Advancing multiproduct resin reuse for development and clinical manufacturing of an antibody-based therapeutic.

Chromatography resins used for purifying biopharmaceuticals are generally dedicated to a single product. For clinical manufacturing, this can result in resin being used only for a fraction of its potential lifetime. Extending the use of resins to multiple products can significantly reduce resin waste and cost. It can also improve manufacturing flexibility in case of raw material shortage during times such as the COVID-19 pandemic. The work presented herein describes an overarching multiproduct resin reuse (MRR) strategy, which includes a risk assessment, strategic planning, small-scale feasibility runs, and the successful execution of the MRR strategy to support Good manufacturing practice (GMP) clinical manufacturing of an antibody-based therapeutic. Specifically, an anion exchange (AEX) and cation exchange (CEX) MRR strategy is described. Clearance of carryover biological product is demonstrated by first cleaning the AEX and CEX manufacturing columns with sodium hydroxide to ensure inactivation and degradation of the carryover protein and followed by a blank buffer elution that is tested using various analytical methodologies to ensure reduction of the carryover protein to an acceptable level. To our knowledge, this is the first time an MRR approach has been successfully implemented and submitted to health authorities to support biologic GMP clinical manufacture.

Pyisotopomer: A Python package for obtaining intramolecular isotope ratio differences from mass spectrometric analysis of nitrous oxide isotopocules.

RATIONALE: Obtaining nitrous oxide isotopocule measurements with isotope ratio mass spectrometry (IRMS) involves analyzing the ion current ratios of the nitrous oxide parent ion (N2 O+ ) as well as those of the NO+ fragment ion. The data analysis requires correcting for "scrambling" in the ion source, whereby the NO+ fragment ion obtains the outer N atom from the N2 O molecule. While descriptions exist for this correction, and interlaboratory intercalibration efforts have been made, there has yet to be published a package of code for implementing isotopomer calibrations. METHODS: We developed a user-friendly Python package (pyisotopomer) to determine two coefficients (γ and κ) that describe scrambling in the IRMS ion source, and then used this calibration to obtain intramolecular isotope deltas in N2 O samples. RESULTS: With two appropriate reference materials, γ and κ can be determined robustly and accurately for a given IRMS system. An additional third reference material is needed to define the zero-point of the delta scale. We show that IRMS scrambling behavior can vary with time, necessitating regular calibrations. Finally, we present an intercalibration between two IRMS laboratories, using pyisotopomer to calculate γ and κ, and to obtain intramolecular N2 O isotope deltas in lake water unknowns. CONCLUSIONS: Given these considerations, we discuss how to use pyisotopomer to obtain high-quality N2 O isotopocule data from IRMS systems, including the use of appropriate reference materials and frequency of calibration.

Export of ice-nucleating particles from watersheds: results from the Amazon and Tocantins river plumes.

We examined ice-nucleating particles (INPs) in the plumes of the Tocantins and Amazon rivers, which drain watersheds with different proportions of degraded land. The concentration of INPs active at -15°C (INP-15) was an order of magnitude lower in the Tocantins (mean = 13.2 ml-1; s.d. = 7.8 ml-1), draining the more degraded watershed, compared with the Amazon (mean = 175.8 ml-1; s.d. = 11.2 ml-1), where the concentration was also significantly higher than in Atlantic surface waters (mean = 3.2 ml-1; s.d. = 2.3 ml-1). Differences in heat tolerance suggest that INPs emitted by the Amazon rainforest to the atmosphere or washed into the river might originate from contrasting sources on top of and below the rainforest canopy, respectively. For the Amazon River, we estimate a daily discharge of 1018 INP-15 to Atlantic waters. Rivers in cooler climate zones tend to have much higher concentrations of INPs and could, despite a smaller water volume discharged, transfer even larger absolute numbers of INP-15 to shelf waters than does the Amazon. To what extent these terrestrial INPs become aerosolized by breaking waves and bubble-bursting remains an open question.

