The bone marrow cells were placed in duplicate 1 mL semisolid agar cultures in 35 mm Petri dishes using 1 × 105 bone marrow cells per culture for the growth of CFU-GM. The cells were cultured in Dulbecco’s Modified Eagle’s Medium (DMEM, Sigma Chemical Co., St. Louis, MO) containing 20% FCS (fetal calf serum) and 0.3% agar. Colony formation was stimulated by the addition of recombinant murine macrophage–granulocyte colony-stimulating factor (rmGM-CSF-Sigma) at a final concentration Obeticholic Acid in vitro of 0.5 ng/mL. The

cultures were incubated for 7 days in a fully humidified atmosphere of 5% CO2 in air, and colony formation (clones >50 cells) was scored at 35× magnification using a dissection microscope (Metcalf, 1984). To evaluate the hematopoietic cell populations, whole BM and LTBMC cells were collected by flushing (1 × 106 cells), fixed and labeled. To the verification of mature cells we used 4 antibodies conjugated with four different

fluorocromes: FL1: anti-Gr1-FITC; FL2: anti B220-PE, FL-3:anti-Mac-1-Cy7/PE and FL4: anti-CD3-APC. To analyze the primitive population we used 2 antibodies that Rucaparib ic50 recognize the fraction LSK together with a cocktail of mature lineage: FL2: anti-B220, anti-CD3, anti-Ter-119, anti-CD11b and anti-Gr-1, which were all conjugated with PE; FL3: anti Sca-1-Cy7/PE and FL4: anti-c-kit-APC. The data were collected using a FACSCalibur flow cytometer and analyzed using CellQuest software (BD Biosciences). The antibodies were purchased from BD Biosciences. The mice were bled from the heart under deep halothane anesthesia. Within each experimental group, the blood was pooled, left at 37 °C for 30 min, and the clots were allowed to retract overnight at 4 °C. Following centrifugation, the serum was removed and stored at −20 °C. CSA was determined

by measuring the ability of serum obtained from control and experimental groups to www.selleck.co.jp/products/Rapamycin.html stimulate HP to form CFU-GM (1 × 105 cells) from normal mice. The results were expressed as units of CSA/mL, where 1 unit/mL was defined as the lowest amount of CSA able to induce the formation of colonies (Van Den Engh and Bol, 1975). Marrow cells were aseptically collected from two complete femur shafts after killing the animal by cervical dislocation. The plug of marrow cells was gently extruded into a sterile plastic tube using 1 mL of RPMI 1640 medium (Sigma) injected through the femur and then converted to a dispersed cell suspension in 5 mL of RPMI by gently aspirating the suspension up and down 20 times using a sterile 5 mL pipette. To establish the culture, 1 × 107 pooled femoral bone marrow cells were dispensed into T25 tissue culture flasks containing 10 mL of RPMI 1640 supplemented with 25 mM l-glutamine, 25 mM HEPES, 200 UI/mL penicillin, 100 μg/mL streptomycin, 20% horse serum (Sigma), and 0.1 μM hydrocortisone and incubated at 37 °C in 5% CO2.

No association with the diagnosis of major depressive episode during the course of IFN-α therapy was observed genotype or allele-wise (p > 0.05). Multivariate logistic regression analyses including fibrosis, current major depression, current anxiety disorder,

report of psychiatric treatment, current suicide risk, current BDI and HADS scores, as well as the genotype groups and genetic ancestry estimations, confirmed the lack of association NVP-BEZ235 mouse between the rs10089084 (OR = 1.17; 95% CI = 0.56–2.45; p = 0.676) and the rs3824259 polymorphisms (OR = 1.10; 95% CI = 0.50–2.41; p = 0.810), and the diagnosis of IFN-α-related depression. The enzyme IDO is known to act by metabolizing tryptophan in serotonin and kynurenine. Although the role of IDO in IFN-α-induced depression is supported by many

studies (Wichers and Maes, 2002, Bonaccorso et al., 2002, Capuron and Miller, 2004 and Comai et al., 2011), to the best of our knowledge, this is the first study to investigate the influence of the genetic variants of this enzyme and the diagnosis of major depression during the course of IFN-α therapy. Contrary to our hypothesis, no association between the rs3824259, rs10089084, and rs35099072 polymorphisms and IFN-α-related depression was indentified. We accounted for the potential bias related to population stratification Cabozantinib concentration in the Brazilian ethnically heterogeneous population using thirty-five AIMs that show large differences in allele distribution among the three main ethnic groups (European, African and Indigenous). However, the inclusion of the estimated ancestry did not affect the genetic association results.

