Thus, α-gliadin genes can be assigned to specific chromosome loci

Thus, α-gliadin genes can be assigned to specific chromosome loci according to their marked genomic differences [12] and [13]. Further analysis of group 6 nulli-tetrasomic lines of Chinese Spring confirmed the reliability of such assignment methods for α-gliadin genes [23]. In conclusion, α-gliadins not only play a major role in determining gluten quality, but comprise the major source of toxicity for CD patients, given that they contain most of the main toxic components. In addition, this multigenic family encodes extensive buy ABT-737 allelic variation that has been shown to be closely associated with flour quality [24] and [25]. Screening of new

allelic variants with specific profiles of α-gliadins from common wheat cultivars with good quality or from other valuable Triticeae species may accordingly aid in exploring

gene resources both for quality improvement and potential CD prevention. The objective of the current study selleck was to clone and characterize the novel full-ORF α-gliadin genes from common wheat cultivar Zhengmai 004, one of the major cultivars sown on a large scale in the weak-gluten wheat growing areas of China owing to its good quality and high and stable yield. To shed light on the structure–function relationships of a single α-gliadin gene, the prokaryotic expression in Escherichia coli of two genes differing in the number of cysteine residues was investigated by SDS-PAGE and Western blotting. Finally, the secondary structures of the full-ORF genes cloned in this study and other genes in the public database GenBank derived from common wheat and its relatives were

predicted and the typical secondary structure of α-gliadins was summarized. Seeds of Zhengmai 004 were kindly provided by Professor Hu Lin from the Wheat Research Institute of Henan Academy of Agricultural Sciences, Zhengzhou, China. Genomic DNA was extracted from young leaves of 10–20 wheat seedlings grown in the greenhouse, using the cetyltrimethyl ammonium bromide (CTAB) procedure. A pair of degenerate primers (F: 5′-GGA TCC ATG AAG ACC TTT CTC ATC CT-3′; R: 5′- AAG CTT TCA GTT RGT ACC GAA GAT GCC-3′) with respectively Bam H I and Hind III sites (underlined) at the 5′-end of each primer was designed according to the majority of the published open reading frame (ORF) sequences of α-gliadin genes in Clomifene GenBank. PCR was performed using LA Taq (TaKaRa, Dalian, China) with GC buffer (1 unit) in a 20-μL reaction volume containing approximately 50 ng of genomic DNA, 100 μmol L− 1 of each dNTP, and 0.5 μmol L− 1 of each primer. PCR cycling was at 94 °C for 4 min followed by 10 cycles of 94 °C for 30 s, 62 °C (Tm + 4 °C) for 45 s, 72 °C for 60 s, then 22 cycles of 94 °C for 30 s, 58 °C for 45 s, 72 °C for 60 s, and a final extension at 72 °C for 15 min. PCR products were separated on 1% agarose gels and the single target fragment was purified from the gels using Gel Extraction Kit Ver 2.0 (TaKaRa, Dalian, China).

Neurological assessment (including MRC scale) at the time of myos

Neurological assessment (including MRC scale) at the time of myosonology showed clinical features of myopathic syndrome more pronounced for distal leg muscles in all patients. Normal conduction velocities of the fibular, tibial, median nerves and myogenic changes of distal calves and hand muscles were found by electromyography. An advanced muscular dystrophy was proved by muscle biopsy, performed in the patients with VCPDM and TMD ZD1839 ( Table 1). Triceps surae muscles were evaluated in a lying position by using a special probe for 3D/4D real time

imaging (Logic 7, GE). The transverse diameter of both TS heads in longitudinal Sirolimus research buy plan, the angle of inclination of the muscle fibers towards the surface of the aponeurosis and 3D/4D imaging of calf architectonics were evaluated in rest and during maximal plantar flexion (PF). The results were compared to myosonograms of 3 age- and sex-matched healthy controls. The normal TS myosonogram is demonstrated in Fig. 1.

