Dieser Befund bildete die Grundlage für die Hypothese, dass MeHg

Dieser Befund bildete die Grundlage für die Hypothese, dass MeHg in den Gliazellen demethyliert und anschließend BIRB 796 das entstandene Quecksilber in die Neuronen transportiert wird, wo es seine Neurotoxizität entfaltet. Auf diese Weise könnte die „Auswahl” der Neuronen, die geschädigt werden, in den benachbarten Gliazellen erfolgen, wo die Demethylierung von MeHg abläuft. Das späte Einsetzen der Symptome ließe sich durch den langsamen Prozess der Demethylierung und des Transfers des Quecksilbers aus den Gliazellen in die Neuronen erklären. Magos und Clarkson [106] stellten eine Methode vor, mit der das Gesamt- und das anorganische Quecksilber in derselben

biologischen Probe ermittelt

werden kann. Mithilfe dieser Methode bestimmte Syversen [107] den Gesamtquecksilbergehalt sowie den Gehalt an Hg2+ in subzellulären Fraktionen von Rattenhirnen MG-132 solubility dmso nach einer einzelnen intravenösen Injektion von 203Hg-markiertem MeHgCl oder HgCl2. Die Daten zeigten, dass der Hg2+-Gehalt im Gehirn nach Injektion von MeHg etwa 20-mal höher war als nach Injektion einer ähnlichen Dosis von HgCl2. Dies unterstreicht, dass im Hirngewebe Demethylierung stattfindet und dass durch diesen Prozess mehr intrazelluläres Hg2+ produziert wird, als über die Blut-Hirn-Schranke aufgenommen wird. Die Hg2+-Spitzenkonzentration im Gehirn wurde einen Tag nach HgCl2-Exposition, jedoch erst 8 Tage nach MeHg-Exposition erreicht. Garman et al. [108] verabreichten Makaken 203Hg-markiertes MeHg über eine Magensonde

und untersuchten deren Gehirne mittels Histopathologie und Autoradiographie. Die Autoradiographien wurden erstellt, indem Gewebeschnitte mit einer photographischen Silberhalogenidemulsion behandelt wurden. In einer Notiz am Ende des Artikels wird jedoch erwähnt, dass die Autoradiogramme nicht das Isotop, sondern den Hg-Ag-Komplex zeigen, der in der Emulsion entsteht. Solch ein Komplex kann sich nur zwischen Hg2+ und Ag ausbilden, nicht zwischen MeHg und Ag. Der Großteil der Radioaktivität (die Hg2+ repräsentiert) lag in den Gliazellen check vor und nicht in den Neuronen. Sakai et al. [109] führten eine ähnliche Silbermarkierung an Gehirnschnitten von Minamata-Opfern durch und zeigten, dass sich der Großteil der Radioaktivität in den Gliazellen befand, obwohl auch in den meisten anderen cerebellären Neuronen Hg-Ag-Körner zu sehen waren. Einmal mehr sollte betont werden, dass diese anorganisches Quecksilber und nicht das Gesamtquecksilber repräsentieren. Magos et al. [56] verglichen die Neurotoxizität von MeHg und Ethylquecksilber. Nach Exposition gegenüber Ethylquecksilber im Vergleich zu MeHg war die Hg2+-Konzentration höher, wobei eine Schädigung der Körnerzellschicht nur nach MeHg-Exposition erkennbar war.

We suppose

that this fact might be due to inhaled adminis

We suppose

that this fact might be due to inhaled administration of the corticosteroids having a topic anti-inflammatory effect, promoting lower systemic concentrations of the drug. Even if some adverse effects were expected by the corticosteroid administration, they are less pronounced when compared to other ways such as injections or via gastric administration.27 This is the specific reason why we used topical administration. Additionally, the specific research hypothesis was related to the possible adverse effects in the periodontium when corticosteroids are inhaled such as in asthma treatments, for example. There is an increase production of TNF-α in G2 as compared to G1. This finding was in agreement with the literature.28 This was probably due to the presence of periodontal inflammation, which reinforces that UK-371804 order periodontal disease is a systemic problem.29 and 30 No effect Alectinib molecular weight in TNF-α production was observed when G3 and G4 are analysed. This also strengthens the idea that inhaled steroids have little systemic anti-inflammatory effect.31 On the other hand, the literature has shown that budesonide has a high level of topical anti-inflammatory activity.32 Nevertheless, this topical glucocorticoid was not able to modulate the periodontal breakdown. The results of the present study can contribute for better understanding of the pathogenesis of alveolar bone loss. Data suggest that inhaled corticosteroid does not significantly