It is important to note that the high level of admixture found in our sample has a strong influence on the haplotype structure of the gene, and therefore other SNPs in and around the gene should be further evaluated before any conclusion regarding the effect of the IDO gene variation in the predisposition of IFN-α-related depression can be reached. In addition, power calculation revealed that the total sample has approximately 80% power to detect differences in genotype group frequency > 18%, assuming the frequency of 46.8% and 63.5% of the CG/GG and GT/GG genotype groups among individuals who have not developed IFN-α-related depression, for the rs10089084 and rs3824259, respectively. The fact that Methocarbamol an association between these polymorphisms and the diagnosis of depression related to IFN-α therapy was not found in our HCV patients suggests that other genetic variations either influence or are influenced by IDO and its metabolites’ actions. Indeed, polymorphisms of pro-inflammatory cytokines that may be associated to the overstimulation of IDO, such as IL-6 (Bull et al., 2009) and IL-28B (Lotrich et al., 2010), of the IFN-α-receptor 1 (Yoshida et al., 2005), the serotonin transporter (5-HTT) (Bull et al., 2009 and Lotrich et al., 2009), and the serotonin-1A receptor gene (HTR1A) (Kraus et al.

The tanks are structurally complex and composed of interconnected bays, longitudinal and transverse stringers/stiffeners to improve the strength of the vessel. The usual layout of ballast tanks on a bulk carrier consists of the tanks located at the fore peak, aft peak,

upper/topside wing, lower/hopper wing and bottom. The double bottom tank and hopper tank are unified and in some cases are connected with the upper wing/topside tanks by a trunk that allows the ballast water to flow between them. Fig. 1 shows a schematic of the ballast tanks of a bulk carrier. Other tankers have slimmer ballast water tanks along the ship and do not alternate. These ballast tanks are large with a simple box design, and have a capacity of 40,500 m3 PF-562271 of water serviced by pumps with a flow rate of 3000 m3/h (or ~1 m3/s). Inside the double bottom tank, individual compartments are generated by crossing longitudinal and transverse stiffeners and frames with lightening holes. The Selleckchem Dabrafenib neighbouring compartments are associated with lightening holes, stringers and limber holes, shown in Fig. 2. The ballast tank flushing is achieved either from the inlet as shown in Fig. 1(b) by the sequential (empty/refill) method or

through overflow arrangements by the flow through method. For the flow-through method, the overflow is achieved from two air/sounding pipes either on the deck or to the side, typically with a diameter of 0.15–0.2 m. The NIS that can be drawn into a ballast tank range from bacteria, plankton, fish eggs or crabs to fish (see Wonham and Carlton, 2005). Associated with these is a settling or swimming velocity, ranging from 0.1 to 150 mm/s (see Wong and Piedrahita, 2000 and Magill et

al., 2006). The smaller species are essentially advected with the flow and can be regarded as essentially passive during flushing. When the species are passive, the fraction of the original water that is flushed out of the ballast tank can be used as a proxy for estimating the removal of NIS from the tanks. The current legislation deals with the number of exchange volumes that are required to achieve a level of flushing. Future treatment strategies are likely to do with reflushing and cleaning while the click here ship is in transit, and again, knowledge of the distribution of treated ballast water will be useful. There are comparatively few theoretical studies of the flow within multi-compartment tanks. Wilson et al. (2006) and Chang et al. (2009) used CFD to examine the movement of fluid in a 1/3-scale double bottom tank and a full-scale ballast tank from a typical bulk carrier. When density contrast between the incoming seawater and the original freshwater was relatively large, the predicted flushing efficiency fell short of the required 95% replacement after three volumes exchange for both tanks, due to trappage in the tank tops.