The whole muscle is enveloped by hyperechoic epimysium. The muscle fibers are hypoechoic and grouped in fascicles, divided by hyperechoic septs of fibrous and fat tissue of the perimysium. In a longitudinal B-mode image the perimysium is depicted as oblique parallel hyperechoic lines. The PF causes calf muscle contraction that increases the transverse muscle diameter and the angle of muscle fiber towards the aponeurosis. The 4D ultrasound imaging shows a reticular TS architectonics despite the muscle activity, age and sex of healthy controls. Its hypoechoic areas increase during PF, Orotic acid due to thickening

of the contracted muscle fibers. Compared to healthy controls all patients with DM had a reduced transverse TS diameter and decreased muscle contractility. The muscle fibers were inclined and their orientation was under a smaller angle towards the aponeurosis during rest and PF (Fig. 2). The normal reticular muscle structure was replaced by granular myoarchitectonics – a combination of spot-like hypo- and hyperechoic areas on 4D ultrasound imaging was found in association with the degree of muscle atrophy, fat tissue infiltration and fibrosis. The hyperechoic areas had a tendency of fusing in the patient with HIBM2 (Fig. 3). Distal myopathies are a group of genetically and clinically heterogeneous disorders classified into one broad category, due to the presentation of weakness involving distal skeletal muscles of upper and lower limbs.

Eutrophication can also increase the severity of diseases (Bruno

Eutrophication can also increase the severity of diseases (Bruno et al., 2003) and lead to competitive advantage for macroalgae that respond by rapid growth, smothering corals or blocking light (Lapointe, 1997 and Walker and Ormond, 1982), although evidence for different trajectories also exists (McCook, 1999a and McCook, 1999b). Sediments that are influenced by outflow from industrial areas can

contain relatively high levels of lead, cadmium, copper, tin, nickel and iron (Amin et al., 2009 and Todd et al., 2010). In particular, copper is known to inhibit coral recruitment, fertilisation and development (Reichelt-Brushett and Harrison, 2005 and Negri and Hoogenboom, 2011). Light-enhanced calcification is responsible for most of the skeletal growth of reef-building corals (Goreau, 1959). Low light decreases calcification in zooxanthellate scleractinian corals, being approximately three times lower in darkness than in light (Kawaguti and Sakumoto, 1948 and Gattuso et al., 1999). Titlyanov (1991), however, noted that enhanced utilisation of light by zooxanthellae in three stony corals can result in stable levels of primary production in a wide light range (20–90% PAR). Low light GPCR & G Protein inhibitor levels may also inhibit the development

of coral larvae (Rogers, 1990). Similar patterns of photo-acclimation (through photophysiological adaptations) across gradients of increased turbidity have been demonstrated by Hennige et al., 2008 and Hennige et al., 2010. Although certainly also related to a variety of other environmental factors, species diversity of corals generally tends to decrease sharply with increasing (chronic) turbidity (Rogers, 1990, Becking et al., 2006 and Cleary et al.,

2008). Long-term turbidity stress can shift the species composition of reefs through the death of more light demanding corals and the subsequent replacement by usually deeper-living, more shade-tolerant ones at certain depths (Pastorok and Bilyard, 1985). Dikou and van Woesik (2006b) noted in Singapore the occurrence of deeper-water genera such as Merulina, Pachyseris and Mycedium found in relatively Immune system shallow (3–4 m) depths was most likely due to high turbidity levels. Also in Singapore, Goh et al. (1994) considered the sediment-impacted light environment to be the main factor controlling coral colony form. Foliose forms tended to dominate the shallow reef with more massive and encrusting forms found deeper. Corals can react either actively or passively to sediments, which in many ways defines their capability to withstand prolonged sedimentation. Passive shedding refers to corals taking advantage primarily of their shape to allow increased runoff of sediment, to maintain parts of the corallum above sediment, or to use water currents to remove accumulated sediment (Stafford-Smith and Ormond, 1992, Stafford-Smith, 1993, Riegl, 1995, Riegl et al., 1995 and Sanders and Baron-Szabo, 2005).

Our results support our hypothesis that

high Se intake or

Our results support our hypothesis that

high Se intake or status negatively impacts basal blood glucose management. Contrary to our hypothesis and previous reports demonstrating a positive effect of an HIF diet on glucose management, we found that HIF intake did not attenuate the increased fasting blood glucose that resulted from SMSC supplementation. Interestingly, although not statistically significant, there was a tendency for improved glucose tolerance in animals that were given both elevated SMSC and HIF compared with SMSC alone. Furthermore, both Se and IF have been reported to affect AMPK activation and thus cause changes in glucose management. Contrary to our original hypothesis, we did not observe a change in basal AMPK activation with SMSC supplementation, but we did observe a reduction with increased dietary IF. Selenium is an essential component of enzymes BIBF 1120 price critical to antioxidant defense. Although the precise mechanisms are not completely understood, high Se intake or status has been reported to reduce the risk of developing prostate and other cancers. However, in contrast to its chemopreventive

