High-throughput organ-on-chip platform with integrated programmable fluid flow and real-time sensing for complex tissue models in drug development workflows.

Drug development suffers from a lack of predictive and human-relevant in vitro models. Organ-on-chip (OOC) technology provides advanced culture capabilities to generate physiologically appropriate, human-based tissue in vitro, therefore providing a route to a predictive in vitro model. However, OOC technologies are often created at the expense of throughput, industry-standard form factors, and compatibility with state-of-the-art data collection tools. Here we present an OOC platform with advanced culture capabilities supporting a variety of human tissue models including liver, vascular, gastrointestinal, and kidney. The platform has 96 devices per industry standard plate and compatibility with contemporary high-throughput data collection tools. Specifically, we demonstrate programmable flow control over two physiologically relevant flow regimes: perfusion flow that enhances hepatic tissue function and high-shear stress flow that aligns endothelial monolayers. In addition, we integrate electrical sensors, demonstrating quantification of barrier function of primary gut colon tissue in real-time. We utilize optical access to the tissues to directly quantify renal active transport and oxygen consumption via integrated oxygen sensors. Finally, we leverage the compatibility and throughput of the platform to screen all 96 devices using high content screening (HCS) and evaluate gene expression using RNA sequencing (RNA-seq). By combining these capabilities in one platform, physiologically-relevant tissues can be generated and measured, accelerating optimization of an in vitro model, and ultimately increasing predictive accuracy of in vitro drug screening.

Application of optimization algorithms to adaptive motion control for repetitive process.

The application of optimization algorithms to adaptive motion control is proposed in this paper. In order to provide optimal system response, optimization algorithm is used as adjustment mechanism of controller coefficients. Most of optimization algorithms are not able to work in continuous optimization mode and with non-constant search space (i.e. dataset). For this reason, the introduction of a novel approach, Adaptive Procedure for Optimization Algorithms (APOA), that allows to apply most of optimization algorithms to adaptation process seems to be necessary. Such an algorithm is innovative, due to its universality in terms of applied optimization algorithm. Its application allows to obtain optimal motion control in modern electric drives. The proposed APOA is presented together with the novel desired-response adaptive system (DRAS) approach for repetitive processes. This solution provides unchanged system response regardless of plant parameters variation or external disturbances. The developed adaptive approach based on optimization algorithm is implemented in permanent magnet synchronous motor (PMSM) drive to adapt state feedback speed controller during moment of inertia variations. Since the proposed DRAS is model-free approach, the adaptation procedure is immune to issues related to plant parameters mismatch. The obtained results prove that proposed adaptive speed controller for PMSM assures desired system response regardless of the moment of inertia variation.

Repeated co-treatment with mirtazapine and aripiprazole reversed the schizophrenia-like behaviors and increased the brain-derived neurotrophic factor mRNA expression in the adult Sprague-Dawley rats exposed to glutathione deficit during early postnatal brain development.

Recent studies suggest that impaired glutathione synthesis and dopaminergic transmission are important factors in the pathophysiology of schizophrenia. Moreover, some studies have suggested that antidepressants are able to increase the activity of atypical antipsychotics which may efficiently improve the treatment of negative and some cognitive symptoms of schizophrenia. In the present study, we investigated the influence of repeated co-treated with mirtazapine and aripiprazole on the schizophrenia-like behavior and brain-derived neurotrophic factor (BDNF) mRNA expression in adult rats exposed to glutathione deficit during early postnatal development. Between the postnatal days p5-p16, male pups were treated with the inhibitor of glutathione synthesis, BSO (L-buthionine-(S,R)-sulfoximine) and the dopamine uptake inhibitor, GBR 12909 alone or in combination. Mirtazapine and aripiprazole were given repeatedly, once daily for 21 days before the tests. The behavioral and biochemical tests were performed in p90-92 rats. BSO given alone and in combination with GBR 12909 induced deficits in the studied behavioral tests and decreased the expression of BDNF mRNA. Repeated aripiprazole administration at a higher dose reversed these behavioral deficits. Co-treatment with an ineffective dose of aripiprazole and mirtazapine also abolished the behavioral deficits and biochemical changes, especially in the hippocampus in these rats. The present study indicated that the inhibition of glutathione synthesis in early postnatal development induced long-term deficits corresponding to schizophrenia-like behavior and decreased the BDNF mRNA expression in adult rats, and these behavioural and biochemical deficits were reversed by repeated treatment with a higher dose of aripiprazole and also by co-treatment with an ineffective dose of aripiprazole and mirtazapine. The above data suggest that this neurodevelopment rat model of schizophrenia-induced by glutathione deficit evoked by repeated treatment with BSO alone and together with GBR 12909 in early postnatal life may be useful for studies on the pathomechanism of schizophrenia.