interfere in the mechanisms of bone loss. This can be interpreted that budesonide does not have a promising effect as a modulator of host response to future studies such Fenbendazole as other anti-inflammatory drugs.7, 33 and 34 Additional studies evaluating effects of budesonide in oral mucosa are encouraged. We concluded that different concentrations of inhaled budesonide do not interfere in the ligature-induced alveolar bone loss in Wistar rats. Additionally, no effect of budesonide was observed in TNF-α production. We would like to thank Lorena F. Orlandini for helping us with the animals. Funding: The present study was partially funded by PROCAD (Academic Cooperation Project)

Grant number NF2341 (08/2008). Competing interest: None declared. Ethical approval: The research protocol was approved (protocol number 2008128, Sep 24 2009) by the Ethical and Research Committee of the Federal University of Rio Grande do Sul. “
“The lateral parabrachial nucleus (LPBN), a pontine structure located dorsolaterally to the superior cerebellar peduncle (SCP), is an important hindbrain area involved in the inhibitory control of ingestive behaviour.1 and 2 The LPBN might also regulate central responses produced by systemic immune stimuli.3 and 4 It has been demonstrated that the LPBN plays a critical role in cytokine-induced Fos expression in the central nucleus of the amygdala (CeA), bed nucleus of the stria terminalis (BNST) and ventrolateral medulla (VLM) neurons.

The sample surface was carbon coated prior to qBEI A digital ele

The sample surface was carbon coated prior to qBEI. A digital electron microscope (DSM 962, Zeiss, Oberkochen, Germany) equipped with a four quadrant semiconductor BE detector was used for backscattered electron imaging. The accelerating voltage of the electron beam was adjusted to 20 kV, the probe current to 110 pA, and the working distance

to 15 mm. The digital backscattered (BE) images of trabecular bone areas were acquired by a single frame with a scan speed of 100 s/frame and a pixel resolution of 1 μm. Areas with high backscattered electron intensities – light gray levels – represent mineralized matrix with high Ca contents, whereas areas with low intensities – dark gray levels – indicate low mineral density. For the characterization and quantification of changes learn more in the bone mineralization density distribution (BMDD) curve, four outcomes were used: CaMean (the weight mean calcium content of the bone area obtained from the integrated area under the BMDD curve), CaPeak (the mode of calcium content indicated by the peak position in the BMDD diagram), CaWidth (the heterogeneity of mineralization

caused by the coexistence of BSU of different ages measured at the full width at one-half maximum of the BMDD-peak), CaLow (reflects the fraction low mineralized bone areas (< 17.68 wt Ca)). Details of the analysis method have been published previously [26]. Additionally, these qBEI images were also used later, to evaluate this website the mean gray level/mineral content (CaInd) at the nanoindentation sites similar as described in Ref. [27]. After μCT scanning, the L2 vertebral bodies were rehydrated, mounted in a servo-hydraulic testing system (858 Mini Bionix II, MTS, USA), preconditioned with 10 cycles in the elastic range and tested to failure Quisqualic acid in axial compression

with a rate of 0.033 mm/s. Stiffness, maximal load to failure, and energy to failure were computed from the resulting force–displacement curves. A mean tissue modulus for each vertebral body was then back-calculated from the experimental stiffness using a 12 μm resolution, homogeneous, linear elastic (v = 0.3) finite element model of the same loading scenario. Nanoindentation was performed using a Scanning Nanoindenter (Hysitron Inc., Minneapolis, USA) with a Berkovich diamond indenter tip as described elsewhere [28]. The calibration of the instrument was performed by doing indents of increasing depth in fused quartz with a known reduced modulus of 72 GPa. The material properties of the diamond tip are known such as the Poisson’s ratio is 0.07 and elastic modulus of the tip is 1140 GPa. Automated area scans of indents were performed using moving sample stage, which has a positional resolution of 1 μm. There was a distance of 10 to 11 μm between indents. A thermal drift correction factor was introduced automatically before each indent, by measuring the drift for 20 s.