2). To overcome the inherent problems and establish the clinical significance

of transcranial ultrasound perfusion imaging, we have clinically introduced the Sonopod for TCDS monitoring [10] and [11] and evaluated acetazolamide (ACZ) vasoreactivity [12] and [13]. The objective of this study is to clarify clinical usefulness and identify problems in TCDS-Sonopod monitoring in the evaluation of brain tissue perfusion. Brain tissue perfusion monitoring was evaluated in 11 patients (ages 31–94, mean 66). Details of patient demographics are shown in Table 1. After a 5 ml-bolus Levovist® injection (2.5 g, 400 mg/ml) via the antecubital vein, power modulation imaging (PMI) in all cases in comparison with second harmonic imaging (SHI) in the initial two cases were evaluated in the supine position via TWs NU7441 in vitro (Fig. 3). Both imaging types were visualized by an integrated backscatter method. The transmitting and receiving frequencies Cobimetinib chemical structure of PMI and SHI were 1.7/1.7 MHz and 1.3/2.6 MHz, respectively. The investigation depth was 16 cm with a focus of 8 cm. Settings were mechanical index 1.6, system gain 75, and compression 70. ACZ cerebral vasoreactivity, before and after 500 mg Diamox® intravenous injection,

was evaluated in 10 cases utilizing a SONOS5500 S3 transducer (Philips Electronics Japan, Ltd.) installed in the Sonopod. Time–intensity curves (TICs) on the diencephalic horizontal crotamiton plain were evaluated

before and after ACZ in five regions of interest (ROI); bilateral basal ganglia (BG) and thalamus (Th), and contra-lateral temporal lobe (TL). A total of 30 TICs with a duration of 10 min via the bilateral (five cases) and unilateral (five cases) TWs were analyzed before and after ACZ. Conventional SHI and PMI utilizing hand-held monitoring were compared in two cases. In the visualization of the contralateral hemispheres via the unilateral TWs, PMI was superior to SHI as shown in the upper panels of Fig. 4a and b. As shown in the lower panels of the quantitative TIC evaluations in both PMI and SHI, peak intensity (PI) in the contralateral hemisphere ROIs was lower than in the ipsilateral hemisphere ROIs. During hand-held monitoring, TICs were not always stable in all cases and drifted from the base-line due to patients’ movements as shown in the lower panels of Fig. 4. All patients could be fitted and monitored continuously by one examiner. Brain tissue perfusion could be precisely quantified before/after ACZ in the same ROI as shown in Fig. 5. Due mainly to patient’s movements, drifts from the base-line were observed in the TICs of 4 (seven TIC analyses) out of 10 (30 TIC analyses) patients. However, fixed-probe shifts due to patients’ movements during monitoring were easily re-adjustable and the TICs could be returned to the baseline in all patients as shown in Fig. 6.

The

SLR algorithm is based on relating target magnetization profiles (Mx,MyMx,My, and MzMz) to spinor parameter profiles (αα and ββ) whose discrete Fourier transform (DFT) coefficients can be inverted to obtain the RF pulse that produces them. To apply the algorithm to design an ΔωRF(t)ΔωRF(t) waveform that excites a slice along the |B1+| axis, we must express target excitation profiles in terms of the rotated αα and ββ parameters. The inverse SLR transform can then compute the ΔωRF(t)ΔωRF(t) waveform that corresponds to those parameters. Given initial magnetization Mzy-≜Mz-+ıMy-, and Mx-, the magnetization after a pulse with rotated αα and ββ parameters will be: equation(2) Mzy+Mzy+∗Mx+=(α∗)2-β22α∗β-(β∗)2α22αβ∗-α∗β∗-αβαα∗-ββ∗Mzy-Mzy-∗Mx-. For initial magnetization at thermal equilibrium ( (Mx-,My-,Mz-)=(0,0,1)), the excited TGF-beta inhibitor transverse magnetization will be: equation(3) Mx+=-α∗β∗-αβ=-2αRβR-αIβI equation(4) My+=I(α∗)2-β2=-2αRαI+βRβI,where the R   and I   subscripts denote GDC-0068 the real and imaginary parts of the parameters, respectively. As in conventional linear-phase SLR pulse design and previous |B1+|-selective design methods, we will design pulses that produce constant-(specifically, zero-) phase profiles across the excited slice so that My+=0. For these pulses βIβI will also be zero. If we further restrict our consideration

to small-tip-angle pulses with A(t)A(t) waveforms that have zero integrated area, then αR≈1αR≈1 and αI≈0αI≈0 [18]. In this case, equation(5) Mx+=-2βR,and My+=0. Therefore, βRβR is the parameter P-type ATPase of interest for digital filter design in the |B1+|-selective SLR algorithm. Conveniently, because Mx+=-2βR also for a conventional refocused small-tip-angle slice-selective pulse [18], the same ripple relationships provided in Ref. [16] also apply to |B1+|-selective pulse design. Fig. 2 illustrates the target ββ profile configuration. Unlike conventional slice-selective excitation, a |B1+|-selective slice profile cannot be centered at |B1+|=0, since excitation cannot occur with

