In what follows the reader is provided with a brief overview of t

In what follows the reader is provided with a brief overview of the clinical evidence of pain physiology education in patients with chronic musculoskeletal pain. The largest part of the paper is dedicated to practice guidelines on how to apply pain physiology education in patients with chronic musculoskeletal pain. Several groups have studied the clinical effects of pain physiology education in various chronic musculoskeletal pain populations such as chronic low back pain (Moseley, 2002, Moseley, 2003b, Moseley, 2004, Moseley,

2005, Moseley, 2004 and Ryan et al., 2010), fibromyalgia (Ittersum et al., Submitted for publication, Ittersum van et al., in press and Van Oosterwijck et al., submitted for publication, chronic whiplash associated disorders (Van Oosterwijck et al., 2011) and chronic fatigue syndrome with chronic widespread PLX3397 concentration pain (Meeus et al., 2010a). In find more patients with chronic low back pain, pain physiology education alters pain perceptions and, in conjunction with physiotherapy, it improves functional and symptomatic outcomes (Moseley, 2002, Moseley, 2003b, Moseley et al., 2004 and Moseley, 2005). A recent randomized controlled trial indicates that, in the short term, pain physiology education alone is more effective

for pain relief and improving pain self-efficacy than a combination of pain physiology education and group exercise classes for patients with chronic low back pain (Ryan et al., 2010). Altered pain perceptions are directly associated with altered movement performance in those with chronic low back pain, even if there is no opportunity for the patients to be physically active during the treatment (Moseley, 2002 and Brosschot, 2002). This implies that motor performance may be directly limited by pain perceptions. Indeed, a case series study of patients with chronic whiplash associated disorders showed improvements

in illness perceptions, pain thresholds and movement performance (Van Oosterwijck et al., 2011). In patients with chronic fatigue syndrome, pain physiology education alters pain perceptions such as catastrophizing, and pain behaviour (Meeus et al., ID-8 2010a). In another randomized controlled clinical trial, we showed that simply providing the detailed information booklet explaining pain physiology and central sensitization, did not change illness perceptions or health status in patients with fibromyalgia (Ittersum et al. submitted for publication). However, when the same written education about pain physiology was combined with two educational sessions (one face-to-face session and one by telephone) of individually-tailored pain physiology education, vitality, physical functioning, mental and general health of patients with fibromyalgia improved (Van Oosterwijck et al. submitted for publication).

Extended exposure (14 weeks) to the lowest dose

(0 02 mg 

Extended exposure (14 weeks) to the lowest dose

(0.02 mg AP kg−1) gave similar results (Meier et al., 2011). These exposure levels are difficult to compare with real-life exposure to PW plumes, especially since many endocrine disruptors seem not to produce linear dose–response curves (Vandenberg et al., 2012), but the authors themselves consider the exposure level higher than what is realistic, possibly demonstrating a worst-case disturbance of reproductive fitness in the cod. Also, Sundt and Bjorkblom (2011) recorded impaired oocyte development and reduced estrogen levels in pre-spawning female Atlantic cod, as well as altered testicular development, an increase in the amount of spermatogonia and primary spermatocytes, and a reduction in the amount of mature sperm in males following exposure to realistic concentrations of PW (0.066–0.2%) for twelve weeks. Therefore, one cannot exclude that APs in PW effluents under certain circumstances could cause reproductive selleck chemicals llc disturbance in sensitive stages (e.g. pre-spawning) of wild fish that stay close to offshore platforms for long periods of time. However, it seems unlikely that this could JAK2 inhibitor drug affect a significant fraction

of Atlantic cod populations. Estrogens are involved in many biological processes, including control of gonad maturation in male and female fish. The enzyme cytochrome P450 aromatase converts androgens, like testosterone or androstenedione to estrogen (E2) and estrone. Sinomenine Teleost fish have two aromatase genes; one that is mainly expressed in the gonads (aromatase A or cyp19a1a), and one that is mainly expressed in the brain (aromatase B or cyp19a1b) ( Diotel et al., 2010). Meier et al. (2011) did not find any regulation of cyp19a1a in the ovary