Flexibility of little auks foraging in various oceanographic features in a changing Arctic.

Using GPS-tracked individuals, we compared foraging ecology and reproductive output of a High-Arctic zooplanktivorous seabird, the little auk Alle alle, between three years differing in environmental conditions (sea surface temperature). Despite contrasting environmental conditions, average foraging fights distance and duration were generally similar in all studied years. Also, in all years foraging locations visited by the little auk parents during short trips (ST, for chick provisioning) were significantly closer to the colony compared to those visited during long trips (LTs, mainly for adults' self-maintenance). Nevertheless, we also found some differences in the little auk foraging behaviour: duration of LTs was the longest in the coldest year suggesting more time for resting for adults compared to warmer years. Besides, birds foraged closer to the colony and in significantly colder water in the coldest year. Interestingly, these differences did not affect chick diet: in all the years, the energy content of food loads was similar, with the Arctic copepod, Calanus glacialis copepodite stage V being the most preferred prey item (>73% of items by number and >67% by energy content). Also chick survival was similar in all the study years. However, when examining chicks growth rate we found that their peak body mass was lower in warmer years suggesting that overall conditions in the two warm years were less favourable. While our results, demonstrate a great foraging flexibility by little auks, they also point out their vulnerability to changing environmental conditions.

Conjugate of Enkephalin and Temporin Peptides as a Novel Therapeutic Agent for Sepsis.

Antimicrobial peptides (AMPs) exhibit a wide spectrum of actions, ranging from a direct bactericidal effect to multifunctional activities as immune effector molecules. The aim of this study was to examine the anti-inflammatory properties of a DAL-PEG-DK5 conjugate composed of a lysine-rich derivative of amphibian temporin-1CEb (DK5) and dalargin (DAL), the synthetic Leu-enkephalin analogue. Detailed study of the endotoxin-neutralizing activity of the peptide revealed that DAL-PEG-DK5 interacts with LPS and the LPS binding protein (LBP). Moreover, DAL-PEG-DK5 prevented dimerization of TLR4 at the macrophage surface upon LPS stimulation. This inhibited activation of the NF-κB signaling pathway and markedly reduced pro-inflammatory cytokine production. Finally, we showed that aggregation of DAL-PEG-DK5 into amyloid-like structures induced by LPS neutralized the endotoxin proinflammatory activity. Consequently, DAL-PEG-DK5 reduced morbidity and mortality in vivo, in a mouse model of endotoxin-induced septic shock. Collectively, the data suggest that DAL-PEG-DK5 is a promising therapeutic compound for sepsis.

Flexibility of foraging strategies of the great skua Stercorarius skua breeding in the largest colony in the Barents Sea region.

