della provincia di Macerata.

OBJECTIVES: The aim of this epidemiological research is to evaluate the prevalence of genetic diseases and malformative syndromes in paediatric population living in the Macerata county. MATERIAL AND METHODS: All the data were collected through a careful analysis of a specific questionnaire sent to all the family paediatricians. RESULTS: 23,379 children living in Macerata county, aged 0 to 9 years, were evaluated (93.8% of all this paediatric population). Among those were found N 400 cases of genetic diseases and malformative syndromes: Malformations Tot.N. 255 cases (63.3% of the reported cases); Malformative Syndromes Tot. N. 55 cases (27.8% of the reported cases); Endocrinology and Metabolic Diseases Tot. N. 41 cases (10.3% of the reported cases); Osteochondrodysplasia Tot. N. 22 cases (5.7% of the reported cases); Other Tot. N. 28 cases (7.0% of the reported cases); Male population was found more affected than female: M/F ratio = 1.4. The analysis of the data showed an increasing trend in detecting these pathological conditions, consistent with the increase in geographic altitude (3 areas considered): 0-100 meter = 0.88%; 100-600 m.a.s. = 1.34%; over 600 m.a.s. = 1.88%. CONCLUSION: The knowledge of the number of children affected by genetic and malformative diseases in the Macerata county is relevant in order to establish a Genetic Service with the aim to better support the medical assistance of these patients and counselling service for the families.

Aerobiologia 2.0: a software program for processing aeropollen data.

"Aerobiologia 2.0" is a simple computer program created to handle the pollen data collected every 2 hrs and daily by aerobiological monitoring stations equipped with Hirst-type spore traps. "Aerobiologia 2.0" runs on Windows 3.1 and is compatible with other programs that run on this operating system. The program was developed to store and process pollen data through a few straightforward operations. An unlimited calendar automatically calculates the day of the week. The pollen dictionary, which can hold up to 1216 different pollen types, may be modified or changed completely. Concentrations for every pollen type (in pollen grains/m3) are automatically recorded daily and every 2 hrs. 10-day and monthly sums are also calculated. The percentage of selected types, groups, or families of pollen collected each day, every 10 days, and monthly is quickly available. Pollen calendars and spectra in 24-hr, 10-day, monthly, tri-monthly, half-year, and yearly periods are readily produced. As soon as it is entered, the pollen data are saved on hard disk. A year's worth of data can be saved on a single 1.44 M byte floppy disk. Aerobiologia 2.0 is being used successfully to process the aeropollen data collected at the two monitoring stations managed by our Palynological Laboratory.

Child with oral, facial, digital, and skeletal anomalies and psychomotor delay: a new OFDS form?

We report on a male infant with oral, facial, digital, and skeletal anomalies in association with severe psychomotor delay. This may represent a new oral-facial-digital syndrome.

Performance of a fully automated fluorometric enzyme immunoassay for serum myoglobin measurement.

An automated fluorometric enzyme immunoassay system for the determination of serum myoglobin has been recently developed. This method is based on the sandwich immunoassay and uses two mouse monoclonal antimyoglobin antibodies; the first one is complexed onto glass fiber paper and the second is conjugated to an enzyme alkaline phosphatase which reacts with the substrate 4-methylumbelliferyl phosphate to generate a fluorescent product. Using a dedicated automated apparatus the time to the first result is eight minutes, with additional values being produced at one-minute intervals (about 50 samples/hour). We compared the analytical performance of this fluorometric enzyme immunoassay with that of a RIA set up in our laboratory for the routine assay of serum myoglobin. The automated fluorometric enzyme immunoassay showed lower between-assay variability (CV = 4.7% vs 13.8%) and higher sensitivity (0.3 ng/mL vs 7.2 ng/mL) than the manual RIA. Moreover, the two immunoassays gave similar results when serum samples of normal subjects and patients with coronary artery disease with or without acute myocardial infarction (AMI) were assayed (fluorometric immunoassay = -0.7 + 0.851 RIA, r = 0.991, n = 137). In conclusion, the automated fluorometric enzyme immunoassay tested in the present study produces reliable clinical results with a rapid turnaround time and therefore can be recommended for use in the early detection of AMI in a laboratory of Coronary Care Unit.

Coliforms and Enterobacteriaceae Isolates from Selected Foods.

We obtained coliform counts and Enterobacteriaceae counts using violet red bile agar (VRB) and VRB + 1% glucose (VRBG), respectively, of samples of five food products. From each set of VRB and VRBG plates, 28 to 40 "typical" colonies were randomly selected and identified by use of the R-B Enteric Differential System. A pure culture of each isolate was also subjected to the sequential tests for gas production in LST and BGLB broths (confirmed coliforms) and in EC broth at 45.5 C (fecal coliforms). IMViC reaction patterns of EC-positive cultures were also determined. Approximately 80% of the VRB isolates from broiler skin and from mechanically deboned poultry meat (MDPM) met all the criteria for fecal coliforms, whereas only 62.5% and 36.5%, respectively, of the VRBG isolates from these two products met these criteria. Fewer than 10% of the VRB and VRBG isolates from chicken pot pie, ground beef, or pork sausage produced gas in LST broth. The percentages of fecal coliforms and Escherichia coli (Type I or II) among the 179 VRB isolates were 34.1 and 33.5, respectively. Corresponding percentages for the 193 VRBG isolates were 20.7 and 19.7. E. coli was the predominant species isolated on both media from broiler skin and MDPM. Enterobacter agglomerans was the principal species isolated from chicken pot pie and pork sausage; Serratia liquefaciens predominated in ground beef.