C3 Consumption Research Articles

Reply to Fontes-Villalba et al.: On a reluctance to conjecture about animal food consumption

Fontes-Villalba et al. (1) correctly observe that carbon isotope ratios in tooth enamel do not speak directly to plant versus animal food ingestion. Carbon isotope ratio data are useful for quantifying the consumption of C3- or C4-derived carbon, whether it comes directly from C3 or C4 plants or indirectly through consumption of animals that eat those plants. Not only do we acknowledge as much in our recent series of papers, but we have made this point in print many times over the past two decades (ref. 2 and references therein).

Cardiac Surgery and C1-Inhibitor Deficiency

HEREDITARY ANGIOEDEMA (HAE) is a rare autosomal dominant disease (prevalence ≈ 1/100,000) caused by a deficiency in C1-esterase inhibitor (C1-inh) level (Type I HAE) or functional activity (Type II HAE).1–3 C1-inh regulates the activation of the classical complement pathway. In its absence, fraction C4 consumption occurs by autoactivation. C1-inh deficiency induces unregulated factor XII activation, which in turn activates the kallikrein-kinin system and plasmin. This results in excess bradykinin and increased capillary permeability with resultant local edema.

Seasonal dynamics of CO2 balance and water consumption of C3 and C4-type cover crops compared to bare soil in a suitability study for their use in vineyards in Germany and Argentina

Cover crops are used in vineyards to maintain soil structure, minimise soil erosion and optimise vineyard mechanisation, yet few data are available on water consumption and carbon dioxide assimilation (CO2). Whereas cover crop use is common in the grape growing regions of Germany it is uncommon in Argentina. To obtain some information on the suitability of a selected number of species in terms of carbon gain and water expenditure under the climatic conditions of Germany and Argentina, we first quantified evapotranspiration (EvT), CO2 assimilation and water use efficiency of Trifolium repens L. (white clover) (TR), Festuca arundinacea Schreb (tall fescue) (FE), Sorghum sudanense (Piper) Stapf (Sudan grass) (SO), and Digitaria californica (Benth) Henr. (Arizona cotton top) (DI) in a field trial in Geisenheim, Germany. We then compared the performance of TR, FE and DI together with Sorghum halepense L. (Johnson grass) (SH) in a pot experiment under the warmer climatic conditions of Mendoza, Argentina, where water supply could be controlled. In all cases bare soil served as a reference for soil water loss. Gas-exchange of plants and soil were measured with customised open system canopy chambers and an infrared gas analyser and water dynamics in the soil with soil moisture probes (field) or gravimetrically (pots). SO and TR were the two species that reached the highest degree of soil coverage in Germany and substantially reduced soil moisture in the first 0.40m of depth. TR showed high EvT and CO2 assimilation rates in both climates when water supply was sufficient and maintained relative high rates when water status was reduced under cool conditions in Germany, yet performed poorly when high temperatures occurred concomitant to low water supply in Argentina. DI, being a C4 species from arid regions, had difficulties to get established in the cool conditions of Germany and there was no clear tendency for more efficient water use than the other cover crops. However under the warm condition of Argentina, especially when water supply was reduced, DI displayed higher WUE which was entirely related to the maintenance of higher assimilation rates at substantially reduced EvT. FE being common in German vineyards had low assimilation rates combined with high evapotranspiration rates under Argentinian conditions and was very sensitive to water deficit in combination with high temperature.

Palaeobiology of the Medieval Population of Albano (Rome, Italy): A Combined Morphological and Biomolecular Approach