BACKGROUND: Foraging strategies of seabird species often vary considerably between and within individuals. This variability is influenced by a multitude of factors including age, sex, stage of annual life cycle, reproductive status, individual specialization and environmental conditions. RESULTS: Using GPS-loggers, we investigated factors affecting foraging flight characteristics (total duration, maximal range, total distance covered) of great skuas Stercorarius skua of known sex breeding on Bjørnøya, Svalbard, the largest colony in the Barents Sea region. We examined influence of sex (females are larger than males), phase of breeding (incubation, chick-rearing), reproductive status (breeders, failed breeders) and bird ID (they are known for individual foraging specialization). Our analyses revealed that only bird ID affected foraging flight characteristics significantly, indicating a high degree of plasticity regardless of sex, reproductive status or phase of breeding. We recognized three main groups of individuals: 1) those preying mainly on other seabirds in the breeding colonies (6%), 2) those foraging at sea (76%) and kleptoparasiting other seabirds and/or foraging on fish and/or offal discarded by fishing vessels, and 3) those alternating between preying on other seabirds in breeding colonies and foraging at sea (18%). Despite marked size sexual dimorphism, we found no apparent sex differences in flight characteristics. Birds after egg- or chick-loss and thus not constrained as central foragers did not modify their foraging flights. CONCLUSIONS: Great skuas breeding on Bjørnøya displayed a high degree of plasticity regardless of sex, reproductive status or phase of breeding. We recognized groups of individuals regularly preying in the seabird colonies, foraging at sea, and alternating between both strategies. This suggests foraging specialization of some individuals.

Habitat foraging niche of a High Arctic zooplanktivorous seabird in a changing environment.

Here, we model current and future distribution of a foraging Arctic endemic species, the little auk (Alle alle), a small zooplanktivorous Arctic seabird. We characterized environmental conditions [sea depth, sea surface temperature (SST), marginal sea ice zone (MIZ)] at foraging positions of GPS-tracked individuals from three breeding colonies in Svalbard: one located at the southern rim of the Arctic zone (hereafter 'boreo-Arctic') and two in the high-Arctic zone on Spitsbergen ('high-Arctic'). The birds from one 'high-Arctic' colony, influenced by cold Arctic water, foraged in the shallow shelf zone near the colony. The birds from remaining colonies foraged in a wider range of depths, in a higher SST zone ('boreo-Arctic') or in the productive but distant MIZ (second 'high-Arctic' colony). Given this flexible foraging behaviour, little auks may be temporarily resilient to moderate climate changes. However, our fuzzy logic models of future distribution under scenarios of 1 °C and 2 °C SST increase predict losses of suitable foraging habitat for the majority of little auk colonies studied. Over longer time scales negative consequences of global warming are inevitable. The actual response of little auks to future environmental conditions will depend on the range of their plasticity and pace of ecosystem changes.

To add or not to add probiotics to infant formulae? An updated systematic review.

We updated evidence on the effects of the administration of probiotic-supplemented infant formulae (IF) compared with unsupplemented IF. Five databases were searched up to September 2016 for randomised controlled trials. Twenty publications were identified, including five new RCTs. Supplementation of IF with Bifidobacterium lactis Bb12, either alone or with Streptococcus thermophilus, had no effect on growth, respiratory illness, antibiotic use, stool frequency or consistency. However, there was a significant reduction in the number of episodes of gastrointestinal infections (Bb12) and a lower frequency of colic or irritability (when both strains were used). Lactobacillus johnsonii La1 had no effect on growth, gastrointestinal infections, or respiratory illness episodes. There were no effects of supplementation of IF with Bifidobacterium longum BL999, alone or with Lactobacillus rhamnosus LPR. L. rhamnosus GG was associated with better growth; it had no effect on colic/crying, or irritability, and it was associated with greater indexes of loose stools and a higher defecation frequency. Lactobacillus reuteri ATCC 55730 had no effect on growth, colic, crying, irritability, respiratory illness, antibiotic use, stool frequency, or stool consistency; however, it reduced the number of episodes of diarrhoea. L. reuteri DSM 17938 had no effect on growth, night-time sleeping, or flatulence, but it reduced the number of spitting episodes. Lactobacillus salivarius CEC5713 had no effect on growth, colic, crying, or irritability; however, it resulted in a significant reduction in the rate of diarrhoea and the number of episodes of respiratory symptoms. In conclusion, the administration of probiotic-supplemented formulae to healthy infants does not raise safety concerns with regard to growth and adverse effects. Some beneficial clinical effects are possible; however, there is no existing robust evidence to recommend their routine use. The latter conclusion may reflect the small amount of data on a specific probiotic strain(s) and outcomes, rather than a genuine lack of an effect.

