In order to get a clearer view on the precise function of the pro

In order to get a clearer view on the precise function of the processes underlying familiar and unfamiliar sequences ERK phosphorylation it seems better to separate motor preparation from motor execution. Therefore a modified version of the DSP-task was developed, inspired by the precuing paradigm of Rosenbaum (1980). In Rosenbaum’s paradigm precues (S1) provide specific information about the forthcoming movement. After a delay period an execution/withhold (go/nogo) signal (S2) is presented, which may provide missing information about the forthcoming movement in case of partial or non-informative precues or simply a go/nogo signal. Similar to the S1–S2 paradigm of Rosenbaum,

a go/nogo version of the DSP task was designed in which six key-specific stimuli were presented in sequence, which after a preparatory interval were followed by a go/nogo signal. In case of a go signal, participants were to react as fast and accurately as possible by pressing the six corresponding keys in the indicated order, and in case of a nogo signal responses should be withheld. This modified DSP task allows us to study the preparation phase of sequence learning in isolation from motor execution. To study movement preparation measures derived from the EEG appear especially useful (Dirnberger et al., 2000, Van der Lubbe et al., 2000 and Verleger et al., 2000). Event related potentials (ERPs) are indeed suitable to track the time course of functional processes underlying

movement preparation. In the present study, we employed the contingent negative variation (CNV), the lateralized readiness potential (LRP), and the contralateral delay activity (CDA) to study preparation MS-275 price of motoric sequences, since they give information about several PI-1840 different aspects of preparation. The CNV is a negative going wave with mostly a central maximum that unfolds in the interval between a warning stimulus and an execution signal (e.g. a go/nogo signal) (Jentzsch and Leuthold, 2002 and Verleger et al., 2000). The late CNV is typically maximal at the

Cz electrode and is thought to reflect preparatory motor activity (cf. Brunia, 2004 and Schröter and Leuthold, 2009). What exactly is represented in the CNV is unclear. Cui et al. (2000) suggest that the complexity of the prepared response is reflected in the CNV. In their study a simple and complex motor task were compared. During the simple movement task thumbs were opposing the index fingers three times in a row, by both hands. The complex movement task was the same, except that the second thumb oppositions involved the little fingers instead of the index. An increased late CNV for complex movements as compared with simple movements was obtained, which suggests that more preprogramming is taking place before complex movements compared with simple movements. In contrast with Cui et al., 2000 and Schröter and Leuthold, 2009 suggest that the amount of prepared responses is reflected in the CNV.

Regardless of the category of the quality characteristic, a great

Regardless of the category of the quality characteristic, a greater S/N ratio corresponded to better quality characteristics ZD1839 datasheet [18]. The method of calculating the S/N ratio depends at each run of the experiment on whether the quality characteristic is lower-the-better, higher-the-better, or nominal-the-better [30]. Accordingly, the three cases with respective equations are narrated below: (a) Upper-bound effectiveness (i.e., higher-the-better) equation(1) SN ratio=−10log(1n∑i=1n1yij2)where y

 ij = i  th replicate of j  th response, n=numberofreplicates=1,2,⋯,n;j=1,2,⋯,k.n=numberofreplicates=1,2,⋯,n;j=1,2,⋯,k. Eq. (1) is applied for problem where maximization

of the quality characteristic of interest is required. (b) Lower-bound effectiveness (i.e., lower-the-better) equation(2) SN ratio=−10log(1n∑i=1nyij2)Eq. (2) is applied for the problem where minimization of the quality characteristic is required. (c) Moderate effectiveness (i.e., nominal-the-best) equation(3) SNratio=10log(y¯2s2)where, y¯=y1+y2+y3⋯+ynnand s2=Σ(yi−y¯)2n−1 A nominal-the-best type of problem is one where minimization of the mean squared error around a specific selleck chemicals target value is desired. Adjusting the mean on target by any means renders the problem to a constrained optimization problem. This sub-section illustrates step-by-step the theory and methodology of GRA. Step 1: Calculated the S/N ratios for the corresponding responses using one of the formulae (Eqs. (1), (2) and (3)) depending upon the type of quality characteristic. Step 2: Normalized the Yij as Zij (0 ≤ Zij ≤ 1) by the following formula to avoid the effect of using different units and to reduce variability. The normalization is a transformation performed on a single input to distribute the data evenly and scale it into acceptable range for further analysis. Haq et al. [12] recommended that the S/N ratio should be used to normalize the