AbstractRecent years have seen increased interest in skeletal populations from the Imperial Roman Age in Italy, but much less is known about diet and standards of living in the subsequent medieval period. To fill this gap, we conducted a morphological analysis of human remains from Albano, an Italian town near Rome, as well as a stable isotope analysis of bone collagen to reconstruct diet. The sample was recovered from a Medieval cemetery (1040–1220 cal. yr. BP) located in the gardens of the historicalPalazzo Doria Pamphiliin Albano. A minimum number of 40 individuals (31 adults and 9 sub‐adults) were examined using standard methods. Though the general health status of the population was good, signs ofcribra orbitaliaand diffuse enthesopathies were noted during the morphological examination.Stable carbon and nitrogen isotope analyses of the bone collagen from 24 adult humans and three faunal bones indicate that the diet of the population may be described as predominantly terrestrial and C3‐plant based although the data for some of the individuals suggest a modest consumption of C4‐(millet) based or aquatic proteins. No evidence of significant dietary differences between the sexes was found.The comparison of the isotope data from Albano with those from populations recovered in the same region is consistent with a shift from a terrestrial, possibly plant foods‐dominated subsistence in the Early Middle Ages to a diet with a higher contribution from animal proteins, both terrestrial and aquatic, in the Later Middle Ages. Copyright © 2013 John Wiley & Sons, Ltd.

Oversulfated Chondroitin Sulfate Inhibits the Complement Classical Pathway by Potentiating C1 Inhibitor

Oversulfated chondroitin sulfate (OSCS) has become the subject of multidisciplinary investigation as a non-traditional contaminant in the heparin therapeutic preparations that were linked to severe adverse events. In this study, it was found that OSCS inhibited complement fixation on bacteria and bacterial lysis mediated by the complement classical pathway. The inhibition of complement by OSCS is not due to interference with antibody/antigen interaction or due to consumption of C3 associated with FXII-dependent contact system activation. However, OSCS complement inhibition is dependent on C1 inhibitor (C1inh) since the depletion of C1inh from either normal or FXII-deficient complement plasma prevents OSCS inhibition of complement activity. Surface plasmon resonance measurements revealed that immobilized C1inhibitor bound greater than 5-fold more C1s in the presence of OSCS than in presence of heparin. Although heparin can also inhibit complement, OSCS and OSCS contaminated heparin are more potent inhibitors of complement. Furthermore, polysulfated glycosaminoglycan (PSGAG), an anti-inflammatory veterinary medicine with a similar structure to OSCS, also inhibited complement in the plasma of dogs and farm animals. This study provides a new insight that in addition to the FXII-dependent activation of contact system, oversulfated and polysulfated chondroitin-sulfate can inhibit complement activity by potentiating the classical complement pathway regulator C1inh. This effect on C1inh may play a role in inhibiting inflammation as well as impacting bacterial clearance.

Common polymorphisms in the complement system and susceptiblity to bacterial meningitis

Risk factors for susceptibility to bacterial meningitis have been identified, but basic causes of inter-individual differences in susceptibility are largely unknown. To determine the effect of genetic variation in the complement system on susceptibility to bacterial meningitis we performed a prospective nationwide genetic association study in patients with community-acquired bacterial meningitis. We genotyped 17 common SNPs (minor allele frequencies >5%) in genes coding for complement components and evaluated functional consequences by measuring complement levels in the cerebrospinal fluid. From March 2006 to June 2009 we included 636 adults with community-acquired bacterial meningitis. DNA was available for 439 patients and 302 controls. Rs1047286 (Pro314Leu) in complement component 3 was associated with reduced susceptibility to bacterial meningitis after correction for multiple testing: the protective Leu/Leu genotype was found in 5 of 435 patients (1%) compared to 15 of 302 controls (5%; odds ratio [OR] 4.50, 95% confidence interval [CI] 1.62-12.50, p=0.0017). Rs1047286 is in strong linkage disequilibrium with Rs2230199 (C3 Arg102Gly), of which the Arg/Arg genotype was associated with higher CSF levels of C3 and lower levels of C5a and terminal complement complex (TCC; soluble C5b-9), indicating decreased consumption of C3 and less activation of the complement system. Rs1047286 was associated with susceptibility albeit not significantly after Bonferroni correction (OR 1.37, 95% CI 1.01-1.87; p=0.04). This study shows an association between a common single nucleotide polymorphism in C3 and susceptibility for community-acquired bacterial meningitis.