However, we cannot exclude the possible presence of neurotransmit

However, we cannot exclude the possible presence of neurotransmitters or low molecular mass Everolimus nmr mediators in the S. plumieri venom, since they have been found in S. verrucosa and S. horrida stonefish venoms ( Garnier et al., 1996). The two-dimensional SDS-PAGE analyses showed that the majority of the S. plumieri venom components are in the mass range of 6–120 kDa and are predominantly

anionic proteins (pI 4–7). A similar MW range has been described for the protein components of other fish venoms: 20–295 kDa in Synanceja trachynis ( Hopkins and Hodgson, 1998), 11–109 kDa in Gymnapistes marmoratus ( Hopkins and Hodgson, 1998), 14–100 kDa in Thalassophryne maculosa ( Sosa-Rosales et al., 2005a), 15–130 kDa in Potamotrygon falkneri ( Haddad et al., 2004). Despite the fact that various proteins are found in the SpV, only the major spot observed in the two-dimensional electrophoretic profile of S. plumieri

venom was recognized by the SFAV after immunoblotting analysis. These in vitro observations correlate well with the results obtained in the in vivo assays and also corroborate that S. plumieri venom compounds responsible for inflammatory and cardiovascular effects are similar to those found Galunisertib research buy in stonefish venom. In addition, ELISA analysis of S. plumieri venom proteins suggested that the epitope(s) detected by the neutralizing polyclonal SFAV antibody is (are) shared by proteins present in both fish venoms. Interestingly, Andrich et al. (2010) demonstrated that SFAV was able to cross-react and neutralise the hemolytic activity of Sp-CTx, a dimeric (73 kDa/subunit) cytolytic and vasoactive glicoprotein isolated from S. plumieri venom ( Andrich et al., 2010). Metalloexopeptidase Thus, due to its MW

it is possible that the SFAV-recognized spot in the present work is the previously identified scorpionfish venom cytolysin. The isoeletric point variation of the SFAV-recognized protein spot could be due to the different glycosilation levels exhibited by Sp-CTx ( Andrich et al., 2010), being an additional evidence that the SFAV-recognized spot is the scorpionfish cytolysin. Both the molecular mass (98 kDa) and isoeletric point (6.0–7.0) values of SFAV-recognized protein spot are similar to the stonustoxin (SNTX; α subunit = 71 kDa, β subunit = 79 kDa, pI 6.9) and trachynilysin values (TLY; α subunit = 76 kDa, β subunit = 83 kDa, pI 5.7), the dimeric cytolytic toxins isolated from Synanceja horrida and S. trachynis venoms, respectively ( Poh et al., 1991, Kreger, 1991 and Colasante et al., 1996). The cytolysins from fish venoms are reported as multifunctional toxins, triggering an array of biological actions, including in vitro hemolysis, increase in vascular permeability, cardiovascular disorders and death ( Perriere et al., 1988, Poh et al.

Clear edge staining was also observed in P 1 PBECs, confirming th

Clear edge staining was also observed in P.1 PBECs, confirming the maintenance of BBB features after passaging. The loss of in vivo phenotype reported for many in vitro BBB models appears to be mainly due to the removal of endothelial cells from their natural STA-9090 research buy environment. However, the changes can be counteracted to some degree using several inductive factors and co-cultures as discussed. Recently developed primary cultured in vitro BBB models offer advantages as assay systems since they express more features of the in vivo BBB (including membrane lipid and protein composition, expression of uptake and efflux transporters and drug metabolising enzymes) than Caco-2

(from human colon carcinoma) or MDCK (from canine PD98059 clinical trial kidney epithelium) cell lines, which are commonly used in the pharmaceutical industry. Until around year 2000 the in vitro BBB model showing the best correlation with in vivo BBB permeability was the system using bovine brain endothelial cells co-cultured on filters above rat astrocytes

( Cecchelli et al., 1999), but over the last decade several groups have reported successful use of porcine brain endothelial cells as useful tools for drug screening ( Franke et al., 1999 Franke et al., 2000, Smith et al., 2007 and Zhang et al., 2006). Our results demonstrate that the PBEC model described here has the potential to be useful as a permeability screen to investigate BBB permeation of drugs of interest with a range of chemistries, including those that are substrates for transporters, whether or not the

particular transporters involved have been identified. With inclusion of sufficient passively permeating reference compounds, substrates for transporters can be identified as outliers, for further mechanistic study. If required and desirable, porcine brain endothelial cell production could be scaled ADP ribosylation factor up for high/medium-throughput screening. However, it is possible to limit the numbers of compounds that need to be tested on living BBB models using better in silico (computer-based) screens. Thus a serial and parallel screening process can be used to bring the numbers to manageable level (e.g. 200 cf. >100,000) for testing on an in vitro BBB model ( Abbott, 2004). In conclusion, results confirm that this optimised in vitro porcine BBB model is relatively simple to prepare, reliable and repeatable compared to most other static BBB models, and gives high TEER without the need for astrocyte co-culture. The quality, simplicity and robustness of the porcine BBB model make it an attractive model for industry to use in CNS drug discovery programmes and also for a variety of basic scientific projects. Because the method generates PBECs with high TEER, it is likely to show good apical: basal differentiation for other important BBB features, including receptors, transporters, enzymes and ion channels.