(mRNA expression or enzyme activity), or of aromatase activity in the brain of female cod exposed to AP or PW. The specific activity of aromatase in the ovary was therefore not affected by the AP-exposure. Tollefsen et al. (2007) and Thomas et al. (2009) used recombinant yeast estrogen and androgen screens to determine the in vitro estrogen receptor (ER) agonist and androgen receptor (AR) antagonist potencies of solid phase extracts (SPE) of PW collected from 20 Norwegian installations. They found estrogenic activities at levels equivalent to <0.1–4 ng L−1 E2 (dependent on PW source), similar to those previously reported for the UK continental shelf (UKCS) ( Thomas et al., 2004). No activity was detected after exposure to filtered oil droplets from PW suggesting that ER activity was primarily associated with the dissolved phase. Thomas et al. (2009) identified short-chain petrogenic APs to be responsible for around 35% of estrogen receptor (ER) agonist activity measured in vitro. Androgen receptor (AR) antagonists were detected both in the dissolved and oil associated phase. They also reported that naphthenic acids, which occur in significantly higher concentrations than C4–C7 APs in PW, were weak ER agonists.

, 1997) could thereby be exacerbated, decreasing reproductive out

, 1997) could thereby be exacerbated, decreasing reproductive output in affected males. Conversely, reduced sperm swimming under acidified conditions could increase sperm longevity due to lowered consumption of limited endogenous energy provisioning ( Mita and Nakamura, 1998). Greater sperm longevity may increase chances of successful fertilization if sperm–egg-encounter rates remain sufficient over SGI-1776 in vivo prolonged periods of time ( Levitan, 2000 and Marshall, 2002). Impacts of CO2-driven ocean acidification on

sperm swimming behavior of G. caespitosa may interact with other acidification impacts on fertilization variables such as male–female compatibility, egg competition or polyspermy block efficiency ( Evans and Marshall, 2005, Evans and Marshall, 2005 and Marshall and Bolton, 2007). FK866 datasheet Negative impacts of CO2-induced ocean acidification have also been

reported for later life-history stages of serpulid tubeworms, such as weaker calcareous tubes ( Chan et al., 2012 and Smith et al., 2013). Resultant cumulative effects on reproductive success and survivorship are likely to exacerbate the rate or intensity of selection pressure of climate change. Patterns of sperm swimming responses of G. caespitosa to CO2-induced acidification observed here were similar to those of Arenicola marina sperm in lowered seawater pH ( Pacey et al., 1994). Sperm activation in A. marina was delayed and sperm speed was reduced in HCl-acidified seawater (pH < 7.6). Interestingly, our findings are very different to those from NADPH-cytochrome-c2 reductase a study on the related serpulid species Pomatoceros lamarckii ( Lewis et al., 2012). Sperm speeds of P. lamarckii were robust to CO2-induced pH reductions, percent motility was significantly reduced, but responses were non-linear. These findings may be explained by differences in experiment design and sample

size (5 pooled assays ( Lewis et al., 2012) vs 23 single individuals in this study). As outlined earlier, conducting adequately replicated studies will help to clarify whether these differences are caused by high inter-individual variability or differences in average responses between species. In conclusion, the substantial inter-individual variation in sperm responses observed here may ameliorate effects of future climate change, if the traits that drive phenotype robustness are heritable. Sperm from some G. caespitosa will be better equipped to cope with acidification than others, creating ‘winners’ and ‘losers’ in a future acidified ocean ( Schlegel et al., (2012). This observed resilience to near-future conditions could increase the potential for adaptation to far-future conditions, if gathering of advantageous alleles can occur quickly enough. Likewise, rapid selection against phenotypes susceptible to acidification may quickly reduce genetic diversity and lead to severe flow-on consequences for fitness and competitive ability downstream. Very few studies to date have investigated climate change impacts on polychaete species ( Chan et al.

While literature suggests that malnutrition and falls in frail el

While literature suggests that malnutrition and falls in frail elderly are related (Vellas et al., 1990 and Vellas et al., 1992), and a relation