[Observations on the influence of trauma-centered psychotherapy with EMDR therapy on somatic comorbidities using the example of Hashimoto's autoimmune thyroiditis]. / Einfluss der traumazentrierten Psychotherapie mit EMDR-Therapie auf somatische Komorbiditäten : Beobachtungen am Beispiel der Autoimmunthyreoiditis Hashimoto.

BACKGROUND: Results of modern research show a relationship between emotional stress and the occurrence of autoimmune diseases as a comorbidity. The authors use EMDR therapy (Eye Movement Desensitization and Reprocessing) to treat trauma disorders. They wondered whether and to what extent this treatment also affects autoimmune processes. METHOD: Parallel to the trauma-focused psychotherapy with EMDR, the thyroid hormone substitution dose was documented in patients with active Hashimoto's autoimmune thyroiditis requiring substitution. Hashimoto's autoimmune thyroiditis had already been diagnosed by a specialist and drug treatment had been initiated before starting outpatient psychotherapy. RESULTS AND CONCLUSION: So far in five cases a decrease in autoimmune activity and a stability of the results in the follow-up between six months and one year could be observed. It is now necessary to examine whether these results can be confirmed in a larger number of patients and a diversity of therapists and whether these observations can be transferred to other somatic comorbidities.

Unchanged nitrate and nitrite isotope fractionation during heterotrophic and Fe(II)-mixotrophic denitrification suggest a non-enzymatic link between denitrification and Fe(II) oxidation.

Natural-abundance measurements of nitrate and nitrite (NOx) isotope ratios (δ15N and δ18O) can be a valuable tool to study the biogeochemical fate of NOx species in the environment. A prerequisite for using NOx isotopes in this regard is an understanding of the mechanistic details of isotope fractionation (15ε, 18ε) associated with the biotic and abiotic NOx transformation processes involved (e.g., denitrification). However, possible impacts on isotope fractionation resulting from changing growth conditions during denitrification, different carbon substrates, or simply the presence of compounds that may be involved in NOx reduction as co-substrates [e.g., Fe(II)] remain uncertain. Here we investigated whether the type of organic substrate, i.e., short-chained organic acids, and the presence/absence of Fe(II) (mixotrophic vs. heterotrophic growth conditions) affect N and O isotope fractionation dynamics during nitrate (NO3 -) and nitrite (NO2 -) reduction in laboratory experiments with three strains of putative nitrate-dependent Fe(II)-oxidizing bacteria and one canonical denitrifier. Our results revealed that 15ε and 18ε values obtained for heterotrophic (15ε-NO3 -: 17.6 ± 2.8‰, 18ε-NO3 -:18.1 ± 2.5‰; 15ε-NO2 -: 14.4 ± 3.2‰) vs. mixotrophic (15ε-NO3 -: 20.2 ± 1.4‰, 18ε-NO3 -: 19.5 ± 1.5‰; 15ε-NO2 -: 16.1 ± 1.4‰) growth conditions are very similar and fall within the range previously reported for classical heterotrophic denitrification. Moreover, availability of different short-chain organic acids (succinate vs. acetate), while slightly affecting the NOx reduction dynamics, did not produce distinct differences in N and O isotope effects. N isotope fractionation in abiotic controls, although exhibiting fluctuating results, even expressed transient inverse isotope dynamics (15ε-NO2 -: -12.4 ± 1.3 ‰). These findings imply that neither the mechanisms ordaining cellular uptake of short-chain organic acids nor the presence of Fe(II) seem to systematically impact the overall N and O isotope effect during NOx reduction. The similar isotope effects detected during mixotrophic and heterotrophic NOx reduction, as well as the results obtained from the abiotic controls, may not only imply that the enzymatic control of NOx reduction in putative NDFeOx bacteria is decoupled from Fe(II) oxidation, but also that Fe(II) oxidation is indirectly driven by biologically (i.e., via organic compounds) or abiotically (catalysis via reactive surfaces) mediated processes co-occurring during heterotrophic denitrification.

Microbial niche differentiation explains nitrite oxidation in marine oxygen minimum zones.

