Systematic literature review assessing the overall costs and societal impacts of moderate-to-severe atopic dermatitis in Europe.

Atopic dermatitis (AD) is a chronic inflammatory disease, driven by type 2 inflammation. The condition manifests as moderate-to-severe disease in approximately 20% of adults with AD across Europe and is associated with a substantial burden on patients, society and healthcare systems. However, systematic assessments capturing the totality of disease burden associated with moderate-to-severe AD are limited; therefore, the overall impacts of the disease may be underestimated. A systematic literature review (SLR) was carried out to assess the overall costs of moderate-to-severe AD across Europe, including the financial, societal and humanistic impacts. PubMed, Embase and Cochrane databases were searched to identify relevant studies published between 1 January 2010 and 2 June 2020. Scientific conference proceedings, health technology assessment websites and patient association group websites were also searched for relevant information. Twenty-seven publications, corresponding to 22 unique studies, were included in the analysis. Total costs (direct and productivity losses) reached €20 695 per-person-per-year (PPPY) for adults with uncontrolled symptoms of moderate-to-severe AD. Direct medical costs ranged between €307 and €6993 PPPY; prescription medications and specialist dermatologist visits were the main contributors. Costs increased with disease severity or with uncontrolled disease. Patients with AD also incurred personal costs of €927 per year for healthcare items not reimbursed, which increased by 9% for those with moderate-to-severe forms. Annual work productivity losses comprised most of the total costs reported for adults with moderate-to-severe AD (up to 60.8% of the total burden) and were highest in those with uncontrolled disease (€13 702 PPPY). Patients with moderate-to-severe disease also experienced physical, emotional, and social impacts. The overall costs of moderate-to-severe AD greatly impact on healthcare systems, patients and society. Sustained control of moderate-to-severe AD, through effective treatment and care management, is essential to limit the burden caused by the disease.

An evaluation of the USEPA calculations of greenhouse gas emissions from anaerobic lagoons.

On 10 Apr. 2009, USEPA proposed and on 30 Oct. 2009 USEPA finalized reporting thresholds for a wide range of human-derived sources of greenhouse gas (GHG) as a first step in establishing emission limits in the United States. The only on-farm source category that required monitoring under the proposed and final rule was methane (CH(4)) and nitrous oxide (NO(2)) emissions from manure storage facilities. Our objective was to assess, through a literature review, the methodology used by USEPA to estimate current CH(4) emissions from uncovered anaerobic lagoons and the proposed methodology for reporting those emissions under the proposed rule. A review of the performance of uncovered anaerobic lagoons indicates that they are more effective at degrading volatile solids (VS) than predicted using parameters provided by USEPA that had been developed for anaerobic digesters. We also documented errors in the USEPA- and International Panel on Climate Change-estimated methane conversion factors for uncovered anaerobic lagoons. We suggest estimating CH(4) emissions from anaerobic lagoons based on VS degraded in the lagoon and B' (m(3) CH(4) generated kg(-1) VS destroyed). Our estimate of CH(4) released from uncovered anaerobic lagoons indicated the regulatory operation size threshold could be at least 65% smaller than predicted by USEPA in the proposed rule. Our calculated estimate of CH(4) emissions was substantially greater than the few estimates of CH(4) loss based on direct measurements on uncovered anaerobic lagoons. More research is needed before it will be possible to provide definitive estimates of CH(4) loss from uncovered anaerobic lagoons.

Viral epitopes and monoclonal antibodies: isolation of blocking antibodies that inhibit virus neutralization.

The inability of pathogenic animal viruses to be completely neutralized by antibodies can lead to chronic viral infections in which infectious virus persists even in the presence of excess neutralizing antibody. A mechanism that results in this nonneutralized fraction of virus was defined by the topographical relationships of viral epitopes identified with monoclonal antibodies wherein monoclonal antibodies bind to virus and sterically block the binding of neutralizing antibodies.

Catalytic hydrolysis of vasoactive intestinal peptide by human autoantibody.