via loss of muscle mass seems realistic, there are only a few empirical studies that investigated the relationship between fall incidents and nutritional status in this population. Daniels (2002) found that in residential care settings a substantial number of elderly susceptible to falling is also at risk of poor nutritional health. The primary aim of this study is to explore the relation between malnutrition and fallers in Dutch LTC residents. Residential LTC institutions in the Netherlands provide temporary or permanent multidisciplinary treatment, guidance, support and nursing care for elderly patients with long-term, complex health problems, expressed primarily in functional disorders and handicaps. Secondary, we will investigate Stem Cell Compound Library cell assay the role of activity within this relationship. Thirdly, we will investigate whether the relation between nutritional status and fallers is affected by Anti-diabetic Compound Library nutritional intervention. This study is a secondary data analysis of the annual independent National Prevalence Measurement of Care Problems of Maastricht University (LPZ called; in Dutch healthcare. Yearly, more than 400 health care organizations (hospitals, nursing homes, homes for the elderly, and home care organizations)

participate voluntarily in the LPZ measurement. It is a cross-sectional, multi-center point prevalence and incidence measurement. Patients are investigated regarding the prevalence or incidence, prevention, and treatment of several health care problems,

e.g. pressure ulcers, incontinence, restraints, intertrigo, falls and malnutrition. For this study we analyzed the Dutch malnutrition and falls data of 2008 (Halfens et al., 2008). In 81 LTC settings in the Netherlands, 6828 residents participated in the LPZ measurement regarding malnutrition and falls. The following exclusion criteria were applied: residents younger than 65 years, residents without complete data regarding fall history and/or nutritional status. Permission to conduct the study was obtained from the Medical Ethics Committee at Maastricht University Medical Forskolin cell line Center (MUMC). Prior to the data collection, oral or written informed consent by residents, relatives or legal guardians in case of psycho geriatric residents, preceded participation. The LPZ uses a standardized questionnaire to register amongst others data of measured weight, height, number of diseases, nutritional intake, undesired weight loss, nutritional interventions, fall history, Braden scale (Ayello & Braden, 2002), and Care Dependency Scale (CDS) (Dijkstra, Tiesinga, Platinga, Veltman, & Dassen, 2005). The Braden activity-item was used to score the amount of physical activity of the participants with the following categories: (1) bedfast, (2) chairfast, (3) walks occasionally, and (4) walks frequently.

, 2012 and Scott et al , 2010) The commercial aims of in vitro t

, 2012 and Scott et al., 2010). The commercial aims of in vitro testing are to be faster and cheaper, although currently, the costs are roughly on par Cytoskeletal Signaling inhibitor with Draize testing. It is preferable that the testing procedures can be performed without the need for specialist training or expensive equipment ( Dholakiya and Barile,

2013). From a corporate standpoint in vitro tests require the same level of investment as they are currently making using in vivo tests, so they either don’t care, or fail to see the benefits in switching. A large factor that affects the decision making of corporate companies is that they are selling to a local market, not just countries within the EU. For developing or newly industrialized countries, Brazil, Russia, India, China and South Africa, the underlying challenge is getting them to understand the roles of in vitro tests, which is a continuing educational challenge. In order to overcome these issues, a re-evaluation of currently used find more in vitro tests may be required ( Nóbrega et al., 2012). In vitro assays and models provide useful data that complement in vivo studies allowing for significant reductions in the numbers of animals used. In order realize this, it must be ensured that clear endpoints correlate

between in vivo and in vitro tests ( Maurer et al., 2002). In general, in vitro tests are validated against the Draize test ( Lenoir et al., 2011), with few actually investigating their predictability compared to humans. Despite the lack of formal validation, in vitro tests still are commonly used by industry. For example, industrial toxicologists often use in vitro protocols for prioritizing products and ingredients for further development ( Curren and Harbell, 2002).

However, use of the Draize test is still permitted worldwide, with the exception of the cosmetics section within Europe. Although in vitro alternatives tests are available, whether they are actually being used in practice is questionable. Every country has its own regulations and data requirements. The EU may be consolidated, but everywhere else is not and regulations have to be negotiated one by one – this is a very slow process, with STK38 no one country worse than the other. Regulations are aimed at protecting humans, and regulators focus on this, the culture of animal welfare is different in every country. In silico models are computer generated models that can play a useful role in predicting the ocular toxicity of a substance. In silico models utilize repositories of existing in vitro and in vivo toxicology data to predict the toxicity of samples. Quantitative structure–activity relationships (QSAR) are used to quantify the relationship between a sample’s chemical structure and the biological effects that result from the same chemical ( Simon-Hettich et al., 2006).