zero RF field. Thus, the slice profile must be shifted away from this point. A slice-selective excitation is conventionally shifted using frequency modulation of the RF pulse; however, this would result in complex ββ DFT coefficients, and subsequently a complex-valued ΔωRF(t)ΔωRF(t) waveform. The ΔωRF(t)ΔωRF(t) waveform must be real-valued to be physically realizable, which dictates that the ββ DFT coefficients must be purely imaginary, since a small-tip RF pulse designed by SLR is π/2π/2 out of phase with its ββ DFT coefficients [16]. The required purely imaginary ββ DFT coefficients can be obtained by specifying an odd and dual-band (anti-symmetric) ββ profile [19]. Thus, the target ββ profile must be real-valued, dual-band, odd, and zero at |B1+|=0. The corresponding ΔωRF(t)ΔωRF(t) will be real-valued and odd. A real-valued, odd, and dual-band ββ profile and its corresponding DFT coefficients can be designed in several ways.

“
“Extreme weather events have severe consequences for human society. The impacts of the changing climate will likely be perceived most strongly through changes in intensity and frequency of climate extremes. Studies have found that human activities have contributed Daporinad to an increase in concentrations of atmospheric greenhouse gases contributing to intensification of heavy rainfall events (Min et al., 2011). In the context of hydrology, the changing climate will likely accelerate the hydrological cycle on a global scale, and subsequently intensify the uneven spatial and temporal distribution of hydrological

resources (Huntington, 2006 and Trenberth, 1999). The intensity of extreme rainfall events is projected to increase under global warming in many parts of the world, even in the regions where mean rainfall decreases (e.g., Semenov and Bengtsson, 2002 and Wilby and Wigley, 2002). Thus climate adaptation strategies for e.g. emergency planning, design of engineering structures, reservoir management, pollution control, or risk calculations rely on knowledge of the frequency of these extreme events (Kumke, 2001). Assessment of these extreme rainfall events is important in hydrological DZNeP mouse risk analysis and design of urban infrastructures.

The increasing trend of rainfall extremes has quantifiable impacts on intensity duration frequency relations (Kao and Ganguly, 2011), and an increase in the intensity and/or frequency of extreme rainfall events ioxilan may result in the flooding of urban areas (Ashley et al., 2005 and Mailhot et al., 2007). In India, rainfall variability is a central driver of the national economy as it is predominantly agricultural. A change in extreme events would have a large impact on the growing economy of India as most of the population live in urban areas. Several studies have

addressed the issue of trends in rainfall in India since last century. Long-term southwest monsoon/annual rainfall trends over India as a whole were previously studied by Parthasarathy et al. (1993) and Rana et al. (2012), among others. Long term trends for the last 50 years indicate a significant decrease in the frequency of moderate-to-heavy rainfall events over most parts of India e.g., Dash et al. (2009) and Naidu et al. (1999). This is corroborated by a significant rise in the frequency and duration of monsoon breaks over India during recent decades (Ramesh Kumar et al., 2009 and Turner and Hannachi, 2010), while the frequency of extreme rainfall events (100 mm/day) have increased in certain parts of the country (Goswami et al., 2006). Future climate studies for India based on climate model simulations suggest that greenhouse driven global warming is likely to intensify the monsoon rainfall over a broad region encompassing South Asia (e.g., Lal et al., 2000, May, 2002, May, 2004, May, 2011, Meehl and Arblaster, 2003 and Rupakumar et al., 2006).

The width of the stenotic canal can often be

measured in higher degrees of stenosis as well with B-mode imaging. The diameter can then be related to the distal one for measuring the degree of stenosis following the NASCET method, but this is only possible with excellent conditions for insonation. Color Doppler is helpful in delineating plaques of low echogenicity or proving BMN 673 chemical structure absence of flow in the occluded ICA. But it does not allow precise diameter measurements due to its low frame rate and a huge influence of the gain. Grading of stenoses above 50% is the basis of clinical decisions. Combining morphologic and several hemodynamic features allows a reliable description of at least four classes of stenosis. Such a multiparametric approach avoids severe misclassification as is done with a simplified PSV criterion or its derivates alone (end diastolic velocities in the stenosis, ratio of velocities ICA/CCA). Secondary criteria may be helpful in supporting the diagnosis as the extend of flow disturbances being most pronounced in a 70–80% stenosis and diminishing selleck chemical together with a reduced flow volume in very a high degree stenosis In a high degree stenosis the hemodynamic effect is shown by the appearance of collateral flow, which is driven by the poststenotic pressure drop. Another effect is a poststenotic decrease of velocity and pulsatility of flow. All these effects can be measured reliably by extra-