Nitrite is a pivotal component of the marine nitrogen cycle. The fate of nitrite determines the loss or retention of fixed nitrogen, an essential nutrient for all organisms. Loss occurs via anaerobic nitrite reduction to gases during denitrification and anammox, while retention occurs via nitrite oxidation to nitrate. Nitrite oxidation is usually represented in biogeochemical models by one kinetic parameter and one oxygen threshold, below which nitrite oxidation is set to zero. Here we find that the responses of nitrite oxidation to nitrite and oxygen concentrations vary along a redox gradient in a Pacific Ocean oxygen minimum zone, indicating niche differentiation of nitrite-oxidizing assemblages. Notably, we observe the full inhibition of nitrite oxidation by oxygen addition and nitrite oxidation coupled with nitrogen loss in the absence of oxygen consumption in samples collected from anoxic waters. Nitrite-oxidizing bacteria, including novel clades with high relative abundance in anoxic depths, were also detected in the same samples. Mechanisms corresponding to niche differentiation of nitrite-oxidizing bacteria across the redox gradient are considered. Implementing these mechanisms in biogeochemical models has a significant effect on the estimated fixed nitrogen budget.

The response of the marine nitrogen cycle to ocean acidification.

Ocean acidification (OA), arising from the influx of anthropogenically generated carbon, poses a massive threat to the ocean ecosystems. Our knowledge of the effects of elevated anthropogenic CO2 in marine waters and its effect on the performance of single species, trophic interactions, and ecosystems is increasing rapidly. However, our understanding of the biogeochemical cycling of nutrients such as nitrogen is less advanced and lacks a comprehensive overview of how these processes may change under OA. We conducted a systematic review and meta-analysis of eight major nitrogen transformation processes incorporating 49 publications to synthesize current scientific understanding of the effect of OA on nitrogen cycling in the future ocean by 2100. The following points were identified by our meta-analysis: (a) Diazotrophic nitrogen fixation is likely enhanced by 29% ± 4% under OA; (b) species- and strain-specific responses of nitrogen fixers to OA were detectable, which may result in alterations in microbial community composition in the future ocean; (c) nitrification processes were reduced by a factor of 29% ± 10%; (d) declines in nitrification rates were not reflected by nitrifier abundance; and (e) contrasting results in unispecific culture experiments versus natural communities were apparent for nitrogen fixation and denitrification. The net effect of the nitrogen cycle process responses also suggests there may be a shift in the relative nitrogen pools, with excess ammonium originating from CO2 -fertilized diazotrophs. This regenerated inorganic nitrogen may recycle in the upper water column increasing the relative importance of the ammonium-fueled regenerated production. However, several feedback mechanisms with other chemical cycles, such as oxygen, and interaction with other climate change stressors may counteract these findings. Finally, our review highlights the shortcomings and gaps in current understanding of the potential changes in nitrogen cycling under future climate and emphasizes the need for further ecosystem studies.

Microbiological screenings for infection control in unaccompanied minor refugees: the German Armed Forces Medical Service's experience.

BACKGROUND: The German Military Medical Service contributed to the medical screening of unaccompanied minor refugees (UMRs) coming to Germany in 2014 and 2015. In this study, a broad range of diagnostic procedures was applied to identify microorganisms with clinical or public health significance. Previously, those tests had only been used to screen soldiers returning from tropical deployments. This instance is the first time the approach has been studied in a humanitarian context. METHODS: The offered screenings included blood cell counts, hepatitis B serology and microscopy of the stool to look for protozoa and worm eggs as well as PCR from stool samples targeting pathogenic bacteria, protozoa and helminths. If individuals refused certain assessments, their decision to do so was accepted. A total of 219 apparently healthy male UMRs coming from Afghanistan, Egypt, Somalia, Eritrea, Syria, Ghana, Guinea, Iran, Algeria, Iraq, Benin, Gambia, Libya, Morocco, Pakistan, and Palestine were assessed. All UMRs who were examined at the study department were included in the assessment. RESULTS: We detected decreasing frequencies of pathogens that included diarrhoea-associated bacteria [Campylobacter (C.) jejuni, enteropathogenic Escherichia (E.) coli (EPEC), enterotoxic E. coli (ETEC), enteroaggregative E. coli (EAEC), enteroinvasive E. coli (EIEC)/Shigella spp.), Giardia (G.) duodenalis, helminths (comprising Schistosoma spp., Hymenolepis (H.) nana, Strongyloides (S.) stercoralis] as well as hepatitis B virus. Pathogenic microorganisms dominated the samples by far. While G. duodenalis was detected in 11.4% of the assessed UMRs, the incidence of newly identified cases in the German population was 4.5 cases per 100,000 inhabitants. CONCLUSIONS: We conclude that the applied in-house PCR screening systems, which have proven to be useful for screening military returnees from tropical deployments, can also be used for health assessment of immigrants from the respective sites. Apparently healthy UMRs may be enterically colonized with a broad variety of pathogenic and apathogenic microorganisms. Increased colonization rates, as shown for G. duodenalis, can pose a hygiene problem in centralized homes for asylum seekers.