effects, high Se intake or status may also have a negative impact on blood glucose management. The effect that supplemental Se has on blood glucose is clearly dependent on the chemical form of Se administered [10], [13] and [22]. We supplemented mice with SMSC, an organic form of Se that Tacrolimus datasheet is abundant in foods high in Se and has a high bioavailability. Although our results contrast with those seen from increased intake of sodium selenate [10] and [11],

our findings are consistent with observational studies that have found a correlation between increased serum Se and increased incidence of type 2 diabetes [23] and [24]. Acetophenone A small increase in the risk of developing type 2 diabetes was also found after supplementation of selenomethionine in the large, randomized, controlled SELECT [14]. In addition to the results of the SELECT trial, the randomized, controlled trial reported by Stranges et al showed a significant increase in the incidence of type 2 diabetes resulting from supplementation of 200 μg Se daily in a high-Se brewer’s yeast tablet, which provides Se in multiple chemical forms [24]. However, in that study, the increased risk from Se supplementation was confined to those in the highest tertile of baseline plasma Se (>121.6 ng/mL). Those who began the study with lower plasma Se concentration experienced no increase in risk for T2D from consuming high-Se yeast. The mechanisms by which increased dietary IF improve glucose management are not clear. Cederroth et al [17] have reported that increased IF cause increased activation of AMPK in peripheral tissues. As noted above, one of the proposed mechanisms by which elevated Se negatively impacts insulin sensitivity is by reducing AMPK activation [15].

Additionally, and what I think most important, no hurricanes stru

Additionally, and what I think most important, no hurricanes struck the Keys in the 27-year-period between Betsy in 1965 and Andrew in 1992. Thankfully, Andrew missed the heart of the Keys. Burger Kings, McDonalds, gas stations, and marinas popped up during the later

part of the 1970s. However, the biggest social and monetary change occurred when an exotic grouper appeared: “square grouper,” the local name for bales of marijuana. Pot, smuggling, and later cocaine, brought sudden wealth, and almost overnight previously poor lobster fishermen were driving Mercedes. Some purchased selleck chemicals fleets of boats and thousands of traps. Motels and marinas grew larger and property values skyrocketed. Many boats moored in the newly built Port Largo canal system sported noticeably high PARP inhibitor review water lines. Boats with waterlines below the surface were a dead giveaway to contraband loaded below decks. Scruffy young sail boaters could be seen purchasing burgers at the nearby Burger King with hundred dollar bills, and small planes landed night and day on the landing strip that paralleled the main channel to the Port Largo. Today, expensive homes dot what was then the runway. Homes, property, and boats were being

bought with suitcases of hard cash, while beer trucks transported weed northward on US 1. Meanwhile illegal aliens literally floated in on rafts and makeshift boats, leading Immigration and Customs agents to set up roadblocks Sclareol on US 1. They were usually right next to the Last Chance Bar and Grill. That was before US 1 was relocated to its present location east of the Last Chance. Inspecting car trunks for illegal aliens revealed the true extent of drug smuggling, so periodic

roadblocks persisted. These roadblocks of course impacted tourism—and smuggling, leading to establishment of the so-called Conch Republic on April 23, 1982. Creating the Republic and threatening to secede from the Union was a publicity stunt, but the term Conch Republic stuck and proudly remains today. To avoid being caught at the roadblock, smugglers could telephone the Last Chance Bar (they posted their phone number on a chalk board) and learn if one was in place. Too many Keys politicians and public employees found easy money irresistible. Some roads to nowhere were constructed. The one on Sugarloaf Key now has a gate to prevent access. It was always covered with skid marks where small planes landed to unload. The Keys were a very different place worthy of many Jimmy Buffett songs. “A pirate turns 40” was popular. The exact dates escape me but a Supreme Court decision limited the State’s offshore jurisdiction to 3 miles on the Atlantic side of the Keys. Pennekamp State Park could no longer protect the best reef areas farther offshore. This change in State jurisdiction provided an opportunity for NOAA’s new Marine Sanctuary Program to collaborate with the State.

