Components Of Body Fluids Research Articles

Advanced forensic validation for human spermatozoa identification using SPERM HY-LITER™ Express with quantitative image analysis.

Identification of human semen is indispensable for the investigation of sexual assaults. Fluorescence staining methods using commercial kits, such as the series of SPERM HY-LITER™ kits, have been useful to detect human sperm via strong fluorescence. These kits have been examined from various forensic aspects. However, because of a lack of evaluation methods, these studies did not provide objective, or quantitative, descriptions of the results nor clear criteria for the decisions reached. In addition, the variety of validations was considerably limited. In this study, we conducted more advanced validations of SPERM HY-LITER™ Express using our established image analysis method. Use of this method enabled objective and specific identification of fluorescent sperm's spots and quantitative comparisons of the sperm detection performance under complex experimental conditions. For body fluid mixtures, we examined interference with the fluorescence staining from other body fluid components. Effects of sample decomposition were simulated in high humidity and high temperature conditions. Semen with quite low sperm concentrations, such as azoospermia and oligospermia samples, represented the most challenging cases in application of the kit. Finally, the tolerance of the kit against various acidic and basic environments was analyzed. The validations herein provide useful information for the practical applications of the SPERM HY-LITER™ Express kit, which were previously unobtainable. Moreover, the versatility of our image analysis method toward various complex cases was demonstrated.

In vitro model of production of antibodies; a new approach to reveal the presence of key bacteria in polymicrobial environments.

BackgroundThere is a rapid emergence of multiple resistant gram-negative bacteria due to overuse of antibiotics in the treatment of infections. Biofilms consist of polymicrobial communities that survive the host’s defense system. The key bacteria in biofilms are slow growing and support an attachment and rapid growth of other microorganisms. Current antimicrobial strategies often fail due to poor diagnosis of key pathogens in biofilms.The study aims to develop anti-bacterial human antibodies in vitro from patients who had recently undergone a systemic infection by pathogenic bacteria and to use these antibodies as a tool for detecting bacteria in biofilms.MethodsLymphocytes were separated from whole blood of patients (n = 10) and stimulated with heat-killed bacteria to produce antibodies in vitro. The specificity of antibodies in recognizing the bacteria against which they were directed was evaluated by surface plasmon resonance system (SPR) and electron microscopy. The ulcer secretions from patients with chronic and acute leg ulcers and healthy controls were analyzed by the SPR system and the results were compared with culture studies.ResultsThe produced antibodies recognized bacteria with high sensitivity (SPR). The antibodies against Enterococcus fecalis bound specifically to the microorganism in a bacterial co-culture that was visualized by electron microscopy.ConclusionIn the present work, a method for producing specific antibodies against bacteria is introduced to recognize bacterial components in body fluids of patients suffering from pathogenic biofilms. This diagnostic technique may be most useful in clinical microbiology and in the choice of antibiotics in the treatment of serious infections.Electronic supplementary materialThe online version of this article (doi:10.1186/s12866-016-0821-5) contains supplementary material, which is available to authorized users.

In Situ Evaluation of Calcium Phosphate Nucleation Kinetics and Pathways during Intra- and Extrafibrillar Mineralization of Collagen Matrices.

We revealed that nucleation sites within collagen fibrils determined pathways for calcium phosphate (CaP) nucleation and its transformation, from amorphous species to crystalline plates, during the biomineralization process. Using in situ small-angle X-ray scattering (SAXS), we examined the nucleation and growth of CaP within collagen matrices and elucidated how a nucleation inhibitor, polyaspartic acid (pAsp), governs mineralization kinetics and pathways at multiple length scales. Mineralization without pAsp led initially to spherical aggregates of CaP in the entire extrafibrillar spaces. With time, the spherical aggregates transformed into plates at the outermost surface of the collagen matrix, preventing intrafibrillar mineralization inside. However, mineralization with pAsp led directly to the formation of intrafibrillar CaP plates with a spatial distribution gradient through the depth of the matrix. The results illuminate mineral nucleation kinetics and real-time nanoparticle distributions within organic matrices in solutions containing body fluid components. Because the macroscale mechanical properties of collagen matrices depend on their mineral content, phase, and arrangement at the nanoscale, this study contributes to better design and fabrication of biomaterials for regenerative medicine.