Beyond Ebola treatment units: severe infection temporary treatment units as an essential element of Ebola case management during an outbreak.

In the course of the Ebola outbreak in West Africa that was witnessed since early 2014, the response mechanisms showed deficits in terms of timeliness, volume and adequacy. The authors were deployed in the Ebola campaign in the West African country Liberia, where by September 2014 the changing epidemiological pattern made reconsiderations of guidelines and adopted procedures necessary. A temporary facility set up as a conventional Ebola Treatment Unit in the Liberian capital Monrovia was re-dedicated into a Severe Infections Temporary Treatment Unit. This facility allowed for stratification based on the nosocomial risk of exposure to Ebola virus for a growing subgroup of admitted patients that in the end would turn out as Ebola negative cases. At the same time, adequate diagnostic measures and treatment for the non-Ebola conditions of these patients could be provided without compromising work safety of the employed staff. The key elements of the new unit comprised a Suspect Cases Area similar to that of conventional Ebola treatment units for newly arriving patients, an Unlikely Cases Area for patients with a first negative Ebola PCR result, and a Confirmed Negative Cases Area for patients in whom Ebola could be ruled out. The authors, comprising representatives of the Liberian Ministry of Health and Social Welfare, as well as infectious disease specialists from the German Ebola Task Force are presenting key features of the adapted concept, and are highlighting its relevance in raising acceptance for outbreak counter-measures within the population at stake.

Low Enteric Colonization with Multidrug-Resistant Pathogens in Soldiers Returning from Deployments- Experience from the Years 2007-2015.

This assessment describes the enteric colonization of German soldiers 8-12 weeks after returning from mostly but not exclusively subtropical or tropical deployment sites with third-generation cephalosporin-resistant Enterobacteriaceae, vancomycin-resistant enterococci (VRE), and methicillin-resistant Staphylococcus aureus (MRSA). Between 2007 and 2015, 828 stool samples from returning soldiers were enriched in nonselective broth and incubated on selective agars for Enterobacteriaceae expressing extended-spectrum beta-lactamases (ESBL), VRE and MRSA. Identification and resistance testing of suspicious colonies was performed using MALDI-TOF-MS, VITEK-II and agar diffusion gradient testing (bioMérieux, Marcy-l'Étoile, France). Isolates with suspicion of ESBL were characterized by ESBL/ampC disc-(ABCD)-testing and molecular approaches (PCR, Sanger sequencing). Among the returnees, E. coli with resistance against third-generation cephalosporins (37 ESBL, 1 ESBL + ampC, 1 uncertain mechanism) were found in 39 instances (4.7%). Associated quinolone resistance was found in 46.2% of these isolates. Beta-lactamases of the blaCTX-M group 1 predominated among the ESBL mechanisms, followed by the blaCTX-M group 9, and blaSHV. VRE of vanA-type was isolated from one returnee (0.12%). MRSA was not isolated at all. There was no clear trend regarding the distribution of resistant isolates during the assessment period. Compared with colonization with resistant bacteria described in civilians returning from the tropics, the colonization in returned soldiers is surprisingly low and stable. This finding, together with high colonization rates found in previous screenings on deployment, suggests a loss of colonization during the 8- to 12-week period between returning from the deployments and assessment.