Overall Neutralization of Complement Factor H by Autoantibodies in the Acute Phase of the Autoimmune Form of Atypical Hemolytic Uremic Syndrome

Complement is a major innate immune surveillance system. One of its most important regulators is the plasma protein factor H (FH). FH inactivation by mutations or by autoantibodies is associated with a thrombotic microangiopathy disease, atypical hemolytic uremic syndrome. In this study, we report the characterization of blood samples from 19 anti-FH Ab-positive atypical hemolytic uremic syndrome patients collected at the acute phase of the disease. Analyses of the functional consequences and epitope mapping, using both fluid phase and solid phase approaches, were performed. The anti-FH Abs perturbed FH-mediated cell protection (100%), inhibited FH interaction with C3 (46%), and caused C3 consumption (47%). The Abs were directed against multiple FH epitopes located at the N and C termini. In all tested patients, high titers of FH-containing circulating immune complexes were detected. The circulating immune complex titers correlated with the disease stage better than did the Ab titers. Our results show that anti-FH autoantibodies induce neutralization of FH at acute phase of the disease, leading to an overall impairment of several functions of FH, extending the role of autoantibodies beyond the impairment of the direct cell surface protection.

Autoantibody stabilization of the classical pathway C3 convertase leading to C3 deficiency and Neisserial sepsis: C4 nephritic factor revisited

C3 deficiency is a rare disorder that leads to recurrent pyogenic infections. Here we describe a previously healthy 18 y/o Caucasian male with severe meningococcal disease. Total hemolytic activity was zero secondary to an undetectable C3. The C3 gene was normal by sequencing. Mixing the patient's serum with normal human serum led to C3 consumption. An IgG autoantibody in the patient's serum was identified that stabilized the classical pathway C3 and C5 convertases, thus preventing decay of these enzyme complexes. This autoantibody is an example of a C4 nephritic factor, with an additional feature of stabilizing the C5 convertase. Previous patients with C4 nephritic factor had membranoproliferative glomerulonephritis. Two years after presentation, this patient's C3 remains undetectable with no evidence of renal disease. We revisit the role of autoantibodies to classical pathway convertases in disease, review the literature on C4-NeF and comment on its detection in the clinical laboratory.

Changing cultures, changing cuisines: Cultural transitions and dietary change in iron age, roman, and early medieval croatia

Food is well-known to encode social and cultural values, for example different social groups use different consumption patterns to act as social boundaries. When societies and cultures change, whether through drift, through population replacement or other factors, diet may also alter despite unchanging resource availability within a region. This study investigates the extent to which dietary change coincides with cultural change, to understand the effects of large-scale migrations on the populations' diets. Through stable carbon and nitrogen isotope analysis of Iron Age, Roman, and Early Medieval human bone collagen, we show that in Croatia large-scale cultural change led to significant changes in diet. The isotopic evidence indicates that Iron Age diet consisted of C(3) foodstuffs with no isotopic evidence for the consumption of C(4) or marine resources. With the Roman conquest, marine resources were added to the diet, although C(3) foodstuffs continued to play an important role. In the Early Medieval period, this marine component was lost and varying amounts of C(4) foodstuffs, probably millet, were added to the otherwise C(3) diet. In both of these transitions it is likely that the changes in diet are related to the arrival of a new people into the area.