Nanotechnology for the diagnosis and treatment of diseases.

Nanotechnology for the diagnosis and treatment of diseases.

7D.04

Objective: Our previous study demonstrated that one week of salt loading significantly impaired skin microvascular reactivity without changes in blood pressure (BP) levels. The aim of this study was to evaluate whether one week of high-salt (HS) diet changes body fluid status and body composition subsequently affecting vascular reactivity. Design and method: 10 healthy women (age range 20–23) took a 7-days low-salt (LS) diet (<40mmol Na/day) and 7-days HS diet (∼240mmol of Na/day). Salt resistance, defined as a <5 mmHg change in BP determined while on the LS and HS diets was confirmed in all subjects. Skin microvascular post occlusive reactive hyperemic (PORH) blood flow (indicator of endothelial function) was assessed by laser Doppler flowmetry (LDF) before and after each diet protocol. Plasma Renin Activity (PRA), plasma aldosterone, plasma and 24h-urine sodium, potassium, urea and creatinine levels were measured before and after diets. Body composition was assessed with a four-terminal portable impedance analyzer (Maltron Bioscan 920-II). Body Mass Index (BMI), Fat Free Mass% (FFM%), Fat Mass% (FM%), Total Body Water% (TBW%), Extracellular Water% (ECW%), Intracellular Water (ICW%), ECW/ICW, Plasma Fluid (PF), Interstitial Fluid (IF) and Body Density Mass (BDM) were calculated. Results: Changes in 24 h urinary sodium, PRA and plasma aldosterone levels confirmed subjects conformed to the diet. There was no change in BP and HR before and after both diet protocols. HS diet caused significant impairment in microvascular reactivity (PORH) (R-O LS diet 156 ± 23% vs. HS diet 100 ± 12%, P = 0.040). One week HS diet did not induce any significant change in body composition parameters BMI, FFM%, FM% and BDM, compared to LS diet. Body fluid components (TBW%, ECW%, ICW%, ECW/ICW, PF and IF) were not different in LS compared to HS group. Conclusions: This study confirmed that even one week of HS diet significantly altered microvascular reactivity in young healthy normotensive and salt-resistant women, without changes in BP. Furthermore, our results indicate that vascular changes after HS diet are independent of body composition and body fluid status just as they are pressure independent, but are consequence of unique effect of HS on endothelial function.

Plasma genetic and genomic abnormalities predict treatment response and clinical outcome in advanced prostate cancer

Liquid biopsies, examinations of tumor components in body fluids, have shown promise for predicting clinical outcomes. To evaluate tumor-associated genomic and genetic variations in plasma cell-free DNA (cfDNA) and their associations with treatment response and overall survival, we applied whole genome and targeted sequencing to examine the plasma cfDNAs derived from 20 patients with advanced prostate cancer. Sequencing-based genomic abnormality analysis revealed locus-specific gains or losses that were common in prostate cancer, such as 8q gains, AR amplifications, PTEN losses and TMPRSS2-ERG fusions. To estimate tumor burden in cfDNA, we developed a Plasma Genomic Abnormality (PGA) score by summing the most significant copy number variations. Cox regression analysis showed that PGA scores were significantly associated with overall survival (p < 0.04). After androgen deprivation therapy or chemotherapy, targeted sequencing showed significant mutational profile changes in genes involved in androgen biosynthesis, AR activation, DNA repair, and chemotherapy resistance. These changes may reflect the dynamic evolution of heterozygous tumor populations in response to these treatments. These results strongly support the feasibility of using non-invasive liquid biopsies as potential tools to study biological mechanisms underlying therapy-specific resistance and to predict disease progression in advanced prostate cancer.