Surveillance of Food- and Smear-Transmitted Pathogens in European Soldiers with Diarrhea on Deployment in the Tropics: Experience from the European Union Training Mission (EUTM) Mali.

Introduction. Since 2013, European soldiers have been deployed on the European Union Training Mission (EUTM) in Mali. From the beginning, diarrhea has been among the most "urgent" concerns. Diarrhea surveillance based on deployable real-time PCR equipment was conducted between December 2013 and August 2014. Material and Methods. In total, 53 stool samples were obtained from 51 soldiers with acute diarrhea. Multiplex PCR panels comprised enteroinvasive bacteria, diarrhea-associated Escherichia coli (EPEC, ETEC, EAEC, and EIEC), enteropathogenic viruses, and protozoa. Noroviruses were characterized by sequencing. Cultural screening for Enterobacteriaceae with extended-spectrum beta-lactamases (ESBL) with subsequent repetitive sequence-based PCR (rep-PCR) typing was performed. Clinical information was assessed. Results. Positive PCR results for diarrhea-associated pathogens were detected in 43/53 samples, comprising EPEC (n = 21), ETEC (n = 19), EAEC (n = 15), Norovirus (n = 10), Shigella spp./EIEC (n = 6), Cryptosporidium parvum (n = 3), Giardia duodenalis (n = 2), Salmonella spp. (n = 1), Astrovirus (n = 1), Rotavirus (n = 1), and Sapovirus (n = 1). ESBL-positive Enterobacteriaceae were grown from 13 out of 48 samples. Simultaneous infections with several enteropathogenic agents were observed in 23 instances. Symptoms were mild to moderate. There were hints of autochthonous transmission. Conclusions. Multiplex real-time PCR proved to be suitable for diarrhea surveillance on deployment. Etiological attribution is challenging in cases of detection of multiple pathogens.

N and O isotope fractionation in nitrate during chemolithoautotrophic denitrification by Sulfurimonas gotlandica.

Chemolithoautotrophic denitrification is an important mechanism of nitrogen loss in the water column of euxinic basins, but its isotope fractionation factor is not known. Sulfurimonas gotlandica GD1(T), a recently isolated bacterial key player in Baltic Sea pelagic redoxcline processes, was used to determine the isotope fractionation of nitrogen and oxygen in nitrate during denitrification. Under anoxic conditions, nitrate reduction was accompanied by nitrogen and oxygen isotope fractionation of 23.8 ± 2.5‰ and 11.7 ± 1.1‰, respectively. The isotope effect for nitrogen was in the range determined for heterotrophic denitrification, with only the absence of stirring resulting in a significant decrease of the fractionation factor. The relative increase in δ(18)ONO3 to δ(15)NNO3 did not follow the 1:1 relationship characteristic of heterotrophic, marine denitrification. Instead, δ(18)ONO3 increased slower than δ(15)NNO3, with a conserved ratio of 0.5:1. This result suggests that the periplasmic nitrate reductase (Nap) of S. gotlandica strain GD1(T) fractionates the N and O in nitrate differently than the membrane-bound nitrate reductase (Nar), which is generally prevalent among heterotrophic denitrifiers and is considered as the dominant driver for the observed isotope fractionation. Hence in the Baltic Sea redoxcline, other, as yet-unidentified factors likely explain the low apparent fractionation.

Late stage infection in sleeping sickness.

At the turn of the 19(th) century, trypanosomes were identified as the causative agent of sleeping sickness and their presence within the cerebrospinal fluid of late stage sleeping sickness patients was described. However, no definitive proof of how the parasites reach the brain has been presented so far. Analyzing electron micrographs prepared from rodent brains more than 20 days after infection, we present here conclusive evidence that the parasites first enter the brain via the choroid plexus from where they penetrate the epithelial cell layer to reach the ventricular system. Adversely, no trypanosomes were observed within the parenchyma outside blood vessels. We also show that brain infection depends on the formation of long slender trypanosomes and that the cerebrospinal fluid as well as the stroma of the choroid plexus is a hostile environment for the survival of trypanosomes, which enter the pial space including the Virchow-Robin space via the subarachnoid space to escape degradation. Our data suggest that trypanosomes do not intend to colonize the brain but reside near or within the glia limitans, from where they can re-populate blood vessels and disrupt the sleep wake cycles.