Behcet’s disease and focal segmental glomerulosclerosis: an unusual association

Behcet’s disease (BD) is characterised by oral aphthosis, uveitis, genital ulcers and skin lesions such as folliculitis and erythema nodosum [1, 2]. Glomerulonephritis is not a recognised feature of BD [3]. Only few cases of nonnephrotic focal segmental glomerulosclerosis (FSGS) associated with BD were reported [3, 4]. We report the Wrst case of nephrotic FSGS, associated with BD, successfully treated with cyclosporin (CsA). A 19-year-old man, admitted in our Department for nephrotic syndrome, showed oral ulcers, periodically recurring from at least 3 years, monocular right-sided uveitis, arthritis, leg thrombophlebitis and cutaneous vasculitis. There was no history of genital ulcers or erythema nodosum. HLA typing showed positive HLA B51, and the diagnosis of BD was made. He reported a history of mild hypertension from 2 years, well controlled through low-sodium diet and ramipril 5 mg/ day. Blood urea was 104 mg/dl and creatinine 1.9 mg/dl. Total serum proteins were 5.5 g/dl and serum albumin 2.9 g/dl. Twenty-four-hour urinary protein excretion was 4.2 g/day. Serum immunoglobulins were within normal levels. Serological tests for rheumatoid factor, antinuclear, anti-DNA and antineutrophil cytoplasmic antibody and cryoglobulins were negative. Urinalysis showed 4–6 leucocytes and 10–15 red blood cells/hpf. There was no evidence of complement factor C3 and C4 consumption. Chest X-ray, renal ultrasonography and ECG were normal. A percutaneous eco-guided renal biopsy was then performed. At light microscopy, 19 glomeruli were disclosed. Three glomeruli showed segmental sclerosis and collapse of the glomerular tuft with podocyte hyperplasia, suggesting the diagnosis of collapsing FSGS. There was no mesangial proliferation. Both IgM (1+) and C3 (1+) in mesangial area and IgG (1+) in glomerular basement membranes were detected by immunoXuorescence. Oral prednisone (1 mg/kg/day, alternate day), CsA 200 mg/day, ramipril 10 mg/day, furosemide 100 mg/day and atorvastatin 40 mg/day were started. At 1 month, his 24-h proteinuria was 425 mg and his serum creatinine was 1.3 mg/dl. At 6 months, his 24-h proteinuria was 489 mg and his serum creatinine was 1.3 mg/dl. Oral steroids were withdrawn from therapy at 6 months. This total remission remained stable at 4 years from the biopsy, with a 24-h proteinuria of 356 mg and a serum creatinine of 1.2 mg/dl. Daily treatment with CsA 100 mg, ramipril 10 mg and atorvastatin 40 mg was maintained. This is the only case reported with FSGS at nephrotic presentation in BD in our knowledge. He responded with total remission to a treatment with CsA, steroids, ACE inhibitors and statins. Dermatologists, rheumatologists and ophthalmologists taking care of BD patients should be aware about potential renal involvement and therefore periodic urine examination and serum creatinine monitoring should be part of the management protocol. A. Granata (&) Department of Nephrology and Dialysis, AOU “Policlinico Vittorio Emanuele”, Catania, Italy e-mail: antonio.granata4@tin.it

Formulations combining CpG containing oliogonucleotides and poly I:C enhance the magnitude of immune responses and protection against pancreas disease in Atlantic salmon

Both CpG oligodeoxynucleotides and double-stranded RNA (poly I:C) have documented effects as treatments against several viral diseases in fish. However, as stand-alone treatments their effects have been modest. We have tested here whether CpG and poly I:C, alone or in combination induce protection against Salmonid Alphavirus (SAV), the causative agent of pancreas disease in Atlantic salmon. Our results revealed a significant reduction of viraemia 2 weeks after ip injection of the combined treatment and 1 week after challenge with SAV subtype 3, followed by reduced SAV induced heart pathology 3 weeks later. The SAV titers in blood samples from the combination group were lower as compared to single treatments with either CpG or poly I:C. Surprisingly, reduced SAV levels could also be found in fish as long as 7 weeks after receiving the combination treatment. The expression of IFNγ and to a lesser extent IFNa and Mx was up-regulated in head kidney and spleen 5 days after the fish had been treated with CpG and poly I:C. Furthermore, the complement factor C4 was depleted in serum 8 weeks post treatment, suggesting complement activation leading to C4 consumption. We hypothesize that the CpG/poly I:C-induced protection against SAV3 is mediated by mechanisms involving type I and type II IFN induced antiviral activity and complement mediated protective responses.