Blood samples were collected 1 h later and serum creatine kinase

Blood samples were collected 1 h later and serum creatine kinase (CK) activity was measured using Merck Granutest 2.5. Concentrations of 0, 25, 50, and 100 μg of purified 59/2-E4 mAb were incubated with 5 μg of B. atrox venom and injected i.d. into the shaved back of three Swiss mice. After 30 min, animals were euthanized and the size and intensity of subcutaneous hemorrhage

in injected areas was estimated. 3.5 mg samples of purified mAb 6AD2-G5 were preincubated with 150 μg of venom for 30 min at ambient temperature and i.p. injected into five Swiss mice (18–22 g). One hour after inoculation, the tips of tails were cut and immersed in 10 mL of distilled water until bleeding stopped (Assafim et al. 2006; Greene et al. 2010). learn more The optical density of samples was determined in a spectrophotometer at 410 nm. In addition, 500 μL of horse F(ab′)2 bothropic antivenom was used as positive control group, whereas venom plus saline was injected into the mice as negative control. Groups of five Swiss mice (18–20 g) were injected i.p. with selleck chemicals 500 μL saline containing 5 mg 59/2-E4, 5 mg A85/9-4, and 3.5 mg 6AD2-G5 mAb. After 30 min, mice were challenged s.c. with 350 μg of crude venom. Controls were injected i.p. with 500 μL saline and challenged s.c. with 350 μg

of venom. In another experiment 10.5 mg of mAbs (3.5 mg of each mAbs) were incubated with 200 μg of venom for 30 min at 37 °C followed i.p. injection into the mice. The control group received 200 μg of venom. Survival/death rates were recorded at 24 and 48 h. A mixture containing 3.45 mg each of mAb 59/2-E4, A85/9-4, and 6AD2-G5 incubated with 200 μg of venom was injected i.p. in groups of six Swiss mice. Controls received only saline and venom. After 2, 24, and 48 h, two mice from each group were euthanized by CO2 inhalation and their tissues and organs removed and fixed in 10% neutral p-formaldehyde. Tissues were dehydrated in ascending concentrations of ethanol (70–100%) and embedded in paraffin

using an automatic tissue processor (TP 1020, Leica, Germany). Docetaxel in vitro Then, 5 μm sections were stained with hematoxylin-eosin and tissue sections were observed using a digital image analysis system coupled to a microscope (Zeiss axioplan/axiocam, Germany). We evaluated the lethality neutralization by monoclonal antibodies against three major toxic components of B. atrox venom to test the prospects of developing bothropic antivenom based on monoclonal antibodies. General features of purified mAb specific to serineproteinase (thrombin-like 6AD2-G5 clone), PLA2 (A85/9-4 clone), and hemorrhagin (Zn-metalloproteinase 59/2-E4 clone) are shown in Fig. 1. When submitted to SDS-PAGE analysis, all three mAb preparations demonstrated two major protein bands, one of around 55 kDa and one of approximately 29 kDa, suggestive of immunoglobulin heavy and light chains, in addition to several minor contaminant bands ( Fig. 1A).

These results indicate that dd-PCR is more sensitive for the dete

These results indicate that dd-PCR is more sensitive for the detection and quantification of DNA from digester samples, which is consistent with a observation by Kim et al. [8]

that dd-PCR was more sensitive for quantifying DNA from soil than qPCR. PCR inhibitors co-extracted with nucleic acids from environmental samples can adversely affect qPCR quantification [16]. dd-PCR may be less sensitive to PCR inhibitors than the qPCR because the post-PCR quantification regime after 40 cycles can tolerate wide variations in PCR amplification efficiencies [6] and [12]. The technologies detected five groups: msar, msa, selleck compound mcp, msp, and mcr7 ( Figs. 1a–e), and therefore, they were compared based on these groups. T-test revealed that the technologies identically indicated the digesters in which the target groups were most abundant at p < 0.05. Both technologies also showed the same order among the digesters in order of abundance of msa, mcp, msp, and mcr7 (p < 0.05). In the case of msar, both technologies showed that it was much greater in digester C than in digesters A and B, and dd-PCR showed it was greater in digester A than in digester B, while qPCR showed the opposite (p < 0.05). The linear regression (y = ax + b) was conducted in order to determine whether or not there were quantitative agreements between the dd-PCR and qPCR measurements. Similar to previous observations

showing quantitative agreements between both technologies [4] and [15], there were R2 values ranging from 0.59–0.98 in all of the groups ( check details Fig. 1). However, slope values substantially varied between the groups. Both technologies quantitatively agreed, although their quantitative differences were quite varied. In order to determine whether or not both datasets represent similar relationships among the digester communities, principal component analysis (PCA), a multivariate approach to compare microbial communities, was performed using CANOCO version 4.5 [18]. The PCA plot of dd-PCR shows Thalidomide that the first and second principal component axes account for 88.3 and 11.1% of the compositional variance in the data,

respectively (Fig. 2a), whereas that of qPCR shows that the first and second axes account for 98.1 and 1.9% (Fig. 2b). Both plots indicate a substantial difference in the community composition among the digesters, and exhibit similar levels of differentiation among the communities. Both plots also indicate that operational temperature (from 38 to 52.5 °C) coincides with the score of the first axis (from approximately −0.5 to 1.0). Both plots indicate that the community of the thermophilic digester C was distinct from those of the mesophilic digesters A and B, primarily because msar dominated the C community. The msar (Methanosarcina) abundance increased along with the temperature, since Methanosarcina is better established in thermophilic regimes than in mesophilic regimes [1].