ESC expert statement on the effects on mood of the natural cycle and progestin-only contraceptives.

Hormonal fluctuations during the natural cycle, as well as progestins used for hormonal contraception, can exert effects on mood especially in vulnerable women. Negative effects of levonorgestrel-releasing intrauterine contraception on mood are rare.

Systematic review with meta-analysis: early infant feeding and coeliac disease--update 2015.

BACKGROUND: New evidence emerged on early feeding practices and the risk of coeliac disease. AIM: To systematically update evidence on these practices to find out whether there is a need to revise current recommendations. METHODS: MEDLINE, EMBASE and the Cochrane Library were searched from July 2012 (end of last search) to February 2015 for studies of any design that assessed the effect of gluten consumption and breastfeeding on the development of coeliac disease and/or coeliac disease-related autoimmunity. RESULTS: We identified 21 publications, including two, new, large, randomised controlled trials performed in high-risk infants. Exclusive or any breastfeeding, as well as breastfeeding at the time of gluten introduction, did not reduce the risk of developing coeliac disease during childhood. For infants at high risk of developing coeliac disease, gluten introduction at 4 months of age in very small amounts, or at 6 or 12 months of age, resulted in similar rates of coeliac disease diagnosis in early childhood. Later gluten introduction was associated with later development of coeliac specific autoimmunity and coeliac disease during childhood, but not total risk reduction. Observational studies indicate that consumption of a higher amount of gluten at weaning may increase the risk for coeliac disease development. CONCLUSIONS: Infant feeding practices (breastfeeding, time of gluten introduction) have no effect on the risk of developing coeliac disease during childhood (at least at specific timeframes evaluated in the included studies), necessitating an update of current European recommendations.

Children with autism spectrum disorders show abnormal conditioned response timing on delay, but not trace, eyeblink conditioning.

Children with autism spectrum disorder (ASD) and age-matched typically-developing (TD) peers were tested on two forms of eyeblink conditioning (EBC), a Pavlovian associative learning paradigm where subjects learn to execute an appropriately-timed eyeblink in response to a previously neutral conditioning stimulus (CS). One version of the task, trace EBC, interposes a stimulus-free interval between the presentation of the CS and the unconditioned stimulus (US), a puff of air to the eye which causes the subjects to blink. In delay EBC, the CS overlaps in time with the delivery of the US, usually with both stimuli terminating simultaneously. ASD children performed normally during trace EBC, exhibiting no differences from TD subjects with regard to the learning rate or the timing of the conditioned response. However, when subsequently tested on delay EBC, subjects with ASD displayed abnormally-timed conditioned eye blinks that began earlier and peaked sooner than those of TD subjects, consistent with previous findings. The results suggest an impaired ability of children with ASD to properly time conditioned eye blinks which appears to be specific to delay EBC. We suggest that this deficit may reflect a dysfunction of the cerebellar cortex in which increases in the intensity or duration of sensory input can temporarily disrupt the accuracy of motor timing over short temporal intervals.

Review article: the management of acute gastroenteritis in children.