Evidence of impaired sense of smell in hereditary angioedema

Hereditary angioedema (HAE) is an autosomal-dominant disorder resulting from C1-inhibitor (C1INH) deficiency. Smell impairments were found in patients affected with systemic lupus erythematosus, that, similarly to HAE, is characterized by the activation of the classical complement pathway with C4 consumption. In this study, we aimed at evaluating the sense of smell in patients with HAE. Thirty patients with HAE and 30 healthy age- and sex-matched controls were evaluated for olfactory functions using the 3-stages Sniffin'-Sticks kit (threshold, discrimination, and identification [TDI]). TDI scores were analyzed according to complement levels (C1INH, C3, C4 and CH50), Beck depression inventory (BDI-II) and danazol treatment. A significant decrease in olfactory function was observed in patients affected with HAE compared with controls in total TDI score (P < 0.001), and in the discrimination (P < 0.001) and identification scores (P = 0.012). Anosmia was present only in patients with HAE (3.3%) who also exhibited more frequently hyposmia (53.3%vs 3.3%, P < 0.0001). Complement levels were reduced in patients with HAE. C4 serum levels showed positive correlation with total TDI score (P < 0.001), and with discrimination (P = 0.002) and identification (P = 0.011) scores. CH50 complement levels showed positive correlation with total TDI score (P < 0.001), and with threshold (P = 0.002) and discrimination (P = 0.011) scores. Sex, age, danazol treatment, BDI-II scores were not different between the patients and controls and did not influence TDI scores significantly. Evidence for an impaired sense of smell was found in patients with HAE. The reduction in olfactory function in these cases seems to correlate with complement C4 and CH50 levels. Immune and genetic mechanisms might play a role in this defect.

A structural basis for Staphylococcal complement subversion: X-ray structure of the complement-binding domain of Staphylococcus aureus protein Sbi in complex with ligand C3d

The structure of the complement-binding domain of Staphylococcus aureus protein Sbi (Sbi-IV) in complex with ligand C3d is presented. The 1.7Å resolution structure reveals the molecular details of the recognition of thioester-containing fragment C3d of the central complement component C3, involving interactions between residues of Sbi-IV helix α2 and the acidic concave surface of C3d. The complex provides a structural basis for the binding preference of Sbi for native C3 over C3b and explains how Sbi-IV inhibits the interaction between C3d and complement receptor 2. A second C3d binding site on Sbi-IV is identified in the crystal structure that is not observed in related S. aureus C3 inhibitors Efb-C and Ehp. This binding mode perhaps hints as to how Sbi-IV, as part of Sbi, forms a C3b–Sbi adduct and causes futile consumption of C3, an extraordinary aspect of Sbi function that is not shared by any other known Staphylococcal complement inhibitor.

Properdin homeostasis requires turnover of the alternative complement pathway

Properdin is a plasma protein and is also released from neutrophil granules following stimulation. At inflammatory sites it can bind bacteria and apoptotic bodies to trigger alternative pathway (AP) activation. Principles governing properdin homeostasis are unknown. We monitored properdin during AP activation and in complement-deficient mice. There was a >90% reduction of properdin in the Crry single-knockout mice (Crry SKO). These membrane complement regulatory protein-deficient mice feature accelerated AP turnover, leading to reduced C3 and fB. Injecting cobra venom factor into wild-type mice activated the AP and led to the consumption of C3, fB, and properdin. However, and unexpectedly, properdin was also deficient in C3(-/-), fB(-/-), and fD(-/-) mice. It was present in C1q(-/-), C4(-/-), and C5(-/-) mice. These findings implicate AP turnover in the maintenance of basal levels of properdin in the blood. To explore the mechanism, classical pathway-activating immune complexes were infused. Within 10 min, properdin was partially restored in fB(-/-) but not in C3(-/-) mice. Markedly reduced properdin in mice deficient in an AP component and its partial restoration by activating C3 suggest a requirement for continuous C3 activation via AP tickover to maintain properdin homeostasis. The mechanism underlying this C3-dependent process was not identified. Engagement of C3a and C5a receptors was ruled out. These findings represent an instructive example of how a positive regulator of an innate immune recognition and effector pathway is controlled. A rationale for such a means to supply properdin for immune reactions is proposed.

Complement factor 3 deficiency attenuates hemorrhagic shock-related hepatic injury and systemic inflammatory response syndrome