Catalog #P6181) was added at 1:20 ratio of enzyme to substrate an

Catalog #P6181) was added at 1:20 ratio of enzyme to substrate and incubated at 37 °C for ∼24 h Rat.

The patch clamp method (Hamill et al., 1981) was used to trace and record, ionic currents from heterologous expression systems over-expressing a recombinant channel. GW-572016 cost NaV1.3 channels were expressed in HEK-293 cells as described (Cummins et al., 2001). There is some degree of endogenous expression of NaV channels in HEK cells, but their contribution in comparison to exogenously expressed channels is usually minute (Moran et al., 2000). Rat NaV1.8 channels were expressed in ND7-23 cells by using conventional transient or stable transfections as described (Zhou et al., 2003, John et al., 2004, Jarvis et al., 2007 and Zimmermann et al., 2007). HEK cells stably expressing human NaV1.3, human NaV1.8 and human NaV1.7 channels were purchased from Scottish Biomedical (Glasgow, UK). Human NaV1.5 channels were expressed in HEK-293 cells as described

(Van Bemmelen et al., 2004). The patch clamp set up included amplifier and digitizer (Axopatch 200B and DIGDATA 1322A, FGFR inhibitor Axon instruments, USA), microscope (Nikon ECLIPSE 100) and micromanipulator (MP-225-Sutter Instrument Co., USA). Recording pipettes were pulled from Borosilicate glass tubes (Sutter Instrument co., USA). Cells were always perused with control extracellular solutions and changing to reagents containing solutions was performed using ValveLink 16 (Automate Scientific Inc. Berkeley, USA) and a peristaltic pump (Ismatec, Wertheim, Germany) perfusion system. Intracellular (pipette) solution (in mM): 120 CsF, 10 NaCl, 10 TEA-OH, 1 MgCl2, 1 CaCl2, 11 EGTA, Aspartate 10 HEPES (pH = 7.2 titrated with KOH). The extracellular (bath) solution contained (in mM) 115 NaCl, 20 TEA-OH, 1 MgCl2, 2 CaCl2, 5 glucose, 10 HEPES (pH = 7.4 titrated with NaOH), supplemented with 600 nM TTX when recording rat or human NaV1.8 channel currents. All channels were activated

using the following stimulation protocol: Holding level −100 mV, ramp from −100 to +60 mV (50 ms), delivered every 10 s and recorded at a sampling rate of 10–50 kHz. The ramp protocol is increasingly in use as a quick measure of I–V relationship (see for example Dib-Hajj et al., 2007). Indeed, it is possible that measuring inhibition upon square pulse stimulation, may have yield somewhat different results (maybe shifting the dose response curves). However, the ramp stimulation method has enabled the use of exactly the same stimulation protocol with all channels tested. All chemicals were from Sigma–Aldrich (Rehovot, Israel) apart from TTX from Alomone Labs (Jerusalem, Israel). All results are presented as mean ± standard deviation.

However, such post-synaptic effects are short-lived, so this expl

However, such post-synaptic effects are short-lived, so this explanation would require that CVS produces prolonged firing in vestibular afferents, and thus prolonged excitatory or inhibitory influence on bimodal neurons, throughout the time course of our experiment. An alternative explanation would involve a longer-lasting effect of the transient stimulation of vestibular peripheral organs on the cortical

targets of somatosensory pathways. Such enduring interactions are suggested by the lack of reduction of the modulatory effect observed across our five blocks of testing. CVS might perhaps produce long-lasting modulation of somatosensory synaptic strength by long term potentiation (LTP) of tactile pathways, and long term depression (LTD) of pain pathways. Further research is necessary to investigate these possible mechanisms of vestibular-somatosensory Cobimetinib concentration interaction. What could be the adaptive function of these vestibular modulations