BACKGROUND: In 2008, the European Society for Paediatric Gastroenterology, Hepatology and Nutrition (ESPGHAN) and the European Society of Paediatric Infectious Disease (ESPID) developed evidence-based guidelines for the management of acute gastroenteritis (AGE) in children in Europe. AIM: To summarise data published subsequently to the ESPGHAN/ESPID guidelines. METHODS: MEDLINE and The Cochrane Library were searched in August 2012 for randomised controlled trials (RCTs) or their meta-analyses published after 2008. RESULTS: Efforts to improve the taste and/or efficacy of oral rehydration solution (ORS) continue, and some interventions are promising. While standard (over 24 h) nasogastric rehydration is still being used, new evidence confirms that rapid (over 4 h) rehydration is also effective. For intravenous rehydration, new evidence is available regarding rapid or ultrarapid and large-volume vs. standard-volume rehydration; as the new evidence is not consistent, until more data are available, the administration of 20 mL/kg seems appropriate. Convincing evidence has accumulated showing that ondansetron reduces the risk for vomiting; however, a clearance on safety in children is needed. New evidence has reconfirmed that in Europe, where zinc deficiency is rare, there is no benefit from the use of zinc. New data, although mainly from outside of Europe, have reconfirmed that either smectite or racecadotril is an effective adjunctive therapy to oral rehydration. There is a clear effect of using certain probiotics, such as Lactobacillus GG or S. boulardii. CONCLUSIONS: The update of current ESPGHAN/ESPID recommendations is warranted.

Systematic review: early infant feeding and the prevention of coeliac disease.

BACKGROUND: PREVENTCD, Prevent Coeliac Disease, is an international project investigating the hypothesis of possible induction of tolerance to gluten in genetically predisposed children through introducing small quantities of gluten during the period of breastfeeding. AIM: To summarise current knowledge on the possible relationship between early feeding practices and the risk of coeliac disease (CD). METHODS: The Cochrane Library, MEDLINE, and EMBASE databases were searched in May 2011, and the search was updated in January 2012, and again in July 2012. RESULTS: Breastfeeding (BF) and CD: some studies show a protective effect of BF, while others show no effect. No studies have shown a long-term preventive effect. BF at the time of gluten introduction and CD: Results from a meta-analysis of five observational case-control studies suggest that BF at gluten introduction is associated with a lower risk of CD compared with formula feeding. It is unclear whether BF provides a permanent protection or only delays the onset of CD. Timing of gluten introduction: The data suggest that both early (≤4 months) and late (≥7 months) introduction of gluten may increase the risk of CD. Amount of gluten at weaning (and later) and CD: One incident case-referent study documented that the introduction of gluten in large amounts compared with small or medium amounts increased the risk of CD. CONCLUSIONS: In the absence of clear evidence, in order to decrease the risk of later coeliac disease, it is reasonable to avoid both early (<4 months) and late (≥7 months) introduction of gluten, and to introduce gluten while the infant is still being breastfed. Future studies may clarify the remaining uncertainties.

[Hormonal (levonorgestrel) emergency contraception--effectiveness and mechanism of action]. / Antykoncepcja dorazna z uyciem lewonorgestrelu--skutecznosc i mechanizm dzialania.

Periodic abstinence and coitus interruptus are the most popular methods of contraception in Poland. Recent studies have provided us with evidence that the so-called "menstrual calendar" may be much less effective than it was believed. In these circumstances, promotion and use of safe and truly effective contraceptives is very important for Polish women. Emergency contraception (EC) is a method which could be used even in cases when other contraception methods have failed. Mechanism of action of levonorgestrel used for EC and possible disturbances in the process of implantation of the blastocyst in the endometrium, remain the source of heated discussion among medical professionals. The latest publications provide us with evidence that the use of levonorgestrel in EC neither alters endometrial receptivity nor impedes implantation. Hormonal EC effectiveness is another hot topic of gynecological endocrinology and statistics. There is, however, no better, safer, and more ethically accepted method of preventing unwanted pregnancy for patients in need of postcoital contraception.

A reliability study of the new Back Strain Monitor based on clinical trials.