Clinical relevance and contemporary methods for counting blood cells in body fluids suspected of inflammatory disease.

In many inflammatory diseases, the cellular components in body fluids [cerebrospinal fluid (CSF), serous fluids] are increased, rendering essential diagnostic information. The diagnostic value of the total white blood cell count (WBC) and differential count has been evaluated extensively over the years, and a remarkable amount of knowledge has been gained; yet, there is a great deal of clinical uncertainty whether the diagnosis should be based solely on these variables. In some diseases, such as peritonitis, the total WBC and differential count has high sensitivity; whereas, in differentiating pleural effusions, it lacks the sensitivity required to be clinically useful. Nevertheless, many guidelines consider these tests as cornerstone parameters, and in combination with clinical variables, they can successfully guide clinical decision making in initiating or postponing a treatment course for infection and/or inflammatory diseases while awaiting culture results. Although other methods are available for detecting and differentiating WBCs in body fluids, manual microscopy is still considered the gold standard despite its many limitations. During the last decade, automated analyzers have become a popular method for first line screening. Continued progress in their design has led to major improvements including their speed, improved accuracy and lower variability compared with microscopy. Disadvantages of this method include high imprecision in low ranges (depending on the method) and interfering factors. In a time where automation is at the front line in clinical laboratories, it is essential the results obtained are precise, accurate and reproducible. This review provides an overview of the relevance for cell counting in a variety of diagnostic body fluids, and highlights the current technologies used.

What Bathers Put into a Pool: A Critical Review of Body Fluids and a Body Fluid Analog

Bathers add inorganic and organic materials to pool water primarily via sweat and urine but also via sunscreens and cosmetics. The organic nitrogen (organic-N) compounds react with chlorine to form organic chloramines that have no biocidal effectiveness, volatile inorganic chloramines, and other oxidation products that create high combined chlorine measurements and chlorinous odors. Recent research on the reactions of free available chlorine (FAC) with organic-N components of body fluids and models of other organic-N compounds likely to be present in pool water is reviewed. One recipe for a body fluid analog (BFA) was found in model pool studies to reproduce a field-observed median chlorine demand of 5.5 g/standard bather and to contain a reduced nitrogen mass per average bather within the ranges of published estimates. Guidelines for the combined chlorine measurement are routinely exceeded in operating pools and in model pool studies even at modest bather loads simulated using the BFA recipe.

Polyvinyl alcohol-based hydrogel dressing gellable on-wound via a co-enzymatic reaction triggered by glucose in the wound exudate.

Glucose is a common component of body fluids. We describe a hydrogel wound dressing that can be obtained by pouring an aqueous solution of a polyvinyl alcohol derivative possessing phenolic hydroxyl moieties (PVA-Ph), containing glucose oxidase (GOx) and horseradish peroxidase (HRP), onto a wound. The in situ hydrogelation progresses through GOx-catalysed hydrogen peroxide (H2O2) generation from glucose in the wound exudate and HRP-catalysed cross-linking between phenolic hydroxyl moieties in PVA-Ph by consuming the H2O2. The potency as a wound dressing is demonstrated by experiments which reveal that the existence of the conditions inducing hydrogelation within 20 s at normal blood glucose concentration, the durability of the resultant hydrogels to compression and stretching, and the faster closure of full-thickness wounds created on rats treated with the in situ formed hydrogel than those treated by a commercially available hydrogel dressing.

Microfluidic sample preparation for diagnostic cytopathology.