data in GRA. For further analysis, normalization is applied on each response to distribute the data evenly and in acceptable range [7]. equation(4) Zij=Yij−min(Yij,i=1,2,⋯,n)max(Yij,i=1,2,⋯,n)−min(Yij,i=1,2,⋯,n)Eq. (4) was CYTH4 used for the S/N ratio with higher-the-better case. equation(5) Zij=max(Yij,i=1,2,⋯,n)−Yijmax(Yij,i=1,2,⋯,n)−min(Yij,i=1,2,⋯,n)Eq. (5) was used for the S/N ratio with lower-the-better case. equation(6) Zij=|Yij−Target|−min(|Yij−Target|,i=1,2,⋯,n)max(|Yij−Target|,i=1,2,⋯,n)−min(|Yij−Target|,i=1,2,⋯,n)Eq. (6) is applicable for the S/N ratio with nominal-the-better case. Step 3: Determined quality loss functions by using the eq. Δ = (quality loss) = |yo−yij||yo−yij|. Step 4: Computed the grey relational coefficient (GC) for the normalized S/N ratio values.

Moreover it is relevant to make the diagnosis for the clinician,

Moreover it is relevant to make the diagnosis for the clinician, since this lesion is highly prone to induce thrombus formation on its surface, with

the possibility of embolic events. Early CEA is recommended and it is again relevant Rapamycin supplier for the surgeon to suspect this diagnosis since, if the lesion is not completely removed, it can grow back again, with the risk of further embolic events. “
“Since the work of Call and Fleming in 1988 [1] a variety of similar syndromes with reversible cerebral vasoconstriction were published. Today these syndromes are unified in the term reversible cerebral vasoconstriction syndrome [2]. According to literature the reversible cerebral vasoconstriction syndrome is characterized by the following facts. The mean age of onset is 42 years. Women are affected 2–3 times more

often than men [5]. The syndrome is associated with pregnancy and puerperium, drugs such as cocaine, cannabis, LSD, ergotamine or selective serotonin reuptake inhibitors, different types of headache such as migraine, primary thunderclap headache, primary headache associated ALK inhibitor cancer with sexual activity and other conditions such as porphyria, pheochromocytoma, craniocerebral injury [3] and [4]. According to the work of Ducros et al. the main clinical manifestation in 94% of 67 patients were thunderclap headache recurring over a mean period of one week. Other symptoms were nausea, vomiting, confusion and blurred vision. 3% of the patients in Chlormezanone this review showed seizures [5]. Several vascular complications are reported. According to the work of Ducros 22% of the patients developed subarachnoidal hemorrhage, 6% intracerebral hemorrhage, 14% showed transient ischemic symptoms and 4% developed cerebral infarction in the course of disease [5]. Neuroimaging shows diffuse, multiple stenosis and dilatation of the cerebral vessels (string and beads) which resolve spontaneously in 1–3 months. There are no common transcranial color coded ultrasound criteria for diagnosis.

Therefore common criteria for intracerebral stenosis or vasospamus are used. Ultrasound is shown to be safe in diagnosing and in controlling the course of disease [6]. There is no standard treatment. Due to literature mainly the calcium antagonist nimodipine in systemic application or in some case reports in local application is used. The disease is self-limiting and has a low incidence of recurrence. But for prolonged vasoconstriction a higher risk of posterior leukencephalopathy and strokes is reported [6]. We report the case of a 32 year old primipara. The patient was admitted to an academical hospital with maximum medical care. The cause of admission was preeclampsia. For gynecological reasons a Ceasarean section (C-section) was necessary.