A new Back Strain Monitor (BSM) device has been developed in order to measure, record and analyze movements of the lower back. The purpose of this study was to examine the inter-tester and the intra-tester reliability of the movement measurements given by the BSM accelerometers, and compare it with the reliability of two other conventional measurement methods: the Double Inclinometer method (DI) and the Modified-Modified Schober (MMS) method. The clinical studies included 23 participants (16 males, 7 females) with no recent history of lower back pain, who wore the device during a combination of different anatomical movements (flexion, extension, left lateral flexion and right lateral flexion of the lumber spine). The tests were conducted by three therapists (testers). The reliability results for the BSM accelerometers clearly outperform the results obtained for the DI and the MMS methods. The inter-tester reliability gives the Intra-Class Correlation (ICC) value of 0.95 for the BSM flexion, 0.89 for the DI flexion and 0.74 for the MMS. The intra-tester reliability gives the ICC value of 0.99 for BSM flexion, 0.94 for DI flexion and 0.77 for the MMS. The BSM accelerometers were highly reliable in assessing back movements, measuring these movements with less error than the DI and MMS methods.

Different roles of TiR8/Sigirr on toll-like receptor signaling in intrarenal antigen-presenting cells and tubular epithelial cells.

Toll-like receptors (TLRs) exist on both myeloid and intrinsic renal cells contributing to the initiation of innate immunity during renal infection with uropathogenic Escherichia coli. Toll-interleukin 1 receptor (IL-1R) (TIR)8/SIGIRR is an orphan receptor of the TLR/IL-1R family, which suppresses TLR signaling of immune cells and is highly expressed in the kidney. Lack of TIR8/SIGIRR is associated with enhanced renal chemokine signaling upon exposure to lipopolysaccharide (LPS). This was because of TIR8/SIGIRR expression on resident intrarenal myeloid cells rather than tubular epithelial cells which express it on basolateral and luminal membranes. The lack of TIR8/SIGIRR does not enhance TLR/IL-1R signaling in tubular epithelial cells as was observed in monocytes. TIR8/SIGIRR is induced in monocytes treated with LPS or tumor necrosis factor and interferon-gamma in a dose-dependent manner but was downregulated in treated tubule epithelial cells. This cell type-specific regulation and function did not relate to mRNA splice variants but was associated with N- and O-glycosylation of the receptor in renal cells of myeloid and nonmyeloid origin. Our studies show that resident myeloid cells contribute to TLR-mediated antimicrobial immunity in the kidney and that this function is controlled by Tir8/sigirr. TIR8/SIGIRR does not suppress TLR signaling in tubular epithelial cells, which supports their role as sensors of microbial infection in the kidney.

Oral contraceptives use and weight gain in women with a Central European life-style.

OBJECTIVES: The aim of the study was to check and present data on the relation between combined oral contraceptives (COC) use and body weight in young women living in Poland. METHODS: Observational study on the group of young women on COC and the control group of young women who never used COC. Candidates for study (145 women) and control (218 women) groups were recruited in selected gynecological clinics in Poland during the period of 1-31 January 2002 (with the use of randomization). RESULTS: Both groups (study and control) were almost identical in age, anthropometric characteristics, number of pregnancies and deliveries. On the basis of this investigation it was found that there was no relation between use of COC and weight gain. Observed (in both groups) weight gain in young women was most probably related rather to time-passing (patients were getting older). The higher risk for overweight and obesity was found in the group of young women who had already had problems with overweight in their childhood. and in the group of women with high weight gain during the first pregnancy. CONCLUSION: COC use is not associated with weight gain in young women with a Central European life-style.

The effectiveness of a body mass reduction program in obese women in perimenopausal and menopausal age compared with the effectiveness of such a program in younger (18-44 years of age) women living in Poland.