The cellular components of body fluids are routinely analyzed to identify disease and treatment approaches. While significant focus has been placed on developing cell analysis technologies, tools to automate the preparation of cellular specimens have been more limited, especially for body fluids beyond blood. Preparation steps include separating, concentrating, and exposing cells to reagents. Sample preparation continues to be routinely performed off-chip by technicians, preventing cell-based point-of-care diagnostics, increasing the cost of tests, and reducing the consistency of the final analysis following multiple manually-performed steps. Here, we review the assortment of biofluids for which suspended cells are analyzed, along with their characteristics and diagnostic value. We present an overview of the conventional sample preparation processes for cytological diagnosis. We finally discuss the challenges and opportunities in developing microfluidic devices for the purpose of automating or miniaturizing these processes, with particular emphases on preparing large or small volume samples, working with samples of high cellularity, automating multi-step processes, and obtaining high purity subpopulations of cells. We hope to convey the importance of and help identify new research directions addressing the vast biological and clinical applications in preparing and analyzing the array of available biological fluids. Successfully addressing the challenges described in this review can lead to inexpensive systems to improve diagnostic accuracy while simultaneously reducing overall systemic healthcare costs.

Worsening of malnutrition-inflammation score is associated with high ratio of extracellular to total body water in hemodialysis patients

The Malnutrition-Inflammation Score (MIS) has been reported to be associated with mortality in hemodialysis (HD) patients. Bioelectrical impedance analysis is useful for assessing the body fluid components. The purpose of this study is to evaluate the association between change in MIS and fluid components in HD patients. A total of 76 patients who were receiving a 5-hour HD, 3 times weekly, were enrolled in this study. MIS was performed every 6 months. Using IN BODY 720, body fluid components were estimated once a year, including intracellular water (ICW), extracellular water (ECW), total body water (TBW), total body fat, body fat percentage and ECW/TBW ratio. In univariate analysis, worsening of MIS was correlated positively with age and ECW/TBW ratio. In multivariate analysis, worsening of MIS was significantly associated with higher ECW/TBW ratio. Higher ECW/TBW ratio remained significantly correlated with worsening of MIS in non-obese HD patients, but not in obese patients. In conclusion, non-obese HD patients who has higher ECW/TBW ratio should be considered to be a risk group for malnutrition.

Electromembrane extraction of amino acids from body fluids followed by capillary electrophoresis with capacitively coupled contactless conductivity detection

Electromembrane extraction (EME) proved to be a simple and rapid pretreatment method for analysis of amino acids and related compounds in body fluid samples. Body fluids were acidified to the final concentration of 2.5 M acetic acid and served as donor solutions. Amino acids, present as cations in the donor solutions, migrated through a supported liquid membrane (SLM) composed of 1-ethyl-2-nitrobenzene/bis-(2-ethylhexyl)phosphonic acid (85:15 (v/v)) into the lumen of a porous polypropylene hollow fiber (HF) on application of electric field. The HF was filled with 2.5 M acetic acid serving as the acceptor solution. Matrix components in body fluids were efficiently retained on the SLM and did not interfere with subsequent analysis. Capillary electrophoresis with capacitively coupled contactless conductivity detection was used for determination of 17 underivatized amino acids in background electrolyte solution consisting of 2.5 M acetic acid. Parameters of EME, such as composition of SLM, pH and composition of donor and acceptor solution, agitation speed, extraction voltage, and extraction time were studied in detail. At optimized conditions, repeatability of migration times and peak areas of 17 amino acids was better than 0.3% and 13%, respectively, calibration curves were linear in a range of two orders of magnitude ( r 2 = 0.9968–0.9993) and limits of detection ranged from 0.15 to 10 μM. Endogenous concentrations of 12 amino acids were determined in EME treated human serum, plasma, and whole blood. The method was also suitable for simple and rapid pretreatment and determination of elevated concentrations of selected amino acids, which are markers of severe inborn metabolic disorders.