Teaching the rectal examination with simulations: effects on knowledge acquisition and inhibition.

CONTEXT: Undergraduate medical curricula are often deficient in teaching physical examinations in intimate zones, such as the rectal examination. Student inhibition is assumed to substantially hamper both the acquisition of knowledge and the performance of these examinations in practice. OBJECTIVES: The two present studies examined the effects of low-fidelity (LFS) and high-fidelity (HFS) simulation on the acquisition of the necessary knowledge and inhibition about carrying out the rectal examination. In addition, we investigated the effects of the different sequencing of the two simulations (HFS-LFS versus LFS-HFS). METHODS: A manikin for the rectal examination was used to implement the LFS. Standardised patients (SPs) were used to implement the HFS. Study samples consisted of 41 (Study 1) and 188 (Study 2) female and male undergraduate medical students. Each student participated in two individual sessions of 30 minutes each. Half the students participated first in the HFS and then in the LFS and the other half participated in the simulations in the opposite order. Outcome measures were self-rated inhibition and knowledge tests. RESULTS: In both studies, HFS was found to reduce inhibition significantly more than LFS. Furthermore, in Study 2, a marginal effect of the sequence of simulation was found. In both studies, both types of simulation were found to facilitate the acquisition of knowledge. There was no sequence effect for the acquisition of knowledge. CONCLUSIONS: Teaching the rectal examination with the help of SPs, who represent an HFS, can help medical undergraduate students to overcome inhibition about this examination. Standardised patient simulation is far more effective than that achieved using a manikin, which represents an LFS. Both types of simulation support the acquisition of knowledge.

Marked differences in the prevalence of chloroquine resistance between urban and rural communities in Burkina Faso.

BACKGROUND: Chloroquine (CQ) resistance has reached high levels in Africa in recent years. Little is known about variations of resistance between urban and rural areas. OBJECTIVES: To compare the rates of in vivo resistance to CQ and the prevalences of the main molecular marker for CQ resistance among young children from urban and rural areas in Burkina Faso. METHODS: The current analysis used the frame of a randomized controlled trial (ISRCTN27290841) on the combination CQ-methylene blue (MB) (n=177) compared to CQ alone (n=45) in young children with uncomplicated malaria. We examined clinical and parasitological failure rates as well as the prevalence of the Plasmodium falciparum chloroquine resistance transporter gene (pfcrt) T76 mutation. RESULTS: Clinical and parasitological failure rates of CQ-MB differed significantly between urban (70%) and rural areas (29%, p<0.0001). Likewise, CQ failure rates were higher in the urban setting. Matching this pattern, pfcrt T76 was more frequently seen among parasite strains from urban areas (81%) when compared to rural ones (64%, p=0.01). In the presence of parasites exhibiting pfcrt T76, the odds of overall clinical failure were increased to 2.6-fold ([1.33, 5.16], p(LR)=0.005). CQ was detected at baseline in 21% and 2% of children from the urban and the rural study area, respectively (p(Chi)=0.002). CONCLUSION: Even within circumscribed geographical areas, CQ efficacy can vary dramatically. The differences in the prevalence of pfcrt T76 and in CQ failure rates are probably explained by a higher drug pressure in the urban area compared to the rural study area. This finding has important implications for national malaria policies.

Compliance of young children with ITN protection in rural Burkina Faso.