Although complement activation is known to occur in the setting of severe hemorrhagic shock and tissue trauma (HS/T), the extent to which complement drives the initial inflammatory response and end-organ damage is uncertain. In this study, complement factor 3-deficient (C3(-/-)) mice and wild-type control mice were subjected to 1.5-h hemorrhagic shock, bilateral femur fracture, and soft tissue injury, followed by 4.5-h resuscitation (HS/T). C57BL/6 mice were also given 15 U of cobra venom factor (CVF) or phosphate-buffered saline injected intraperitoneally, followed by HS/T 24 h later. The results showed that HS/T resulted in C3 consumption in wild-type mice and C3 deposition in injured livers. C3(-/-) mice had significantly lower serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) and circulating DNA levels, together with much lower circulating interleukin (IL)-6, IL-10, and high-mobility group box 1 (HMGB1) levels. Temporary C3 depletion by CVF preconditioning also led to reduced transaminases and a blunted cytokine release. C3(-/-) mice displayed well-preserved hepatic structure. C3(-/-) mice subjected to HS/T had higher levels of heme oxygenase-1, which has been associated with tissue protection in HS models. Our data indicate that complement activation contributes to inflammatory pathways and liver damage in HS/T. This suggests that targeting complement activation in the setting of severe injury could be useful.

A Favorable 3-Year Outcome of Kidney Transplantation in Atypical Hemolytic Uremic Syndrome Associated With a Factor H Mutation: Case Report

Complement factor H (CFH)-associated hemolytic uremic syndrome (HUS) is a genetic form of atypical HUS characterized by deficient CFH levels or activity, which cause a disorder of the regulation of the alternative pathway, leading to uncontrolled complement activation. This genetic disorder, which frequently leads to end-stage renal failure, often recurs in kidney transplants, resulting in the poorest graft outcomes among all atypical HUS forms, due to a mutation in genes encoding complement components and regulatory proteins. Herein we have report our experience with a 40-year-old woman, suffering from a clearly defined sporadic form of genetic atypical HUS, consisting of a heterozygous missense mutation in factor H gene. She underwent cadaveric kidney transplantation. At the moment of surgery she displayed positive hemolysis indices and C3 consumption. A calcineurin inhibitor (CNI)-free immunosuppressive regimen was based on sirolimus, mycophenolic acid and steroids after basiliximab induction. An early and intense prophylactic course of plasma exchange (PE), and fresh frozen plasma (40 mL/kg) was prescribed, starting before surgery and continuing daily for the first week. The frequency of PE slowly reduced over the following 2 weeks. After that, just plasma infusion at the same dose was performed once a week until 12 weeks after transplantation. There was prompt graft function and in third week there were no signs of hemolysis or of C3 consumption. More than 3 years after transplantation, the graft is still functioning well and there was no recurrence. In our opinion, this case indicates that, although evidence is lacking, avoidance of CNI and intensive prophylactic plasma therapy are essential to achieve good results in this peculiar type of kidney transplantation. Nevertheless, controlled, prospective studies are necessary to establish the actual role of these two therapeutic procedures in renal transplantation of patients with CFH-associated HUS.

Anti-factor B autoantibody in dense deposit disease

Dense deposit disease (DDD), also known as membranoproliferative glomerulonephritis type II, is a rare kidney disorder that is associated with dysregulation of the alternative pathway of complement. Autoantibodies against the C3bBb convertase termed C3 nephritic factor are common in DDD patients. Here we report an autoantibody that binds to complement factor B in a DDD patient who was negative for C3 nephritic factor. This anti-factor B autoantibody recognized an epitope within the Bb fragment and was able to bind to the C3bBb convertase. Upon binding, the anti-factor B autoantibody stabilized the convertase against both intrinsic and factor H-mediated extrinsic decay and thus enhanced C3 consumption. Functional analyses demonstrated that, in contrast to C3 nephritic factor, the anti-factor B autoantibody inhibited complement-mediated lysis in vitro due to inhibition of the C5 convertase and the terminal complement pathway. Analysis of C5a plasma levels indicated that not all C5 convertases are inhibited by the autoantibodies in the patient in vivo. Antigen array experiments confirmed the presence of anti-factor B autoantibodies and also revealed complement activating anti-C1q antibodies in the patient's plasma. In summary, the present report describes a new autoantibody in DDD that binds to factor B and to the alternative pathway C3 convertase and alters the kinetics of complement activation and regulation.