of touch and pain? CVS is a very unnatural stimulus, so we can only speculate on this point. Outside the laboratory, vestibular canal input normally occurs during head rotation, as when an animal re-orients towards a new part of the external environment (Klam and Graf, 2006). We suggest that such reorienting may involve a rebalancing of sensory processing to provide an appropriate Pifithrin-�� in vitro new balance of inputs. For example, pickup of information from novel environments may become urgently important following reorienting (Fecteau et al., 2004). Thus, vestibular signalling of head rotation during orienting movements could trigger increased sensitivity to tactile stimuli. Interestingly,

our data suggest that vestibular input causes a complementary tweaking of the sensitivity of the two main submodalities of somatosensation, buy C59 rather than a general reduction or increase in sensitivity of them. Interestingly, the observation that vestibular input has an analgesic effect is reminiscent of the notion that novel environments are themselves mildly analgesic (Siegfried et al., 1987). The observed tweaking of the sensitivity of the two somatosensory submodalities may reflect a multisensory mechanism for adjusting sensory processing following reorientation to novel environments, thus ensuring efficient perception and motivating exploratory behaviour (Cohen et al., 2007). This work was supported by EU FP7 project VERE and by a Leverhulme Trust Major Research Fellowship to P.H., E.R.F. was supported by a PhD program of the University of Pavia, and by a BIAL Foundation Bursary (215/10) awarded to PH. G.B was supported by PRIN 2007. G.D.I. is University Research Fellow of The Royal Society and is supported by the BBSRC and El.En. “
“Luigi A. Vignolo, M.D. passed away peacefully at home, surrounded by his family, on December 21st, 2011.

Although it may be associated with any kind of neoplasm, TS is mo

Although it may be associated with any kind of neoplasm, TS is most often related to pancreatic, lung, prostate, gastric, colorectal, ovarian and breast cancer.9 A 58-year-old man, electronics technician, was admitted in our Internal Medicine ward with deep venous thrombosis of the right lower limb. He presented Z-VAD-FMK price to the Emergency Department with a 3-day course of right calf pain worsened by walking, followed by swelling and increased temperature in the same limb. Throughout the whole period he felt increasing fatigue and had an episode of fainting. Just four days before the current symptoms started he had arrived from a vacation in Ecuador, during which his right upper limb had become

swollen, red and hot. He was diagnosed with right arm cellulitis and was started on antibiotic and anti-inflammatory therapy, improving subsequently. He denied fever, sweating, weight loss or coughing, as well as any digestive, urinary or other musculoskeletal symptoms. Past medical history was positive for some

childhood infectious diseases (measles, mumps, chicken pox), grade I arterial hypertension (known for 21 years and without medication), smoking habits (20 pack-year units), mild alcohol intake (20 g daily), chronic lumbar disc disease, left varicocele surgery (at the age of 21) and benign prostatic hypertrophy. His father deceased, with a history of chronic renal failure. There were no discernible accounts of cancer in close relatives. His physical examination revealed great overall condition and stable vital signs (BP 113/70 mmHg, HR 70 bpm, RR 20 bpm, apyrexia); no skin lesions, lymphadenopathy or thyromegaly; normal cardiac and respiratory sounds; soft, nontender, nondistended abdomen with normal bowel sounds, no masses on abdominal examination, and no hepatosplenomegaly; no evidence of infection in his right upper limb; slight swelling and increased temperature in his right leg, with positive Homans’ sign; normal neurologic exam and fundus observation within normal limits. many Laboratory tests showed the following: haemoglobin 14.6 g/dl; WBC 10.9 × 109/l

(68.1%N–20.3%L–7.1%M–4%E); platelets 258.0 × 109/l; ESR 13 mm; CRP 3.5 mg/dl (N < 1); transferrin 195 mg/dl (N: 215–365); ferritin 344.9 ng/ml (26.0–388.0); glucose 84 mg/dl; creatinine 0.6 mg/dl; albumin 3.7 g/dl; normal serum electrophoresis; AST 42 U/l (17–59); ALT 65 U/l (21–72); GGT 168 U/l (N: 15–73); ALP 209 U/l (N: 38–126); total bilirubin 0.4 mg/dl; amylase 591 U/l (N: 30–110); lipase 6356 U/l (N: 23–300); LDH 704 U/l (N: 303–618); total cholesterol 180 mg/dl; triglycerides 122 mg/dl; total calcium 9.5 mg/dl; INR 1.1; aPTT 38.0′′; factor V 130.5%; factor VIII 152.2%; protein C 97%; protein S 92.8%; antithrombin III 107%; resistance to activated protein C 3.14 (within normal limits); Lupus anticoagulant 1.94 ratio (1.6–2.0), Silica clotting time 1.26 ratio (>1.