Vasoactive intestinal peptide (VIP) labeled with 125I, [Tyr10-125I]VIP, can be hydrolyzed by immunoglobulin G (IgG) purified from a human subject, as judged by trichloroacetic acid precipitation and reversed-phase high-performance liquid chromatography (HPLC). The hydrolytic activity was precipitated by antibody to human IgG, it was bound by immobilized protein G and showed a molecular mass close to 150 kilodaltons by gel filtration chromatography, properties similar to those of authentic IgG. The Fab fragment, prepared from IgG by papain treatment, retained the VIP hydrolytic activity of the IgG. Peptide fragments produced by treatment of VIP with the antibody fraction were purified by reversed-phase HPLC and identified by fast atom bombardment-mass spectrometry and peptide sequencing. The scissile bond in VIP deduced from these experiments was Gln16-Met17. The antibody concentration (73.4 fmol per milligram of IgG) and the Kd (0.4 nM) were computed from analysis of VIP binding under conditions that did not result in peptide hydrolysis. Analysis of the antibody-mediated VIP hydrolysis at varying concentrations of substrate suggested conformity with Michaelis-Menton kinetics (Km). The values for Km (37.9 X 10(-9) M) and the turnover number kcat (15.6 min-1) suggested relatively tight VIP binding and a moderate catalytic efficiency of the antibody.

Phenotypic switching of antibiotic resistance circumvents permanent costs in Staphylococcus aureus.

Bacterial antibiotic resistance is often associated with a fitness cost in the absence of the antibiotic [1,2]. We have examined a resistance mechanism in Staphylococcus aureus that negates these costs. Exposure to gentamicin both in vitro and in vivo has been reported to result in the emergence of a gentamicin-resistant small colony variant (SCV)[3-8]. We show that the emergence of SCVs following exposure to gentamicin results from a rapid switch and that bacteria exposed to cycles of gentamicin followed by antibiotic-free medium repeatedly switched between a resistant SCV and a sensitive parental phenotype (revertants). The fitness of revertants relative to S. aureus with stable gentamicin resistance was greater in drug-free media, which suggests that S. aureus has evolved an inducible and reversible resistance mechanism that circumvents a permanent cost to fitness.

Environmentally constrained mutation and adaptive evolution in Salmonella.

The relationship between environment and mutation is complex [1]. Claims of Lamarkian mutation [2] have proved unfounded [3-5]; it is apparent, however, that the external environment can influence the generation of heritable variation, through either direct effects on DNA sequence [6] or DNA maintenance and copying mechanisms [7-10], or as a consequence of evolutionary processes [11-16]. The spectrum of mutational events subject to environmental influence is unknown [6] and precisely how environmental signals modulate mutation is unclear. Evidence from bacteria suggests that a transient recombination-dependent hypermutational state can be induced by starvation [5]. It is also apparent that changes in the mutability of specific loci can be influenced by alterations in DNA topology [10,17]. Here we describe a remarkable instance of adaptive evolution in Salmonella which is caused by a mutation that occurs in intermediate-strength osmotic environments. We show that the mutation is not 'directed' and describe its genetic basis. We also present compelling evidence in support of the hypothesis that the mutational event is constrained by signals transmitted from the external environment via changes in the activity of DNA gyrase.

Transplantable primate tumors induced by Rous sarcoma virus. I. Induction of tumors transplantable into young marmosets.