In prokaryotes, factors that package DNA, such as HU proteins, ma

In prokaryotes, factors that package DNA, such as HU proteins, may control supercoiling by binding to DNA and trapping the free energy of supercoiling as writhe and subsequently releasing it through controlled dissociation [ 3 and 4]. Similarly in eukaryotes the regulated release of terminal DNA from a nucleosome, mediated by the acetylation of core histone tails, could release constrained writhe for conversion into negative supercoiling. Although in

vitro studies support this concept [ 5] its operation in vivo is elusive [ 6]. In prokaryotes and eukaryotes all activities buy I-BET-762 that require DNA to be unwound (and rewound) are potent generators of supercoiling. The classic example is the ‘twin supercoiled domain’ model where elongating RNA polymerase, in unwinding the DNA, generates positive supercoiling ahead and, in rewinding the DNA, generates negative supercoiling in its wake [7 and 8] (Figure 1). The levels of supercoiling produced in this process are prodigious, amounting to a positive and a negative

supercoil for every 10 bp transcribed. Apitolisib molecular weight Consequently the role of topoisomerases in releasing torsional stress is crucial if the template is to be maintained in a transcriptionally competent state. Genes that are negatively supercoiled are generally more efficiently transcribed [9 and 10] but topoisomerase inhibition studies [11, 12•, 13 and 14] indicate that the accumulation of excessive positive or negative supercoiling will repress transcription. Therefore, there must be a regulated balance in the localised levels of supercoiling through the concerted actions of polymerases [15]

and topoisomerases [16 and 17]. When an activity supercoils Clomifene DNA the torque generated is transmitted along the molecule. If the ends of the molecule are not fixed (or at least hindered), the supercoiling will dissipate via the unhindered rotation of the helix. Therefore for supercoiling to have a structural or functional influence on DNA or chromatin it must operate within a constrained environment where the energy is at least transiently trapped or restricted. For this reason it is anticipated that genomes need to be organised into supercoiling domains with barriers that prevent the spread of topological stress. In prokaryotes the Escherichia coli genome has a hierarchical organisation based on large structural macrodomains [ 3] with the Ter domain being subdivided into smaller, 35 kb domains via MatS/MatP interactions [ 18]. This organisation establishes a dynamic structural architecture enabling packaging without interfering with transcription or replication. The genome is also separately organised into about 500 independent ∼10 kb supercoiling domains with demarcating barriers stochastically distributed and dynamically maintained [ 19 and 20].

Monocyte preparations were routinely stained with anti-CD14 antib

Monocyte preparations were routinely stained with anti-CD14 antibody (Becton Dickinson, Oxford, UK) followed by flow cytometric analysis to verify purity. 1 × 106 monocytes were incubated with CRLP (30 μg cholesterol/ml) (or a similar volume of control preparation), and incubated at 37 °C for 24 h. Cells were adhered to microscope slides by cytospin (Shandon, ThermoFisher Basingstoke, UK), and stained with Oil Red O as described previously [14]. Images were captured using a microscope mounted Canon digital camera and the extent of staining analysed Image J analysis software

(NIH). Monocytes were loaded with dihydrorhodamine-1,2,3 (final concentration 100 μM) for 10 min at room temperature and seeded Vorinostat chemical structure onto white opaque 96 well tissue culture plates (2.5 × 104 labelled monocytes/well). Pharmacological inhibitors were added for 10 min at see more 37 °C prior to addition of CRLP (7.5–30 μg/ml cholesterol) or a similar volume of control preparation. Plates were incubated

at 37 °C for up to 24 h in 5% CO2 and fluorescence was measured, using a Wallac1410 fluorescent microtitre plate reader (Perkin Elmer, Beaconsfield, UK). Monocytes were seeded at 5 × 105 cells/well in 24 well tissue culture plates and CRLP (30 μg/ml cholesterol) or a similar volume of control preparation was added. Cells were exposed to pharmacological inhibitors for 10 min at 37 °C before addition of CRLP. After incubation at 37 °C for 6 or 24 h, cells