Hydration and health: a review

Summary Water is essential for life and maintaining optimal levels of hydration is important for humans to function well. Water makes up a large proportion of our body weight (60% on average), distributed between the intracellular (inside cells) and extracellular (water in the blood and in between cells) compartments. Water is the major component of body fluids, such as blood, synovial fluid (fluid in the joints), saliva and urine, which perform vital functions in the body. The concentration of solutes (osmolality) in body fluids is closely controlled, and even very small changes in osmolality trigger a physiological response; either to increase body water by reducing urinary output and stimulating thirst; or to excrete excess water as urine. Generally, body water is maintained within narrow limits. However, if water losses are not sufficiently replaced, dehydration occurs. Extreme dehydration is very serious and can be fatal. More mild dehydration (about 2% loss of body weight) can result in headaches, fatigue and reduced physical and mental performance. It is also possible to consume too much water and in rare cases this can result in hyponatraemia (low levels of sodium in the blood). We can get water from almost all drinks and from some foods in the diet. Food provides about 20% on average and this could vary widely depending on the types of food chosen. We also get water from all the drinks we consume, with the exception of stronger alcoholic drinks like wines and spirits. All these can contribute to dietary water, but also have other effects on health both positive and negative. The major concerns with regards to beverages are their energy content and their effect on dental health. With obesity levels continuing to increase it is important for many in the population to control their energy intake, and drinks as well as foods must be considered for their energy content. With regards to dental health, there are two concerns; dental caries and dental erosion. Dental caries are caused by a reduction in pH due to bacterial fermentation of carbohydrates, and so the frequency of consumption of drinks containing sugars is a concern for risk of caries. Dental erosion occurs at a lower pH and is caused by the consumption of acidic foods and drinks, in particular, citrus juices and soft drinks containing acids. Individual water needs vary widely depending on many factors including body size and composition, the environment and levels of physical activity. Thus it is very difficult to make generic recommendations about the amount of water to consume. The FSA currently recommends drinking about 1.2 litres per day (about 6‐8 glasses).

Generation of an affinity matrix useful in the purification of natural inhibitors of plasmepsin II, an antimalarial‐drug target

Various approaches for removal of high-abundance components in body fluids are currently available. While most methods are constructed for plasma depletion, there is a need for body-fluid-specific strategies. The aim of the present study was to design an affinity matrix suitable for the depletion of high-abundance proteins in CSF (cerebrospinal fluid). Hence, molecules with specific affinity towards proteins present at high concentration in CSF were desired. Affibody molecules are specific binders of small size that have shown high stability under various conditions and are therefore good candidates for such a matrix. The protein composition in CSF resembles that in plasma. However, 20% of the proteins are brain-derived and are therefore present in higher proportions in CSF than in plasma, whereas larger plasma-derived proteins are less abundant in CSF. Therefore five high-abundance CSF proteins were chosen for the design of a CSF-specific depletion setup. Affibody molecules with specificity towards HSA (human serum albumin), IgG, transferrin and transthyretin were combined in an affinity column. In addition, polyclonal antibodies against cystatin C were coupled to chromatographic beads and packed in a separate column. Highly reproducible and efficient removal of the five target proteins was observed. The proportion of depleted proteins were estimated to be 99, 95, 74, 92 and 83% for HSA, IgG, transferrin, transthyretin and cystatin C respectively. SDS/PAGE analysis was used for monitoring and identifying proteins in native CSF, depleted CSF samples and the captured fractions. Moreover, shotgun proteomics was used for protein identification in native as well as depleted CSF and the achieved data were compared. Enhanced identification of lower-abundance components was observed in the depleted fraction, in terms of more detected peptides per protein.