This may provide important information to inform interventions fo

This may provide important information to inform interventions for people with BJHS. In this study we investigated the muscle activity

Selleckchem ERK inhibitor within a hypermobile group compared to a healthy control group during postural and balance tasks. We hypothesised that BJHS leads to altered recruitment patterns in muscles of both the pelvis and the lower limbs. Subjects were recruited through email advertising within the Physiotherapy, Occupational Heath and Dietetics departments at the Imperial College Heathcare NHS Trust. Further recruitment involved email advertising within the author’s university year group and research colleagues. Ethical approval was obtained from the Imperial College Ethics Committee. Subjects were drawn from a larger study of individuals with and without knee osteoarthritis. The criteria to be included in the present study were healthy people aged between 18 and 50 years who had no clinical or radiological symptoms of knee osteoarthritis and who can walk without the use of an assistive device. The exclusion criteria were any neurological

or painful musculoskeletal conditions involving Copanlisib price the lower limbs, rheumatoid or any other systemic arthritis and obesity (Body Mass Index (BMI) >35). A total of 16 subjects (4 male and 12 female) were recruited with an average age of 28 years (range 22–45 years). Eight subjects (3 male) had BJHS and 8 subjects (1 male) were controls. Average height (SD), weight and BMI of the hypermobile vs control subjects were 1.6 (0.1) vs 1.7 (0.1) m, 64.8 (5.4) vs 68.6 (9.5) kg and 22.6 (1.4) vs 23.5 (3.7), respectively. There were no significant differences between groups for these parameters. Both hypermobile and control subjects were free from pain at the time of testing, and had no history of back pain. The Beighton Criteria (Beighton et al., 1973) was used to determine whether the subjects

were considered hypermobile. Subjects were shown the movements that make up the Beighton criteria and asked to reproduce them. One point was awarded for each of the nine movements that were re-produced. A score of 4 or greater was heptaminol considered hypermobile for the purpose of this study. Eight of the subjects were hypermobile with an average score of 7.4 (SD 1.7) and eight subjects were controls with an average score of 0.5 (SD 0.9). Seven of the 8 hypermobile subjects demonstrated lower limb hypermobilty (hyperextended knee joints); none of the control subjects had lower limb hypermobility. None of the subjects were seeing a rheumatologist or other specialist for their joints and none of the subjects reported joint pain at the time of testing. Surface electromyography (EMG) was used to record muscle activity.

The analysis of the data was realized in a semi-quantitative mann

The analysis of the data was realized in a semi-quantitative manner, the scores presented a variation from “−” for no labelling to “+, ++ and +++” to less, moderate and intense labellings, respectively. As described in previous studies from our lab,11 and 12

estrous cycle was monitored and OVX/O and OVX/RLX group presented diestrus smear, atrophied uterine horns and lower plasmatic concentration of estradiol. In contrast, the animals submitted to sham surgery presented the four regular stages of the selleck inhibitor estrous cycle, and the animals of group OVX/E2 presented enucleated cornified cells. For all experimental groups, positive immunolabelling for OPG and RANKL protein were visualized in cells of connective tissue, osteoblasts around the trabeculae bone and in osteocytes

aprisioned in the bone tissue formed during the alveolar healing process. TRAP protein was observed in osteoclasts present around the alveolar walls and close to the neoformed trabeculae bone. At 7 postoperative days, besides the great amount of haemosiderin, it was observed discrete RANKL immunolabelling in osteoblasts around trabeculae bone and osteocytes of the middle third (Fig. 1). Fibroblasts of the connective tissue presented moderate immunolabelling Thiazovivin datasheet of OPG protein (Fig. 2). OVX/O group presented the highest immunolabelling for OPG and RANKL protein than the other groups. TRAP immunolabelling were not visualized in the middle third, only a discrete labelling