The acceptance test was carried out with 60 consumers (aged 21–50

The acceptance test was carried out with 60 consumers (aged 21–50 years), preselected according to interest and habits of cheese consumption. Consumer evaluation was performed check details according to a hedonic scale ranging from 1 (dislike very much) and 9 (like very much) for aspect, odor, texture, taste and overall appreciation. The testing sessions (trained panel and consumer testing) were conducted in individuals booths under conditions in accordance with ISO 8589 (facilities) and ISO11037 (lighting). Each assessor was served of 20 g of each

cheese sample placed on small white plates coded with three-digit random numbers served immediately after being taken out of refrigerated storage. Assessors were asked to use low-salt crackers and water to clean their palates between the assessed samples. Data acquisition was achieved by informatics system Fizz. All analyses were INK 128 cost carried out in triplicate. The means of the results were evaluated using analysis of variance (ANOVA), and Tukey’s test was used to compare significant differences (P < 0.05) between the physicochemical,

fatty acid profile, textural and sensory evaluations. The statistics model of sensory analysis data contained only a fixed effect of treatment. SPSS (v. 17, Chicago IL, USA) was used for the statistical analyses. The physicochemical characteristics of Coalho cheese made from cow’s milk, goat’s milk, and their mixture are shown in Table 1 and Fig. 1. In general, the moisture and salt contents were the highest (P < 0.05) in CCM. No significant difference (P > 0.05) was observed in protein content and in pH values regardless the type of cheese.

The fat content of CCGM and CGM were higher (P < 0.05) than Thymidylate synthase those of CCM for all the evaluated storage times. So, it is important to highlight that the reduction of goat milk to 50% did not affected any of the physicochemical parameters using Coalho cheese technology. Sheehan et al. (2009) studied the partial or total substitution of bovine for caprine milk during cheese production and showed that increased ratios of bovine:caprine milks resulted in cheeses with increased moisture, fat and fat-in-dry matter (FDM) contents with no significant effect on cheese protein, moisture-in nonfat-substance (MNFS) or salt contents. The significant effect observed for moisture and fat by these authors, but not for our CCGM cheeses, may be related with different technology used. The moisture, fat, salt and pH value found in CCM and CGM were similar to those reported by Pappa, Kandarakis, Anifantakis, and Zerfiridis (2006) who assessed the influence of type of milk (goat’s, ewe’s and cow’s milk) and microbial culture on the quality of Teleme cheese.

No entanto, os grupos não eram homogéneos para esta variável o qu

No entanto, os grupos não eram homogéneos para esta variável o que pode ser um viés de interpretação destes resultados. Admitimos portanto que o ensino personalizado poderá melhorar a preparação intestinal também nestas situações. Nos doentes diabéticos também não se verificaram diferenças significativas entre os grupos em relação à qualidade da preparação intestinal. Aqui, a intervenção personalizada ocorreu no tipo de alimentos adaptados ao gosto do doente, mas Gemcitabine solubility dmso não num aumento da duração da dieta. Poderá ser tentada uma estratégia deste tipo, uma vez que muitas vezes esta patologia cursa

com aumento do tempo de trânsito intestinal e obstipação. Alcançou-se melhoria dos resultados nos doentes com escolaridade superior ao Ensino básico. this website Este facto poderá estar relacionado com uma maior capacidade intelectual do doente, permitindo uma melhor compreensão e adesão às medidas propostas sendo, no entanto, discutível esta interpretação, dado que doentes com escolaridade inferior podem ter compreensão e adesão igualmente semelhantes ou o apoio de outros familiares no cumprimento das orientações dadas. O nosso estudo apresenta limitações que devem ser referidas. Uma é o tamanho da amostra, dado termos concluído serem necessários

pelo menos 199 doentes em cada grupo de estudo para a intervenção ter o efeito pretendido e estarem incluídos apenas 67 doentes no grupo «controlo» e 58 no grupo «intervenção». De facto, estes resultados são ainda preliminares e o ensaio prossegue no nosso Serviço, tendo-se decidido fazer uma avaliação a esta altura por se ter incluído cerca de um terço dos doentes. Por outro lado, seria importante valorizar o impacto que a melhoria da qualidade da preparação intestinal tem na

eficácia da colonoscopia, em função do tempo de cada exame, da percentagem de deteção de lesões e do custo, e essa análise não foi efetuada. De facto, apesar de o ensino personalizado poder melhorar a qualidade da preparação intestinal, esta estratégia obriga a encargos acrescidos, uma vez que exige a intervenção de um profissional de saúde por um tempo prolongado, e esse preço real não foi determinado. Para se tirarem conclusões fidedignas teria de ser confrontado com a eventual diminuição de custos, originada pela melhoria da preparação, ao aumentar a deteção de lesões permitindo um aumento dos intervalos buy Gemcitabine de vigilância, uma diminuição do número de exames de controlo e um alívio na sobrecarga das listas de espera dos serviços. Em conclusão, estes resultados sugerem que o ensino personalizado poderá ter importância na melhoria da qualidade da preparação intestinal na colonoscopia. Por ser um resultado preliminar, a generalização dos resultados ainda é incerta, pelo que, de momento, não podemos recomendar esta estratégia em todos os doentes. No entanto, em grupos selecionados, esta medida poderá ter um impacto positivo, justificando-se assim a sua utilização na prática clínica.