Coagulation Factors IX and X Enhance Binding and Infection of Adenovirus Types 5 and 31 in Human Epithelial Cells

Most adenoviruses bind directly to the coxsackie and adenovirus receptor (CAR) on target cells in vitro, but recent research has shown that adenoviruses can also use soluble components in body fluids for indirect binding to target cells. These mechanisms have been identified upon addressing the questions of how to de- and retarget adenovirus-based vectors for human gene and cancer therapy, but the newly identified mechanisms also suggest that the role of body fluids and their components may also be of importance for natural, primary infections. Here we demonstrate that plasma, saliva, and tear fluid promote binding and infection of adenovirus type 5 (Ad5) in respiratory and ocular epithelial cells, which corresponds to the natural tropism of most adenoviruses, and that plasma promotes infection by Ad31. By using a set of binding and infection experiments, we also found that Ad5 and Ad31 require coagulation factors IX (FIX) or X (FX) or just FIX, respectively, for efficient binding and infection. The concentrations of these factors that were required for maximum binding were 1/100th of the physiological concentrations. Preincubation of virions with heparin or pretreatment of cells with heparinase I indicated that the role of cell surface heparan sulfate during FIX- and FX-mediated adenovirus binding and infection is mechanistically serotype specific. We conclude that the use of coagulation factors by adenoviruses may be of importance not only for the liver tropism seen when administering adenovirus vectors to the circulation but also during primary infections by wild-type viruses of their natural target cell types.

Quantification of Human β-Defensin-2 and -3 in Body Fluids: Application for Studies of Innate Immunity

Human beta-defensins (hBDs) are epithelial cell-derived antimicrobial and immunoregulatory cationic peptides. Our objective was to establish an analytical tool to quantify inducible hBD-2 and -3 in body fluids. We developed sandwich ELISAs using commercially available capture and detection antibodies and determined optimal assay conditions (with 250 mmol/L CaCl(2)) to overcome masking by endogenous components of body fluids. We used recombinant hBD as calibrators and for recovery testing. hBD-2 and -3 detection limits were approximately 75 ng/L and approximately 3 microg/L, respectively. Mean (SD range) values in saliva samples from healthy donors (n = 60) were 9.5 (1.2-21) microg/L for hBD-2 and 326 (50-931) microg/L for hBD-3. We did not detect hBD-3 in suction blister fluid (BF; n = 10) or bronchoalveolar lavage (BAL; n = 5) from healthy participants. We detected low hBD-2 peptide concentrations in BF and BAL, 0.16 (0.03-0.32) and 0.04 (0-0.049) microg/g total protein, respectively. We observed no correlation of hBD-2 in BF and saliva or BAL and saliva from the same person. In vaginal swabs from healthy women (n = 2), mean hBD-2 and -3 concentrations were 3.42 and 103 microg/g total protein, respectively. Cervicovaginal lavage from the same women contained mean concentrations of 1.46 and 55.5 microg/g total protein. These ELISA assays can measure inducible hBD peptide concentrations in body fluids by overcoming masking effects of anionic molecules. This approach may therefore be applicable for quantifying these peptides in health and disease.

Aluminum fluoride affects the structure and functions of cell membranes

No useful biological function for aluminum has been found. To the contrary, it might play an important role in several pathologies, which could be related to its interactions with cell membranes. On the other hand, fluoride is a normal component of body fluids, soft tissues, bones and teeth. Its sodium salt is frequently added to drinking water to prevent dental caries. However, large doses cause severe pathological alterations. In view of the toxicity of Al 3+ and F − ions, it was thought of interest to explore the damaging effects that AlF 3 might induce in cell membranes. With this aim, it was incubated with human erythrocytes, which were examined by phase contrast and scanning electron microscopy, and molecular models of biomembranes. The latter consisted of large unilamellar vesicles (LUV) of dimyristoylphosphatidylcholine (DMPC) and bilayers of DMPC and dimyristoylphosphatidylethanolamine (DMPE) which were studied by fluorescence spectroscopy and X-ray diffraction, respectively. In order to understand the effects of AlF 3 on ion transport (principally sodium and chloride) we used the isolated toad skin to which electrophysiological measurements were applied. It was found that AlF 3 altered the shape of erythrocytes inducing the formation of echinocytes. This effect was explained by X-ray diffraction which revealed that AlF 3 perturbed the structure of DMPC, class of lipids located in the outer monolayer of the erythrocyte membrane. This result was confirmed by fluorescence spectroscopy on DMPC LUV. The biphasic (stimulatory followed by inhibitory) effects on the isolated skin suggested changes in apical Cl − secretion and moderate ATPase inactivation.