and intracranial Doppler duplex sonography. The question is whether the trial result that surgery is highly beneficial in case of a symptomatic ≥70% NASCET stenosis as measured by angiography can be translated into: beneficial in case of a “hemodynamically relevant stenosis” because 70% stenosis is the threshold from which a pressure drop and decreased poststenotic flow can be observed. This seems reasonable but is so far not accepted as level

one evidence. [8]. A meta-analysis of studies correlating PSV and percent of stenosis as measured by angiography showing a considerable disagreement was the background of not accepting ultrasonography. The old concept of a multiparametric diagnosis was not considered. However it has been used and taught over decades. New technical elements have been continuously introduced. But there Phosphoprotein phosphatase is a lack of well designed and large studies for this concept, including all these new techniques. In older publications e.g. the definition for measuring the degree of stenosis (NASCET or ECST) is missing. This is one of the reasons why, they do not add very much to the evidence. Even with such new studies some disagreement between methods will persist as explained above. Clinically most useful would be to repeat randomized carotid surgery trials with ultrasonography as criterion for decision in symptomatic patients. However it is ethically not justified to randomize for this question again.

In pre-mRNA processing the multi-domain splicing factor U2AF65 recognizes a uridine rich RNA sequence to promote spliceosome assembly. The protein possesses two RNA recognition motifs, RRM1 and RRM2 connected by a flexible linker. PREs data obtained by spin-labelling different residues of either RRM1 or RRM2 in the RRM1–RRM2 construct revealed the presence of a conformational equilibrium between PD-0332991 cost an “open-state”, where both RRM domains are capable of binding the RNA, and a “closed-state”, where only RRM2 binds to the RNA and the RNA binding surface of RRM1 is partially engaged

in electrostatic interactions with RRM2. By analysing the percentage of “open” versus “closed” conformations in the presence of substrate RNAs of different sequence, the authors could correlate the amount of protein in the “open-state” with the efficiency GSK1120212 of the U2AF65–RNA interaction in promoting spliceosome assembly. Furthermore, they could demonstrate

that protein mutations destabilizing the “open-state” are impaired in their ability to bind the RNA. This study demonstrates the usefulness of PRE data for characterizing the relative orientation of protein domains or of distinct components of a complex, including even the detection of multiple conformations. An extensive set of PRE-derived distances can be used to guide molecular docking and determine the conformation of RNP complexes. As mentioned above, site-directed paramagnetic labelling of proteins is only possible in the absence of multiple accessible cysteines. If more than one cysteine is located

on the surface Parvulin of the protein, these residues can be mutated to serine, under the provision that mutagenesis does not alter the protein folding. Alternatively, a different implementation of the PRE effect has been proposed, which does not requires site-directed spin-labelling [44]. A soluble paramagnetic agent Gd(DTPA–BMA) (DTPA: diethylenetriamine pentaacetic acid, BMA: bismethylamide) is added to the solvent, resulting in line broadening of the accessible nuclear spins. This data can be translated into structural information defining the distance of the nuclear spins from the surface, or in other words the solvent accessibility (Fig. 4). Solvent PREs have been used in a combined structure-selection/structure-refinement protocol to calculate the conformation of the Ran-CRM1-PKI NES complex together with sparse NOEs [44]. More recently an empirical function translating solvent accessibility data into structural information has been implemented in Xplor-NIH for structure calculations [45]. A similar approach has been applied to nucleic acids as well.