were pelleted and the supernatants collected, snap frozen and stored Sorafenib purchase at −80 °C until analysis using ELISA Duoset assay kits according to the manufacturer’s instructions (R&D Systems, Oxford, UK). Monocytes were seeded at in 24 well tissue culture plates (5 × 105 cells/well) and exposed to CRLP (30 μg cholesterol/ml) or a similar volume of control preparation for 24 h, then transferred to the upper chamber of Transwell plates in conditioned medium. RPMI supplemented with 10% FBS (600 μl) was placed in the lower Transwell chambers and recombinant human MCP-1 (CCL2) (10 ng/ml; R&D Systems) was added to lower and/or upper chambers. The plates were incubated for 4 h and the number of cells that had migrated into the lower chamber after this time were counted by flow cytometry (Beckman Coulter, Oxford UK). Two way ANOVA followed by Bonferroni’s multiple comparison test was used to analyse ROS production and the effects of pharmacological inhibitors on cytokine production, and one way ANOVA followed by the Tukey Kramer multiple comparison test was used for all other data, except where indicated otherwise. Incubation of isolated monocytes with CRLP for 24 h resulted in increased intracellular accumulation of lipid as assessed by Oil Red O staining (Figure 1A).

Microcontact imprinting method has been used for various proteins

Microcontact imprinting method has been used for various proteins [23], [24], [25], [26] and [27]. BSA (bovine serum albumin) is a protein with the molecular weight of 66.5 kDa and it has many uses in biomedical applications and enzymatic reactions. It is used to prevent adhesion of enzymes during applications [28]. It is a generally used protein reagent in protein assays, like Bradford assay, to measure the concentration of a protein in solution. Furthermore, BSA has a structural homology with HSA (human serum albumin) [29]. Due to this, BSA is frequently studied as a model protein instead of HSA. Moreover, BSA is a commonly used target to analyze when designing new immunochemical

assays. Determination of micro-quantities of BSA is possible with methods like radioimmunoassay (RIA) or enzyme-linked immunosorbent assay Buparlisib cell line (ELISA) [30]. There are also some determinations like FT-IR spectroscopy, polarographic and fluorimetric measurements used for BSA detection [28], [31] and [32]. Some of the methods require a labelled reagent like a radioisotope or enzyme labeled antibody/antigen [30]. Some of them are really expensive and need time-consuming, complex procedures. Low selectivity

and sensitivity is the another drawback IDO inhibitor of these methods [33]. Direct, label-free, fast and sensitive measurement of various analytes with biosensors has attracted considerable interest [34]. Highly sensitive biosensor concepts make it possible to assay biomacromolecules at concentrations below the limit of detection of conventional methods [35]. Capacitive biosensors are the electrochemical sensors that measure changes in the dielectric properties PAK5 when an analyte interacts with a biorecognition element on the sensor surface, causing a decrease in the capacitance [36], [37], [38], [39], [40] and [41]. Capacitive biosensors have been used for the detection of various analytes like antigens, antibodies,

proteins and heavy metal ions [42], [43], [44], [45], [46] and [47]. These types of biosensors have a lot of advantages like inherent rapidity, high sensitivity, simplicity, low cost, easy manipulation and real-time measurement without labeling. In the study reported here, a capacitive biosensor with an automated-flow injection system was used for BSA detection. BSA is most commonly used model protein in the macromolecular imprinting studies. However, to our knowledge, this is the first microcontact-BSA imprinting study for the detection of BSA with the capacitive biosensor. Microcontact imprinting method was applied for the imprinting of BSA onto the pre-modified gold electrode surface. After modification of the gold electrode surface with poly-tyramine and acryloyl chloride, the protein stamp was brought together with a mixture of monomer and cross-linker in contact with the electrode. Thus, the microcontact BSA imprints were introduced to the electrode surface via UV-polymerization.

Inflammation is a main factor in the initiation, progression, and

Inflammation is a main factor in the initiation, progression, and acute complications of an atherosclerotic plaque [2]. Resveratrol has shown significant cardiovascular protective effects [3] in models of myocardial injury [4] and [5], systemic and pulmonary hypertension [6], and type 2 diabetes [7]. Several cardioprotective mechanisms of resveratrol, including antioxidant, anti-inflammatory, and anti-fibrotic actions, have been identified [8]. The low in vivo bioavailability caused by rapid resveratrol metabolism and elimination, its major disadvantages, limits the results for patient studies [9]. Boron

is a bioactive element for humans and boron-containing compounds present different biological activities [10]. Calcium fructoborate (CF) is INCB024360 a complex of calcium, fructose, and boron found naturally in fresh and dried fruits, vegetables,