OBJECTIVE: The aim of the work was: to compare the effectiveness of body mass reducing program in women of perimenopausal (and menopausal) age with the effectiveness of such a program when it was applied to women aged 18-44 years. METHODS: The paper deals with observation study of the group of obese patients recruited and treated in Bialystok's Clinical Center for Cardiology and Body Mass Reduction. The authors summarize 12 months of clinical observation (as it was initially planned) of the two groups of obese women, taking part in the special body mass reduction program; Group I - 81 women in perimenopausal and menopausal age, Group II - 107 women in 18-44 years of age. The proposed treatment contained diet, physical exercises, psychological support and pharmacotherapy. RESULTS: The positive reaction for the treatment of obesity was less visible in obese women in perimenopausal and menopausal age, than in the group of obese women in 18-44 years of age (change of BMI in the group of younger women was -3.44 kg/m2 vs. -2.65 kg/m2 in older women). As it was observed, the weaker reaction for the proposed treatment in the group of older women, was not related to lower BMR, than in the group of younger obese women. Provisional result shows that use of HRT (Hormonal Replace Therapy) may probably result in better response of body mass reduction in perimenopausal and menopausal women. CONCLUSION: The high dropout of the patients included into the study does not allow to formulate unequivocal conclusions but it seems that body mass reduction program for women in perimenopausal and menopausal age should concentrate on building the motivation of the patients, HRT may play some role in improvement of effectiveness of such program but this should be confirmed by further studies.

Swaziland's Traditional Birth Attendants Survey.

The Traditional Birth Attendants (TBAs) Survey in Swaziland was undertaken between March 27th 1996 and April 8th 1996. The objective of the survey was to generate reliable information regarding activities of TBAs in Swaziland. The survey was conducted in 25 Chiefdoms sampled out of a total of 206 Chiefdoms registered in Swaziland. The total number of sampled respondents in the 25 Chiefdoms was 721. From the survey, it is estimated that there were probably 3000 TBAs in the country, and in the majority of cases such TBAs would be a 51-year old woman who herself had delivered six children and had worked as a TBA for at least 10 years. Between 9,000 and 12,000 deliveries are estimated to take place out of health facilities. Of these many, nearly 43.5% are assisted by "TBAs"; 16.3% of woman interviewed deliver relative/family member and 15.1% are assisted by friends/neighbours, etc. Some of TBAs carry out procedures which are considered to be potentially harmful. Nearly 30% of TBAs have administered herbs; 45% attend to abnormal deliveries (breech and multiple pregnancies); 26.7% re-use their cord cutting tools and in the case of haemorrhage 23.4% do manual procedures within reproductive tract of delivering women.

Characterization of splenic glycosaminoglycans accumulated in vivo in experimentally induced amyloid-susceptible and amyloid-resistant mice.

The pathogenesis of the amyloid deposition diseases is poorly understood. The CE/J mouse, which is naturally protected from amyloid A (AA) protein amyloidosis, has provided a tool to study mechanisms that may be implicated in amyloid deposition diseases by means of comparison of findings with those in an AA-susceptible mouse strain. We have compared proteoglycan/glycosaminoglycan accumulation in vivo in amyloid-protected CE/J mouse strain and in AA-susceptible CBA/J mouse strain in homeostasis and when injected with amyloid-inducing agents. Results indicate that there is an overall increase in [(35)S]proteoglycan/glycosaminoglycan accumulation in the spleens of both strains of mice, but with a specific increase in heparan sulfate in only CBA/J mouse spleens that are rich in amyloid. Further, we report the absence of heparan sulfate in the splenic perifollicular areas of amyloid-free CE/J mouse, whereas in the amyloid-laden CBA/J mouse there is co-localization of heparan sulfate with the AA deposits. We have also examined the glycosaminoglycan disaccharide products in both these strains of mice for their sulfation positions and found no differences in the disaccharide composition of chondroitin/dermatan sulfate and heparan sulfate isolated from the control CBA/J and control CE/J mice. There were no differences in chondroitin/dermatan sulfate in both strains after experimental induction. However, analysis of the heparan sulfate disaccharides by means of capillary high-performance liquid chromatography linked to microelectrospray ionization time-of-flight mass spectrometry indicated that the disaccharide composition of the splenic heparan sulfate obtained from the treated CBA/J mice that had developed amyloid was markedly different from that obtained from the control CBA/J mice and the treated amyloid-resistant CE/J mice. These findings suggest that unique heparan sulfates play a fundamental role in the pathogenesis of amyloid.