g 51 and 52]) It is interesting then to note that navigation is

g. 51 and 52]). It is interesting then to note that navigation is not dissimilar to the inverse of path integration: the former requires the calculation of the vector between two allocentric locations, while the latter uses recent motion,

expressed as a vector, to update an allocentric representation of self-location. As such it seems possible that the neural architecture that supports path integration might also play a role in navigation. Indeed, several authors have recently proposed models of navigation in which grid cells are seen as the central component http://www.selleckchem.com/products/pirfenidone.html of a network able to determine the allocentric vector between an animal’s current location and a remembered goal 53, 54 and 55]. However, the mechanisms employed by the models differ markedly, ranging this website from an iterative search for the appropriate vector [53] to a complex representation of all possible vectors projected into to the cyclic grid space [54]. As such, at the neural level, it is still too early to predict how the activity of individual grid cells might be modulated during navigation. However, at the population level accessible to fMRI, it seems plausible that metabolic activity in the entorhinal cortex should correlate with allocentric spatial parameters. Indeed it is already known that the coherence of the directional

signal associated with grid cells correlates with navigational performance [56]. Furthermore, in light of the limitations imposed on place cell models of navigation by the irregular distribution of place fields, it seems Carnitine palmitoyltransferase II more likely that activity in the hippocampus will reflect route based variables. A number of recent fMRI studies have examined whether brain activity is correlated with the distance between landmarks or to goals during navigation. During navigation a number of spatial parameters represent the navigator’s relationship to the goal (Figure 2a) and these parameters change over the different key events

and epochs that characterise navigation (Figure 2b). Humans have been shown to be reasonably good at estimating parameters such as Euclidean distance, path distance, and direction to distant locations, at least in large complex buildings [57]. Two studies have reported increased activity in the mid to anterior hippocampus at the start of navigation when route planning was required 8 and 58]. Such activity may relate to the initial demands of planning the route to the goal, however it was not clear whether this activity was related to the distance to the goal. The first fMRI study to examine spatial goal coding found that activity in the entorhinal cortex of London taxi drivers was significantly positively correlated with the Euclidean distance to the goal during the navigation of a virtual simulation of London, UK [9•] (Figure 3a). This result is consistent with the entorhinal cortex coding an allocentric vector to the goal 53, 54, 55 and 59]. Several recent studies have adopted a similar approach (Figure 3b–d).

As noted, another limitation is that the 12-month study period wa

As noted, another limitation is that the 12-month study period was too short to adequately capture improvements in pediatric-specific parameters such as puberty (as evaluated

by Tanner stage) and bone mineral density analysis. However, these parameters will continue to be followed in extension study PB-06-006 (NCT01411228) that will capture an additional 2 years Transmembrane Transporters activator of data for a total of 3 years of taliglucerase alfa treatment. In summary, this report demonstrates that taliglucerase alfa improves the hematologic and visceral manifestations of Gaucher disease in children. It broadens the findings to date of the safety and efficacy of taliglucerase alfa in patients with GD, pediatric and adult patients alike, and as such expands the potential treatment options for management of this genetic metabolic disorder. AZ designed the study, performed research, analyzed data, and wrote the paper; DEG-R performed research and wrote the paper; AA performed research and wrote the paper; DE assisted

this website with the research and wrote the paper; AP designed the study, analyzed and verified data, and wrote the paper; EB-A designed the study, analyzed and verified data, and wrote the paper; and RC designed the study, analyzed and verified data, and wrote the paper. None of the authors received compensation for their contributions to this manuscript. AZ receives consultancy fees from and Diflunisal has stock options in Protalix BioTherapeutics and is a member of their Scientific Advisory Board. In addition, AZ receives support from Genzyme for participation in the International Collaborative Gaucher Group Registry, and receives honoraria from Shire HGT, Actelion, and Pfizer; DEG-R and AA are study investigators; DE has received honoraria from and had travel/accommodation expenses covered/reimbursed by Shire HGT and Pfizer. In addition, the Gaucher Clinic, for which DE is the site coordinator, has had clinical trial expenses reimbursed; AP, EB-A, and RC are employees of Protalix BioTherapeutics. The authors would like to acknowledge fellow