herbs, and wine [11] and [12]. In previous studies, the effect of CF on human polymorphonuclear neutrophils and macrophages, which play a central role in the inflammatory response, has Omipalisib purchase been investigated [13] and [14]. Two very recent studies have provided important information on the possible molecular anti-inflammatory activity of CF in the treatment of osteoarthritis [15] and [16]. The purpose of this controlled pilot study was to assess the short-term synergistic effect of resveratrol in combination with CF on the clinical and biological statuses of subjects with stable angina pectoris. The combination of these two substances was based on the fact that CF acts as a stabilizer for resveratrol degradation in the digestive tract [17]. Furthermore, CF might present a positive synergism together

with resveratrol, increasing the anti-inflammatory properties of the former and the biological efficacy of the latter as an antioxidant agent. The study was randomized, double-blinded, active-controlled, Amine dehydrogenase and paralleled with three groups of subjects who received the test drugs and one control group of subjects who were not randomized. This single-center trial was approved by the institutional ethics committee of the Craiova Cardiology Center (Craiova, Romania) according to decision no. 400 in February 2010. The trial also was in accord with the Declaration of Helsinki of 1975, which was last reviewed in 2008. Placebo was not admitted by the hospital bioethics commission owing to ethical considerations. Nevertheless, this trial had a control group with subjects who fulfilled inclusion criteria, but they received only their usual medical care and treatment, without any test materials, during the clinical trial. The number of total enrolled subjects was 166 (Fig. 1). Of 116 subjects who met the inclusion criteria, 87 were included in the intention-to-treat analysis, divided into three groups (29 subjects in each group), and all completed the entire protocol.

Figure 4b shows the spectra of the chlorophyll-specific coefficie

Figure 4b shows the spectra of the chlorophyll-specific coefficient aph*(chla)(λ) for all the samples recorded as well as the average value, and the average

± SD. The variability in average aph*(chla) across all wavelengths lies within the CV range from about 29% to 94% (see also row 6 of Table 2). The smallest values of CV (29%) is reached at 675 nm, i.e. in the vicinity of the ‘red’ peak of absorption by phytoplankton pigments (the respective average value of aph*(chla) (675) is 0.0228 m2 mg−1). Throughout the range of light wavelengths between 440 and 600 nm, CV values also remain relatively small (not exceeding 40%). The presented average aph*(chla) spectra can be compared with the average spectra reported for oceanic waters by Bricaud et al. (1998) (see the dotted lines in Figure 4b representing different aph*(chla) spectra calculated Doxorubicin for four different values of Chl a   – 0.3, 1, 3 and 10 mg m−3). Our average

aph  *(chl a) spectrum is similar in shape to the two given by Bricaud et al. (1998) for Chl a   values of 3 and 10 mg m−3, but regardless of this similarity, the absolute values of our average spectrum are distinctively higher (we recall that in our study, the values of Chl a   changed over a range from less than 0.4 to more than 70 mg m−3 with an average value of about 7.6 mg m−3). Examples of best-fit power functions between aph  (440) find more and Chl a  , and aph  (675) and Chl a  , found for our Baltic data are given in Table 3. The relationship between aph  (675) and Chl a   is also plotted in Figure 5d. Compared with the similar power function fit of

aph   vs. Chl a   for oceanic waters reported by Bricaud et al. (1998) (see the dotted line in Figure 5d representing the equation for the adjacent wavelength of 674 nm: aph  (674) = 0.0182(Chl a  )0.813), the power function fit obtained in the present work shows a similar value of the power, but the value of the constant C  1 is about 50% higher. This again suggests that on average the efficiency of light Montelukast Sodium absorption (this time absorption by phytoplankton pigments alone) per unit of chlorophyll a   in our southern Baltic Sea samples is higher when compared with average oceanic results. As we said earlier, since we cannot directly compare PSDs for our Baltic samples with the size distributions for oceanic samples reported by Bricaud et al. (1998), we can only speculate about the reasons for such differences in the chlorophyll-specific absorption coefficient. Interestingly, Babin et al. (2003b) reported a qualitatively similar feature – distinctively higher aph*(chla) values for at least for some parts of the visible light spectrum for their Baltic Sea samples compared with averaged oceanic results (see the spectrum and spread of data points representing Baltic samples in their original Figures 6c and 7). Unfortunately, apart from these figures, Babin et al.