The tumors induced in white-lipped marmosets (Saguinus fuscicollis, S. nigricollis) by Rous sarcoma virus (RSV) of chicken origin (RSV-SR) were not transplantable to allogeneic hosts. In contrast, RSV rescued from these tumors (RSV-M) induced sarcomas that were transplantable to young but not to adult marmosets. The tumors induced by RSV-M and the transplants rapidly enlarged, metastasized to various organs, and killed the recipients 29-59 days post inoculation. Cell lines were readily established from all transplantable sarcomas. No virus expression was detected in transplantable tumor cell lines by electron microscopy or by biochemical and biological assays. However, RSV of the same subgroup as RSV-SR was rescued from both short-term and long-term tumor cell cultures by cocultivation with chicken embryo fibroblasts (CEF). The rescued viruses transformed marmoset cells 100-fold more efficiently than CEF cells, although CEF cells remained permissive for virus replication. Cytogenetic studies revealed extensive chromosome abnormalities in tumor transplants but not in RSV-M-induced sarcomas. All cell lines were hyperploid and contained structurally abnormal, large metacentric and telocentric chromosomes. Immunologic studies failed to detect group-specific (gs) antigen of the avian sarcoma-leukemia complex in either RSV-M-induced, transformed cells or tumor transplants. By complement-dependent cytotoxicity assays, with the use of marmoset anti-gs serum, RSV-associated antigen could be detected on the surfaces of tumor cells. No differences in the expression of this antigen existed between transplantable and nontransplantable marmoset sarcomas. All transplantable cell lines contained abnormal amounts of lipids and glycogen in comparison to RSV-SR-induced tumors and normal marmoset cell lines. The glycogen was associated with unique cytoplasmic membrane complexes and was surrounded by either single- or double-membraned vesicles.

Spontaneous human mononuclear cell cytotoxicity to cultured tumor cells: reproducibility of serial measurements with the use of a chromium-51-release microcytotoxicity assay.

Twenty-three healthy human adults with naturally occurring (spontaneous) mononuclear cell cytotoxicity (MCC) to cultured human tumor cells were evaluated in a serial study over 23 months to determine whether their initial MCC levels remained stable. A 3-hour 51Cr-release microcytotoxicity assay, in which the HBT-3 cancer-derived cell line with HeLa markers and the ALAB breast cancer-derived cell line were used, was standardized and its suitability for such serial studies was evaluated. The data showed that the 51Cr-release assay is reproducible and suitable for use in serial studies of MCC. The intrinsic technical error for a single assay averaged 12.5% (2-26%) coefficient of variation. The greatest sources of technical variation with time were the passage level of the target cells and the temperature at which the effector cells were stored and handled. When these technical factors were controlled, naturally occurring MCC was fairly reproducible with time. Of the healthy individuals with initial high or low MCC levels, 61% tended to retain these levels in a stable fashion, but the remaining 39% studied showed fluctuations of MCC greater than 33% coefficient of variation.

Immunization of Rous sarcoma virus-inoculated marmosets with BCG and transformed allogeneic cells.

The effects of specific immunotherapy with allogeneic cells transformed by Schmidt-Ruppin Rous sarcoma virus (SR-RSV), of treatment with BCG, and of surgery on the growth of SR-RSV-induced sarcomas in white-lipped marmosets were studied. Tumor incidence, tumor progression, and survival did not differ between control and treated animals. Animals immunized with BCG developed lymphocyte reactivity to tuberculin, which remained until the animals died. BCG was isolated from the spleen of one tumor-bearing animal.

Neuronal and synaptic measurements in the visual cortex of adult rats after undernutrition during normal or artificial rearing.

It is possible that the reported effects of early life undernutrition on brain morphology may be due to alterations in mother-infant interactions and not directly to undernutrition. We have investigated this possibility by comparing artificially reared with mother-reared rats. Four groups of black-and-white hooded male rats were reared. These consisted of mother reared control (MRC), mother reared undernourished (MRU), artificially reared control (ARC) and artificially reared undernourished (ARU). Artificially reared rats were raised in isolation away from their mothers from 5 to 21 days of postnatal age. They were fitted with a gastric cannula through which 'milk' was infused automatically. The period of undernutrition lasted from 5 to 25 postnatal days, following which the animals were fed ad libitum until 312 days of age. Rats from each group were then killed by perfusion with buffered 2.5% glutaraldehyde. Pieces of visual cortex from each rat were postfixed in osmium tetroxide and embedded in resin. Stereological procedures at the light and electron microscopical levels were used to estimate the synapse-to-neuron ratios in cortical layers II to IV. Both MRC and ARC rats had about 7000 synapses per neuron. However, this ratio was about 8300 in MRU rats whilst it was only about 5000 in ARU animals. The rearing x nutrition interaction was statistically significant at the 0.1% level. These changes in the synapse-to-neuron ratio were mainly due to alterations in the numerical densities of the synapses rather than that of neurons. These results demonstrate that environmental isolation, as a result of artificial rearing procedures, and concurrent undernutrition during the first three weeks of postnatal life, interact with one another to produce marked morphological changes in the adult rat brain. However, environmental isolation was not, by itself, sufficient to cause permanent changes in interneuronal connectivity.