Two different hSNCA-expressing control groups were used in this study, one that received AAV-hSNCA alone and one that received AAV-hSNCA and a control, non-silencing, silencing vector containing a scrambled sequence (AAV-NS), which interestingly differed in some of the toxic effects examined. Both hSNCA-expressing groups exhibited a similar forelimb motor deficit and similar loss of TH-IR neurons in the SN by 1 month. However, rats that received AAV-hSNCA and AAV-NS exhibited greater toxic effects than those observed in rats that received AAV-hSNCA alone, which included selleck chemicals loss of TH-IR

fibers in the ST, reduction in total TH expression in the ventral midbrain and the ST, as measured by western blot, and an increased inflammatory response as detected by Iba-1-IR. The greater toxicity observed in rats treated with AAV-hSNCA and AAV-NS could be attributed to silencing vector design, off-target effects of the scrambled sequence, Mitomycin C nmr or to increased

viral load. It is also possible that co-injection of silencing vector resulted in some unknown modulatory effect on hSNCA vector expression. Rats that received hSNCA alone did exhibit some reduction in total TH expression in the ventral midbrain and ST, as measured by western blot, although this reduction was not significant. A lack of toxicity on TH-IR fibers in the ST by AAV2/8-hSNCA alone, has also been observed in some studies where hSNCA was delivered to the SN using either AAV2/6 (Azeredo da Silveira et al., 2009) or AAV2/8 (McFarland et al., 2009). However, other studies using AAV2/2, 2/5 or 2/6 have shown hSNCA-induced reductions in TH-IR fibers in the ST by 8 weeks (Decressac et Progesterone al., 2012, Gorbatyuk et al., 2008, Kirik et al., 2002 and Lundblad

et al., 2012). These differences most likely reflect varying levels of hSNCA protein in DA axons due to differences in vector dose, serotype and/or efficiency of retrograde transport, but may also result from toxic effects at different ST levels since only one ST level was quantified. The AAV-hSNCA and AAV-NS group is the most appropriate hSNCA-expressing control group for assessment of effects due to hSNCA gene silencing with mir30-SNCA because both of these groups received similar viral load and were injected with similar vector constructs. When compared to the AAV-hSNCA and AAV-NS group, hSNCA gene silencing with mir30-SNCA results in significant protective effects on forelimb motor behavior, TH-IR neurons in the SN and TH-IR fibers in the ST. However, toxic effects that may have resulted from high viral load or from silencing vector design were observed in both the NS and mir30-SNCA groups.

This impaired mineralisation has been shown to alter the bone material quality and the functional biomechanics of the tissue at micro- [14] and nanoscale levels [15]. In this study, we have used an N-ethyl-N-nitrosourea (ENU) induced mouse mutant for X-linked hypophosphatemic rickets

(Hpr), arising from a Trp314Arg missense mutation in the Phex (phosphate-regulating gene with homologies to endopeptidases on the X-chromosome) gene [15], and focused our studies on the scapula for the following reasons. The scapula is a large triangular flat bone which has five thick bony ridges (glenoid, scapula spine, medial and lateral border click here (LB) and caracoid process) and two hard flat bony structures, denoted as infraspinous fossa (IF) and supraspinous fossa [5]. The scapula is subject to a number of muscle, ligament and joint reaction forces during movement, and the location, magnitude and direction of these forces differ extensively between tissue regions within the same scapula. Indeed, the force variation at different muscle insertion points

can be very large, Selleckchem Ruxolitinib with a spatially variable stress distribution ranging from 0.05 MPa at IF versus 60 MPa at LB estimated using finite element modelling at the macroscale [5]. We therefore utilised the scapula from Hpr mice as a model system to investigate muscle force-mediated mineral particle orientation and its alteration

due to defective bone mineralisation, using synchrotron scanning X-ray nanoimaging methods. Advances in synchrotron X-ray sources generate X-ray beams of micrometre size (1–10 micrometres), Teicoplanin allowing scanning SAXS experiments to map spatial variations in the nanostructure with a high resolution [16]. This technology enables quantitative investigation of the nanocrystallite organisation in the tissue, with micron-scale resolution. An ENU induced mouse model for X-linked hypophosphatemic rickets (Hpr) arising from a missense Trp314Arg Phex mutation was used [15]. Wild-type and Hpr male mice aged 1, 4, 7 and 10 weeks were studied. Mice were kept in accordance with UK Home Office welfare guidelines and project licence regulations. Dissected scapulae from 1, 4, 7 and 10 week old mice were skinned, cleaned of muscle tissue, wrapped in gauze, soaked in phosphate buffered saline (PBS), and stored at − 20 °C until the scanning SAXS experiment was conducted (approximately 1 week). Just before the experiment, each scapula was mounted in a saline sample chamber with Ultralene® (SPEX SamplePrep, Metuchen, NJ, USA) foil windows, as shown in Fig. 1(A). For the scanning SAXS measurements of specimens, 3.4 mm2 (Fig. 1(B)) areas were selected.