Competitive adsorption of blood proteins on gamma-irradiated-polycarbonate films

Medical devices made of polycarbonates are generally sterilized by ionizing radiation. The effect of irradiation on the response of polycarbonate to various components of body fluids was studied in this work. Polycarbonate films prepared by solvent casting technique from various solvents were gamma-irradiated in the range of 0-200 kGy. Characterizations of the films were achieved by contact angle and water-uptake studies as well as atomic force microscope (AFM) images. It was found that gamma-irradiated films were more hydrophilic than unmodified films. AFM images showed that surface roughness increased with gamma irradiation. Protein adsorption experiments conducted with human plasma demonstrated that protein adsorptions drastically increased by increasing the applied irradiation dose. The unirradiated and gamma-irradiated films were contacted with human blood in in vitro systems. Loss of the blood cells and clotting times were followed. Loss of blood cells in the plasma contacting with gamma-irradiated films was negligible. Any significant change was also followed for clotting times with gamma-radiation dose.

The Pavia approach to clinical protein analysis.

The protein chemistry laboratory of the Pavia University Hospital is specialized in the study of monoclonal components in body fluids. It is closely connected with the research laboratory devoted to the structural and functional study of pathogenic proteins and with the Clinical Chemistry Central Laboratory. The analyses are performed on specific medical request. The analytical approach is mainly based on analysis of patient's serum and urine by high-resolution agarose gel electrophoresis and immunofixation with the possible addition of the quantification of specific proteins. The interpretation of the protein pattern and the final reporting result from the integration of the laboratory data with the clinical information. This labor-intensive approach requires skills in the performance of protein analysis and in the interpretation/referral phase, as well as close communication with the attending physician.

Glycoaminoglycan (GAG) deficiency in protective barrier as an underlying, primary cause of ulcerative colitis, Crohn's disease interstitial cystitis and possibly Reiter's syndrome

Summary Ulcerative colitis, Crohn's disease and interstitial cystitis share many common features, the most important of which is a defect in the glycoaminoglycan (GAG) defensive barrier. This defect allows penetration of toxins causing localized inflammatory response, followed by fibrosis and distant pathological changes, together with a myriad of biochemical and immunological changes. The latter has caused confusion as to etiology of the aforementioned disorders. This hypothesis is somewhat supported by the fact that agents such as glucosamine and pentosan polysulphate (Elmiron®) that replace the GAG layer, improve the conditions. The potential for extrapolation of this hypothesis to atherosclerosis and arthropathies exists.There is a great danger in modern medical research that if one misses the wood for the trees, one becomes hopelessly lost in the minutiae of research. At present, it is embarrassing that ulcerative colitis (UC), Crohn's (CR) and interstitial cystitis (IC) are the cause of a great deal of morbidity and occasionally mortality, yet after intensive research, the etiology and effective treatment eludes us. The research in the past has focused extensively on inflammatory response in the mucosal lining, and biochemical, infective and immunological changes in the serum. This has led to a vast array of research pathways that seem at the present time to be totally lost and, might I say, aimless in direction, as a cause for these conditions, that remain amongst the most imperically treated in modern medicine.Another possible syndrome in this class would be Reiter's, which has many features in common with the above.The basic tenet of a GAG deficiency hypothesis is that, as shown in Figure 1A, an intact GAG layer provides, firstly, a mechanical and electrostatic defence against penetration of infective agents, toxins, antigenic protein moieties, etc. and, secondly, the prevention of extravasation of body fluid components. A degraded GAG layer is the start of the disease cascade of the above group of illnesses.