A phenomenon: left-biassed asymmetrical eye-opening in artificially reared rat pups.

From 4 or 5 postnatal days, rat pups were either reared by their mothers (MR) or artificially reared (AR) without their mothers by infusion of 'milk' through a gastric cannula. In AR, but not MR, pups the left eye was more likely to open first than the right eye (P less than 0.001). The left-biassed eye-opening of AR pups may be a manifestation of stress-induced lateralisation of their brains.

Evidence of non-maternally mediated acceleration of eye-opening in 'enriched' artificially reared rat pups.

Rat pups were artificially reared, without their mothers, from postnatal day 5. They were exposed thereafter to impoverished or enriched environmental conditions. The enrichment included gentling, social interaction and homing experience. Enriched rats showed accelerated eye-opening compared with their impoverished littermates (P less than 0.02). This is the first demonstration of stimulation-induced acceleration of development which cannot have been maternally mediated.

Anxiety and anger among abusers of different substances.

The levels of trait anxiety and trait anger were compared for adult clients in residential treatment with primary diagnoses of alcoholism, cannabis dependence, cocaine abuse and opioid dependence. With a sample of 809 clients results suggested each of these groups to have significantly elevated anxiety and anger scores, compared to non-client samples. However, there were no significant differences on either anxiety or anger scores based on drug of choice.

Effects of early life undernutrition in artificially-reared rats. 2. Subsequent behaviour.

Four groups of rat pups were reared: mother-reared control (well-fed) and undernourished (MRC, MRU) and artificially-reared control and undernourished (ARC, ARU). AR rats were fed expressed rats' milk (days 5-7), mixtures of rats' milk and milk substitute (days 8-16), and milk substitute only (days 17-20) by intermittent infusion via a gastric cannula. Undernutrition, imposed from 5-25 days, was effected initially by underfeeding the mother (MRU) or by infusing restricted quantities of milk (ARU). Weaning was at 21 days and undernutrition from 21-25 days was by restricting the supply of solid food. All rats were fed ad lib from 25 days. From 18 weeks, male rats were subjected to 3 behavioural tests: open field, social behaviour (encounters between rats of like treatment group) and bar-pressing for food (CRF, FR5, VI 1', VI 2' and Extinction). Previously undernourished rats responded at higher rates on FR5, received fewer rewards on VI 2' and displayed more aggression during social interaction. These findings may reflect direct effects of nutrition on brain growth. In the open field and social situations ARU were generally less active than ARC animals, whereas MRU and MRC rats differed little. These results are discussed in terms of MRU pups being maternally "buffered" against the worst effects of undernutrition and ARC rats displaying characteristics of "early stimulated" animals.

Effects of early life undernutrition in artificially-reared rats: 3. Further studies of growth and behaviour.