BACKGROUND: Insecticide-treated bed nets (ITNs) are known to be highly effective in reducing malaria morbidity and mortality. The effectiveness of ITNs is largely influenced by behavioural factors and not much is known regarding such factors under programme conditions. METHODS: This descriptive study was nested into a large ITN effectiveness study in rural Burkina Faso. During two cross-sectional surveys in the dry and rainy season of 2003, random samples of young children from nine representative villages (n = 180 per survey) were investigated for compliance with ITN protection and related behaviour. Data were collected through direct observations and through interviews with mothers. RESULTS: ITNs were perceived as very important for protection against mosquitoes and malaria particularly during the rainy season, but there were problems with their use during the dry season. Young children usually slept with their mother under the ITN and self-reported compliance was 66% and 98% during dry and rainy season, respectively (confirmed by direct observation in 34% and 79%, respectively). Important reasons for low compliance during the dry season were high temperatures inside houses and problems related to changing sleeping places during the night. CONCLUSION: Under programme conditions, compliance with ITN protection in young children is sufficient during the rainy season, but is rather low during the hot and dry season. Greater emphasis needs to be placed on information/education efforts to make people aware of the fact that the risk of contracting malaria may persist throughout the year.

Aging sensitizes toward ROS formation and lipid peroxidation in PS1M146L transgenic mice.

Mutations in the presenilins (PS) account for the majority of familial Alzheimer disease (FAD) cases. To test the hypothesis that oxidative stress can underlie the deleterious effects of presenilin mutations, we analyzed lipid peroxidation products (4-hydroxynonenal (HNE) and malondialdehyde) and antioxidant defenses in brain tissue and levels of reactive oxygen species (ROS) in splenic lymphocytes from transgenic mice bearing human PS1 with the M146L mutation (PS1M146L) compared to those from mice transgenic for wild-type human PS1 (PS1wt) and nontransgenic littermate control mice. In brain tissue, HNE levels were increased only in aged (19-22 months) PS1M146L transgenic animals compared to PS1wt mice and not in young (3-4 months) or middle-aged mice (13-15 months). Similarly, in splenic lymphocytes expressing the transgenic PS1 proteins, mitochondrial and cytosolic ROS levels were elevated to 142.1 and 120.5% relative to controls only in cells from aged PS1M146L animals. Additionally, brain tissue HNE levels were positively correlated with mitochondrial ROS levels in splenic lymphocytes, indicating that oxidative stress can be detected in different tissues of PS1 transgenic mice. Antioxidant defenses (activities of antioxidant enzymes Cu/Zn-SOD, GPx, or GR) or susceptibility to in vitro oxidative stimulation was unaltered. In summary, these results demonstrate that the PS1M146L mutation increases mitochondrial ROS formation and oxidative damage in aged mice. Hence, oxidative stress caused by the combined effects of aging and PS1 mutations may be causative for triggering neurodegenerative events in FAD patients.

Effects of insecticide-treated bednets during early infancy in an African area of intense malaria transmission: a randomized controlled trial.

OBJECTIVE: Insecticide-impregnated bednets and curtains have been shown by many studies to be effective against malaria. However, because of possible interactions with immunity development, treated bednets may cause no effect at all or even an increase in malaria morbidity and mortality in areas of high transmission. To clarify this issue, we did a randomized controlled trial to assess the long-term effects of bednet protection during early infancy. METHODS: A total of 3387 neonates from 41 villages in rural Burkina Faso were individually randomized to receive either bednet protection from birth (group A) or from age 6 months (group B). Primary outcomes were all-cause mortality in all study children and incidence of falciparum malaria in a representative subsample of the study population. FINDINGS: After a mean follow-up of 27 months, there were 129 deaths in group A and 128 deaths in group B rate ratio (RR) 1.0 (95% confidence interval (CI): 0.78-1.27)). Falciparum malaria incidence was lower in group A than in group B, during early (0-5 months) and late infancy (6-12 months) (RR 3.1, 95% CI: 2.0-4.9; RR 1.3, 95% CI: 1.1-1.6) and rates of moderate to severe anaemia were significantly lower during late infancy (11.5% vs 23.3%, P = 0.008), but there were no differences between groups in these parameters in children older than 12 months. CONCLUSION: The findings from this study provide additional evidence for the efficacy of insecticide-treated nets in young children living in areas of intense malaria transmission.