in the borders of the dental Acyl CoA dehydrogenase socket with no significant difference between the groups (Fig. 3). At 14 postoperative days, it was observed RANKL immunolabelling (Fig. 1) similar to the previous period of all groups. Sham and OVX/RLX groups showed similar OPG immunolabelling (Fig. 2) compared to the previous analysed period, whilst OVX/O and OVX/E2 showed a decreasing of OPG immunolabelling. No background labelling with haemosiderin was observed which facilitates the visualization of the area. OVX/O group showed intense TRAP immunolabelling, moderate for OVX/E2 group and discrete for sham and OVX/RLX groups (Fig. 3). At 21 postoperative days, OVX/O group showed a decreasing OPG immunolabelling whilst it was increased for OVX/RLX group compared to the previous period (Fig. 2). Additionally, an increasing of RANKL immunolabelling was observed for all experimental groups (Fig. 1). These findings suggest an increasing in the cellular activity of bone remodelling process in order to form bone tissue in the presence of raloxifene. Considering TRAP immunolabelling, OVX/O group showed an intense expression, OVX/E2 group showed a moderate expression whilst sham and OVX/RLX showed a discrete expression, similar to previous analysed period (Fig. 3).

Continuous variables were compared using Wilcoxon rank sum test (

Continuous variables were compared using Wilcoxon rank sum test (since non-normally distributed). Characteristics of the tests were analysed in 2 x 2 tables using the ‘diagt’ routine.20 The McNemar test was used to compare sensitivities of the tests. Two hundred and twenty nine patients were examined. All patients met the WHO clinical case definition for acute dengue infection.21 One hundred and sixty two patients (71%) returned for the follow-up specimen collection visit and were considered further. Of the 162 patients

included in the current analysis, 72 patients (44%) were given a laboratory confirmed diagnosis of dengue infection by paired serology (Table 1). Four patients were found to have evidence of recent dengue infection. The demographic and clinical data of the patients are shown in Table 2. Patients with confirmed dengue infection had lower white cell counts (4.8 vs 7.2, P<0.0001) learn more and platelets (147 vs. 162, P=0.03) than patients with non-dengue infection. Twenty six patients (16%) were found to have non-dengue causes for their illness: four patients (2%) grew Salmonella typhi from blood cultures, by serology nine patients (6%) had murine typhus, seven (4%) had scrub typhus, and six (4%) had leptospirosis.

Only one patient had evidence of dual infection: acute secondary dengue and typhoid; this patient had positive NS-1 antigen and dengue rRT-PCR and also grew Salmonella typhi from a blood Selumetinib clinical trial culture. The serotype of about dengue virus was sought by performing nested RT-PCR on the acute specimens from the 72 serologically

confirmed cases. Seventy one of the cases (99%) were serotype 3, with the remaining case serotype 2. The four patients with serological evidence of recent dengue infection were negative in the nested RT-PCR. The performance characteristics of NS-1 antigen detection, rRT-PCR, and IgM antibody detection in the acute plasma specimen against our ‘gold standard’ of paired serology are shown in Table 3. NS-1 antigen or IgM antibody test alone had low sensitivity compared with the paired serology result, 54% and 17% respectively. rRT-PCR sensitivity was 89%, significantly higher than both NS-1 antigen and IgM antibody tests (P<0.00001). Specificities of NS-1 antigen detection, rRT-PCR, and IgM antibody detection were 100%, 96% and 88% respectively. The effect of fever duration at presentation on assay sensitivity is shown in Figure 1. NS-1 antigen sensitivity peaked in the early stages of fever (three days of fever at presentation). IgM antibody sensitivity rose later, peaking in patients presenting with five days of fever. The sensitivity of rRT-PCR remained high throughout. However, as a result of the relatively small number of specimens on each day, particularly for four and five days of fever, the 95% confidence intervals around the sensitivities are wide. The performance characteristics of combinations of the acute specimen tests are shown in Table 3 (combined by an ‘OR’ operator).