investigator and pediatrician Dr. Rene Heitner from Johannesburg, South Africa, who passed away in January 2012. The authors would also like to acknowledge Dr. Peter Cooper of Johannesburg, South Africa, who is treating Dr. Rene Heitner’s patients in study PB-06-006, the taliglucerase alfa pediatric extension trial. This study was sponsored by Protalix BioTherapeutics. Editorial and medical writing support was provided by Elizabeth Daro-Kaftan, PhD, of Peloton Advantage, LLC, and was funded by Pfizer. Pfizer and Protalix entered into an agreement in November 2009 to develop and commercialize taliglucerase alfa. “
“Acute Myeloid Leukemia (AML) is primarily a hematological malignancy of the elderly with a median age of onset at 60 years and a poor prognosis with a five year survival rate of only 12% [1].

Ultimately, we hope that this paper will stimulate new perspectiv

Ultimately, we hope that this paper will stimulate new perspectives in order to access and assess (self) awareness also in clinical populations such as DOC patients. A sample consisting of 14 subjects (9 females, 5 males) with age ranging from 21 to 53 (M=25.79; SD=8.17) was recorded. All volunteers were right-handed German native speakers without any recorded history of neurological disease. Participants gave written

informed consent approved by the local ethics committee and received monetary compensation for their participation. The experiment expands the SON task as introduced by Schnakers et al. (2008) and subsequently adapted in Fellinger et al. (2011). Stimuli were either spoken by a familiar (FV; subject’s close friend or family member) or unfamiliar voice (UFV; spoken by a text-to-speech algorithm, Alectinib CereProc®, CareProc Ltd: Olaparib cost “Alex”, “Gudrun”). Stimuli included the subject’s own name and five commonly used Austrian names (according to statistics Austria) matched for number of syllables and the gender of the participant. Stimuli were presented via headphones at a sound pressure level of 80 db. The task consisted of two experimental conditions: an active condition to investigate the ability to consciously follow commands and a passive listening condition

with the passive condition always preceding the active condition. Each condition consisted of 3 blocks; with each block including 13 presentations of each name (i.e., 39 presentations for each single name). In the passive condition 6 stimuli were presented with 234 repetitions in total (about 12 min), in particular, SON uttered by a familiar or unfamiliar voice and two different unfamiliar names either spoken by a familiar or an unfamiliar voice. In the active condition

only 3 different stimuli were presented (117 repetition) for about 6 min, all of them unfamiliar to participants and all uttered by a familiar voice (cf. Fig. 1). During the passive condition participants were simply asked to listen to all the names presented, while in the active condition they were asked to focus and silently count the appearance of the target name. In order to be sure that participants attended the presented stimuli experimenters Rebamipide controlled at the end of the experiment whether the number of targets counted by participants matched the total number of stimuli presented and controlled online for arousal fluctuations. The inter stimulus interval [ISI] lasted 2000 ms and for stimulus presentation and synchronization, the Software Presentation®, (Version 0.71; Presentation Software, Neurobehavioralsystems Inc., CA) was used. EEG was recorded with 32 Ag/AgCl sintered electrodes and head circumference matched Easycaps (EASYCAP GmbH; Herrsching Germany) placed according to the international 10–20 system.

Importantly, the ER assessment is almost instantaneous and highly

Importantly, the ER assessment is almost instantaneous and highly suited for static Franz-type diffusion cells. The magnitude of change per 5, 10 or greater number tape strips differed among the skin integrity indices measured. A further analysis of the data where the changes were compared with those observed for TEWL in clinical situations revealed that removal of 10 tape strips provided a loss of barrier function approximately equivalent to a 3–4-fold increase in TEWL in

vivo, while also providing a discernible decrease in ER. This 3–4-fold increase in TEWL approximates Antidiabetic Compound Library to the altered barrier function observed clinically in atopic dermatitis, psoriasis, and diaper dermatitis as described previously ( Goon et al., 2004, Kim et al., 2006 and Stamatas et al., 2011). The experimental work presented here has shown