Four groups of rat pups were reared: mother-reared control (well-fed) and undernourished (MRC, MRU) and artificially-reared control and undernourished (ARC, ARU). AR rats were fed expressed rats' milk (days 5-7), mixtures of rats milk and milk substitute (days 8-16), and milk substitute only (days 17-20) by intermittent infusion via a gastric cannula. Undernutrition, imposed from 5-25 days, was effected initially by underfeeding the mother (MRU) or by infusing restricted quantities of milk (ARU). Weaning was at 21 days and undernutrition from 21-25 days was by restricting the supply of solid food. All rats were fed ad lib from 25 days. From 18 weeks, male rats were subjected to 3 tests of behaviour: spontaneous alternation, conditioned food preference, and visual discrimination (Lashley stand). There were negative effects of both undernutrition and artificial rearing on body growth in weight and length. For two aspects of behaviour, preference for carbohydrate relative to protein (greater in U rats) and latency to jump (longer in U rats) the effects of early undernutrition were the same in both MR and AR animals. Hence, these effects cannot have been mediated through alterations in maternal care.

Ovarian Sertoli-Leydig cell tumor with retiform pattern.

Sertoli-Leydig cell tumors (SLCT) are sex cord-stromal tumors which exhibit testicular differentiation.1 They account for less than 0.2% of ovarian neoplasms in total but comprise 4% of ovarian tumors in females under 20 yrs of age.2 The morphological appearance of these tumors varies more widely than that of any other ovarian tumor except for the teratomas. Histologically they are now classified into 5 categories: well-differentiated, intermediately differentiated, poorly differentiated, heterologous and retiform.1 We describe the case of an ovarian Sertoli-Leydig cell tumor with retiform pattern in a 7 yr old girl.

The mutagenic properties of N-nitrosopeptides in the Ames test.

N-(N-Acetyl-L-prolyl)-N-nitrosoglycine (APNG) and N-(N-acetylvalyl)-N-nitrosoglycine (AVNG) are shown to exert mutagenic activity in the Salmonella/mammalian microsome mutagenicity (Ames) test. Positive responses are apparent for base-pair substitution mutation-detecting strains (TA1535, TA100 and TA102) both with and without the addition of S9-mix. It is concluded that both APNG and AVNG are direct-acting mutagens.

An investigation of the endogenous formation of apparent total N-nitroso compounds in conventional microflora and germ-free rats.

The endogenous formation of apparent total N-nitroso compounds (ATNC) has been investigated in germ-free (GF) and conventional (CV) microflora rats as a function of the drinking-water nitrate concentration. ATNC levels were below the 40 micrograms (N-NO)/kg detection limit in the blood, liver, kidney, spleen and small intestine of all CV and GF rats. For the CV rats ATNC were detected in concentrations of up to 370 micrograms (N-NO)/kg in the large intestine and up to 50 micrograms (N-NO)/kg in the stomach and there was a significant positive correlation between ATNC formation and the drinking-water nitrate level. Comparison of these results with those from GF rats showed that the ATNC in the stomach and large intestine of the CV animals were formed by microbial action, most probably involving bacterial nitrate-reductase activity.

N-nitroso compounds and polycyclic aromatic hydrocarbons in Icelandic smoked cured mutton.

Five samples of Icelandic smoked cured mutton were analysed for volatile N-nitrosamines, N- nitrosoamino acids, total N-nitroso content and several polycyclic aromatic hydrocarbons. Levels varied considerably from sample to sample but generally the levels of polycyclic aromatic hydrocarbons and N-nitrosamines (as judged by N- nitrosoproline , N- nitrosohydroxyproline and total N-nitroso content) varied in the same way. It is suggested that in this product the smoking procedure is responsible for both groups of contaminants and that the curing process is relatively unimportant.

Analysis of polycyclic aromatic hydrocarbons in UK total diets.

Analysis of UK total-diet samples for polycyclic aromatic hydrocarbons was carried out using a simplified sample clean-up and a high-performance liquid chromatography dual fluorescence detector system. The results indicate that cereals and oils/fats contribute the major part (approximately one third each) of the polycyclic aromatic hydrocarbons in these total diets. Fruit, sugars and vegetables provide much of the remainder (approximately one quarter) while meat, fish, milk and beverages make relatively minor contributions. These results are compared with others in the current literature on polycyclic aromatic hydrocarbons in foods. The levels in the UK diet seem to be at least as low as those found elsewhere.