36 and 37 All 5 studies contributed individual patient data to th

36 and 37 All 5 studies contributed individual patient data to the Selleckchem Bcl-2 inhibitor GERD control group and 4 of the studies contributed individual patient data to the population-based control group. Study-specific definitions of the case and control groups are detailed in Table 1. In total, the 5 studies provided 1320 cases of Barrett’s esophagus, 1659 GERD controls, and 1434 population-based controls. For this analysis, and if a study provided such data, we excluded individuals who had ever smoked pipe tobacco or cigars (156 Barrett’s esophagus cases, 132 GERD controls, 153 population-based

controls) because comparing cigarette smokers with those who do not use other forms of tobacco provides a more accurate estimate of the effect of cigarette smoking. Ever smoking pipe tobacco or cigars was defined as meeting a study-specific low JAK inhibitor threshold exposure (a period of ≥6 months or ≥20 times during the life-course). Because of the relatively small number of non-white Barrett’s esophagus cases remaining

(17 black, 31 Hispanic, 39 other, and 18 missing), we restricted our analysis to white study participants. After exclusions, there remained 1059 Barrett’s esophagus cases, 1332 GERD controls, and 1143 population-based controls for analysis. Data acquisition and data pooling for each study were approved by the Institutional Review Board or Research Ethics Committee of the institute(s) sponsoring the study. The primary exposure variables were cigarette smoking status (ever vs never) and total cigarette smoking exposure (pack-years; 0, <15, 15–29, 30–44, ≥45). Additional exposure variables included duration of cigarette smoking (<30 years, ≥30 years), cigarette smoking intensity (<1, 1, and >1 packs/day), age of cigarette smoking initiation (<17, ≥17 years), and duration of cigarette smoking cessation (<20 years, ≥20 years). Cigarette smoking intensity and cigarette smoking duration in the University of North Carolina-Chapel Hill study were ascertained in categories and were recoded to the median

of the categories using the distributions of the other 4 studies combined. Ever Ergoloid cigarette smoking was defined as either low threshold exposures (≥100 cigarettes, ≥20 packs of cigarettes, 1 cigarette a day for ≥6 months) or by asking whether the patient had ever smoked. The following covariates were assessed for inclusion in regression models: age; sex; BMI (weight divided by square of height [kg/m2]); education; alcohol; fat, and trans-fat consumption; calories per day; meat, vegetable, and fruit servings per day; fiber consumption; heartburn, and regurgitation (population-based control models only); esophagitis; Helicobacter pylori seropositivity; hiatal hernia; and medication use (ie, nonsteroidal anti-inflammatory drugs, antacids, proton pump inhibitors, and H2-receptor antagonists).

This will lead to an inverted “U”-shape, which was observed along

This will lead to an inverted “U”-shape, which was observed along with extracellular hydrohalite shells as opposed to the linear correlation in case of intracellular hydrohalite formation. We recorded 24 confocal Raman images as the one shown in Fig. 1e distributed on four different samples containing L929 mouse fibroblast cells without Me2SO. All images except one contain hydrohalite found over the entire sample. The last image does not contain hydrohalite. We also investigated 6 samples with Me2SO, but only found a significant amount of hydrohalite in one, of which we recorded 6 Raman images. Each Raman image contained primarily one cell, but images with

up to three cells were

also recorded. All samples were subjected to identical freezing protocols. A typical transmission Akt inhibitor (TM) image and the corresponding Raman responses from cellular matter and hydrohalite are shown in Fig. 1b–d. These images contain one cell and an interdendritic channel. This can however not directly be concluded from the TM image alone. The Raman images reveal that the dendritic channel contains a high amount of hydrohalite and no cellular C646 concentration matter, whereas the hydrohalite phase overlaps the Raman response from the cellular matter. It can furthermore be concluded from the Raman images that the investigated cell contains a large aminophylline intracellular ice crystal, since most of the cellular matter is displaced towards the rim of the cell, and this displacement can only be attributed to intracellular ice crystals. These features cannot readily be seen from the TM image and clearly demonstrates how Raman imaging gives both more structural and chemical information compared to conventional imaging techniques. We found that the recorded Raman images of the samples without Me2SO can be roughly divided into three different classes, exemplified by the Raman images in Fig.

3a–c. Overlaps between the groups do however exist and some images are attributed to multiple classes. The first class, denoted Class A, contains images with very little or no overlap between the hydrohalite phase and cellular matter. This can readily be seen in the Raman images as in Fig. 3a. The hydrohalite in these images are thus clearly extracellular, although in close proximity to the cell. We found that 6 images out of the 24 contained extracellular hydrohalite. The two remaining classes, denoted Class B and Class C, contain Raman images with overlapping hydrohalite phase and cellular matter, i.e. data points where the focal volume contains both hydrohalite and cellular matter. Class B is defined to contain intracellular hydrohalite whereas the hydrohalite is located outside the cell for Class C. Two more examples of recorded Raman images are shown in Fig.