that the removal of 10 tape strips is the most relevant procedure for this in vitro skin model in order to make realistic predictions of skin penetration in compromised skin. We recognise that all three measurements (TEWL, TWF and ER) can be utilised to determine BI 2536 manufacturer whether skin barrier function has been compromised to a standardised level. Indeed, it may be appropriate to combine different measures depending on the circumstances being investigated. For example, if a skin application was designed to prevent water loss then the TEWL approach may be better to assess performance and this method, of course, can be run in parallel in clinical investigations. One area where we think this in vitro methodology would be useful is for the safety assessment of new and existing consumer and pharmaceutical

products. There Oxymatrine is little information in the area of dermal penetration of topical drug and cosmetic formulations under conditions where the stratum corneum is damaged, diseased or even absent, such as following sunburn. The risk assessment process may incorrectly assume that the systemic exposure to a drug, for example, is perhaps ten times higher when the skin barrier is impaired. However, this may be a gross over-estimate for most compounds. It is obviously an area of safety assessment where the ethical considerations would not justify investigation of this effect in animals or humans. Therefore, the scientifically-based approach we have presented here using ex vivo skin and ER is a step forward in this area of dermatokinetics to aid the risk assessment process where exposure is to a compromised skin barrier. Clearly, further investigation is required to establish whether there is a clear link to the physicochemical properties of the compound in question or the vehicle and the formulation in which it is applied. This may lead eventually to mathematical prediction models similar to those used for dermal absorption through normal skin. The authors declare that there are no conflicts of interest. Transparency Document.

However, the ratio of annexin-V positive to negative MV was not s

However, the ratio of annexin-V positive to negative MV was not sensitive to anticoagulant (r2 = 0.08). MV recovery was the same from blood collected in Vacutainer or non-Vacutainer tubes containing the same concentration of calcium chelating and protease inhibitor click here anticoagulants (not shown). Does calcium chelation suppress MV recovery or do protease inhibitors stimulate shedding? The results with

endothelial MV suggest suppression. We had observed that there was a window of as long as 10 min between phlebotomy and mixing of the blood with anticoagulant during which the MV count was stable. Accordingly, blood (1 mL aliquots) without anticoagulant was centrifuged immediately for 2 min Thiazovivin supplier at 8000 × g or for 10 min at 3000 × g, and then anticoagulants were added to these platelet poor plasmas (PPP). Addition of any anticoagulant to PPP thus prepared from non-anticoagulated blood yielded the same number of annexin-V positive MV as blood collected in H&S anticoagulant ( Fig. 3). The basis for the loss of MV with removal of calcium was addressed by shifting the point of addition of anticoagulants. Adding either calcium chelating or protease inhibitor anticoagulants to isolated MV did not alter MV counts (data not shown). When

calcium chelators were added to the platelet rich plasma (PRP) prepared from the first 800 × g spin of blood collected in H&S or heparin ( Fig. 4, top), platelet MV counts decreased to an extent similar to that seen in whole blood with citrate or EDTA anticoagulants ( Fig. 4, bottom). In contrast, addition of H&S or heparin to PRP prepared from blood collected in calcium chelating anticoagulants did not further affect numbers of MV ( Fig. 4, bottom). Whole blood collected in either citrate or H&S was Methocarbamol distributed into 1.5 mL tubes and maintained at either room temperature (ca. 22 °C) or 33 °C for up to 3 h, during which MV counts were obtained at intervals. For whole blood collected in citrate, counts

of annexin-V positive and platelet MV decreased within 15 min and were significantly lower after one hour at either temperature (data not shown). In contrast, counts of annexin-V and platelet MV did not change significantly within the first hour at either temperature in blood collected in H&S but increased significantly thereafter (Fig. 5). The increase in counts of stained MV was greater at room temperature than at 33 °C. However, the percentage of platelets expressing surface P-selectin, activated glycoprotein αIIbβ3, phosphatidylserine remained < 5% in all samples. Counts of endothelial MV did not change during the three hours at either temperature. Centrifugation of PFP at 20,000 × g recovered on average 80% of the MV measured by direct staining of PFP (r2 = 0.8). More than 90% of platelet MV were recovered after a wash with Hanks’/HEPES of MV pelleted by the 20,000 × g centrifugation (n = 66).