Stable carbon isotope reconstructions of diet and paleoenvironment from the late Middle Pleistocene Snake Cave in Northeastern Thailand

Thailand's geographical location in the tropics and almost complete, relatively uninterrupted forest cover makes it valuable for paleodiet and paleoclimate research. We present the first dietary and environmental reconstructions in Northeastern Thailand, using stable isotope abundances in mammalian tooth enamel from the late Middle Pleistocene locality, Tham Wiman Nakin (Snake Cave), which reflect a much higher (over 70%) than modern (13%) occurrence of C4 plants. Bovids and cervids appear to have had almost entirely a C4 plant diet. Carnivores consumed a mixture of C3 (suids) and C4 (bovids, cervids) consumers. Rhinoceroses and orangutan appear to have maintained their preference through time for forested or open C3 environment, respectively. (13)C/(12)C from bone bioapatite, horn and hair of modern Southeast Asian mammals almost exclusively demonstrate C3 vegetation dominance. C4 consumption is rare in analysed modern species and it could be related to anthropogenic influences such as ingestion of domestic crops or livestock. Interesting implications emerge in the C4 vegetation distribution in southern Eurasian ecosystems, indicating that Southeast Asia, south of the Tibet, could be part of the global C4 vegetation spread, which occurred around 7 Ma. However, the C4 percentage in ecosystems varied geographically. Despite modern reversal towards C3 habitats due to factors such as increasing CO(2), we think that anthropological influences may be responsible for habitat and dietary changes in extant species. Bovids demonstrate the most significant shift in diet and habitat through time, from C4-dominated open habitats to C3-dominated habitats indicative of dense forest understory.

Stable isotope analysis of modern human hair collected from Asia (China, India, Mongolia, and Pakistan)

We report isotopic data (delta(2)H, delta(18)O n = 196; delta(13)C, delta(15)N n = 142; delta(34)S n = 85) from human hair and drinking water (delta(2)H, delta(18)O n = 67) collected across China, India, Mongolia, and Pakistan. Hair isotope ratios reflected the large environmental isotopic gradients and dietary differences. Geographic information was recorded in H and O and to a lesser extent, S isotopes. H and O data were entered into a recently developed model describing the relationship between the H and O isotope composition of human hair and drinking water in modern USA and pre-globalized populations. This has anthropological and forensic applications including reconstructing environment and diet in modern and ancient human hair. However, it has not been applied to a modern population outside of the USA, where we expect different diet. Relationships between H and O isotope ratios in drinking water and hair of modern human populations in Asia were different to both modern USA and pre-globalized populations. However, the Asian dataset was closer to the modern USA than to pre-globalized populations. Model parameters suggested slightly higher consumption of locally produced foods in our sampled population than modern USA residents, but lower than pre-globalized populations. The degree of in vivo amino acid synthesis was comparable to both the modern USA and pre-globalized populations. C isotope ratios reflected the predominantly C(3)-based regional agriculture and C(4) consumption in northern China. C, N, and S isotope ratios supported marine food consumption in some coastal locales. N isotope ratios suggested a relatively low consumption of animal-derived products compared to western populations.

A Large Family with a Gain-of-Function Mutation of Complement C3 Predisposing to Atypical Hemolytic Uremic Syndrome, Microhematuria, Hypertension and Chronic Renal Failure

Atypical hemolytic uremic syndrome (aHUS) is associated with mutations in genes encoding complement-regulatory proteins factor H, I and B and membrane cofactor protein. Recently, heterozygous gain-of-function mutations in the complement C3 gene have been found in patients with aHUS. A large family with a C3 R570Q mutation is described. Clinical and laboratory findings of carriers of the mutation and unaffected family members are reported. The index patient suffered from recurrent aHUS at age 22 and developed end-stage renal failure. Of 24 family members, nine harbored the C3 R570Q mutation. Carriers showed reduced or borderline C3 levels. Arterial hypertension was found in six family members, microhematuria in five and chronic kidney disease stage 3 in two elderly carrier patients. Despite marked consumption of C3, serum terminal complement complex levels were not elevated in carriers compared with other family members. The penetrance of the C3 R570Q mutation to induce aHUS is incomplete and lower compared with mutations in other genes predisposing to the disease. The mutation is possibly also associated with hypertension, hematuria and chronic kidney disease, all of which may represent consequences of long-term complement activation in the renal vasculature.