Remote sensing provides an attractive alternative We discuss the

Remote sensing provides an attractive alternative. We discuss the C59 Wnt research buy range of different sensors that are available and the differing physical manifestations of their interactions with the ocean surface. We then present existing algorithms by which the most important geophysical variables can be estimated from remote sensing measurements. Future directions and opportunities will depend on expected developments in sensors and platforms and on improving processing algorithms, including data assimilation formalisms.”
“Birt-Hogg-Dube syndrome (BHD) is an autosomal dominant disorder associated

with a germline mutation of folliculin (FLCN). The affected families are at a high risk for developing multiple renal cell carcinomas (RCC). Little is known about the immunostaining patterns of mutant FLCN-associated RCCs. We investigated 32 RCCs obtained from 17 BHD patients. The studied tumors included chromophobe RCCs (n LY2603618 concentration = 15), hybrid oncocytic/chromophobe tumors (HOCT) (n = 14) and clear cell

RCCs (n = 3). Almost all chromophobe RCCs and HOCTs revealed positive staining for S100A1, Ksp-cadherin and CD82. They stained either focally or diffusely for CK7, and were negative for CA-IX. All clear cell RCCs were positively stained for CA-IX and negative for CK7. These data confirmed that mutant FLCN-associated oncocytic and clear cell RCCs exhibited generally similar immunostaining patterns compared to their sporadic counterparts. Frequent positive staining for S100A1, Ksp-cadherin and CD82 in chromophobe RCCs and HOCTs indicated that these two types were relatively similar rather than distinctively different in their patterns of immunoreactivity. Characteristic peri-nuclear halos and

polygonal cells with clear cytoplasm, which often misleads pathologists into the diagnosis of clear cell RCC, should be carefully examined using an immunohistochemical panel selleck chemical including CA-IX, Ksp-cadherin, CD82 and CK7.”
“The present study describes the development of SYBR Green based real-time polymerase chain reaction (real-time PCR) for detection and quantitation of canine parvovirus type 2 (CPV 2) in faecal samples of dogs. In this assay, the primers were designed and custom-synthesized based on nucleotide sequence of VP2 gene of CPV 2. A standard curve was plotted using 10-fold serial dilution of standard plasmid DNA and Ct value. The standard curve was found to be linear over a 10(-7) dilution. The real-time PCR results were expressed as the number of DNA copies of CPV 2 per mg of faecal samples and showed range of 1.0 x 10(3) to 7.0 x 10(9) copies of viral DNA per mg of stool samples. The analytical sensitivity of the SYBR Green based real-time PCR was shown to be equivalent to 10 copies.

Two randomized, 6-week, double-blind cross-over trials compared t

Two randomized, 6-week, double-blind cross-over trials compared the lipid-modifying efficacy of ezetimibe/atorvastatin 10/20mg (n=353) or 10/40mg (n=280) vs. separate co-administration of ezetimibe 10mg plus atorvastatin 20mg (n=346) or 40mg (n=280), respectively, in hypercholesterolemic patients. Percent changes from baseline in LDL-C (primary endpoint) and other lipids (secondary endpoints) were assessed by analysis of covariance; triglycerides were evaluated by longitudinal-data analysis. Expected differences between FDC and the corresponding

co-administered doses were predicted from a dose-response relationship model; sample size was BMS-754807 estimated given the expected difference and equivalence margins (+/- 4%). LDL-C-lowering equivalence was based on 97.5% expanded confidence intervals (CI) for the difference

contained within the margins; equivalence margins for other lipids were not prespecified. Ezetimibe/atorvastatin Integrin inhibitor FDC 10/20mg was equivalent to co-administered ezetimibe+atorvastatin 20mg in reducing LDL-C levels (54.0% vs. 53.8%) as was FDC 10/40mg and ezetimibe+atorvastatin 40mg (58.9% vs. 58.7%), as predicted by the model. Changes in other lipids were consistent with equivalence (97.5% expanded CIs smaller than +/- 3%, included 0); triglyceride changes varied more. All treatments were generally well tolerated. Hypercholesterolemic patients administered ezetimibe/atorvastatin

10/20 and 10/40mg FDC had equivalent LDL-C lowering. This FDC formulation proved to be an efficacious and generally well-tolerated lipid-lowering therapy.”
“Microbiomes associated with multicellular organisms influence the disease susceptibility of hosts. The potential exists for such bacteria to protect wildlife from infectious diseases, particularly in the case of the globally distributed and highly virulent fungal pathogen Batrachochytrium dendrobatidis of the global panzootic lineage (B. dendrobatidis GPL), responsible for mass extinctions and population declines of amphibians. B. dendrobatidis GPL exhibits wide genotypic and virulence variation, and the ability of candidate probiotics to restrict growth across B. dendrobatidis selleck compound isolates has not previously been considered. Here we show that only a small proportion of candidate probiotics exhibited broad-spectrum inhibition across B. dendrobatidis GPL isolates. Moreover, some bacterial genera showed significantly greater inhibition than others, but overall, genus and species were not particularly reliable predictors of inhibitory capabilities. These findings indicate that bacterial consortia are likely to offer a more stable and effective approach to probiotics, particularly if related bacteria are selected from genera with greater antimicrobial capabilities.

For the 3D AFI method, the addition of flow-compensated gradients

For the 3D AFI method, the addition of flow-compensated gradients for diffusion damping reduced the level of physiological artifacts and improved spoiling of transverse coherences. Correction of susceptibility-induced artifacts alleviated image distortions and improved the accuracy of the 3D EPI imaging method. For the 2D STEAM method, averaging over multiple acquisitions reduced the impact of physiological noise and a new calibration

method enhanced the accuracy of the B(1)(+) maps. After optimization, all methods yielded low noise B(1)(+) maps (below 2 percentage units), of the nominal flip angle value (p.u.) with a systematic bias less than 5 p.u. units. Full brain coverage AZD1208 was obtained in less than 5 min. The 3D AFI method required minimal postprocessing and showed little sensitivity to off-resonance and physiological effects. The 3D EPI method showed the highest level of reproducibility. The 20 STEAM method was the most time-efficient technique. Magn Reson Med 64:229-238, 2010. (C) 2010 Wiley-Liss, Inc.”
“Background:Restricted

diffusion of water molecules on diffusion-weighted magnetic resonance imaging most commonly associated with acute stroke, has also been described in brain abscess. It has been reported in only one case of sub-retinal abscess.Methods:Review of two cases.Results:Two cases of visual loss from subretinal abscess APR-246 manufacturer had restricted diffusion in the region of the abscess. In the first case, DWI revealed restricted diffusion in a white mass visible in the posterior retina. In the second case, restricted diffusion was present in an anterior retinal mass invisible by ophthalmoscopy and ultrasound. In combination of restricted diffusion in the cerebrum consistent with septic emboli, these imaging abnormalities allowed earlier treatment that either preserved vision or prevented enucleation.”
“Background: Considering the dramatic increasing rate of diabetes and consequently its related complications, most importantly diabetic peripheral neuropathy (DPN), challenges regarding proper treatment

of DPN and its effect on the quality-of-life and care of diabetic patients, the aim of this current study is to evaluate the effect of intradermal botulinum toxin type A (BTX-A) injections PLX4032 on pain symptoms of patients with diabetic neuropathic pain. Materials and Methods: In this randomized double-blind placebo-controlled clinical trial study, diabetic patients aged smaller than 70 years with neuropathic pain in both feet were enrolled. Diabetic neuropathy (DN) in selected patients was diagnosed using DN4 questionnaire and nerve conduction velocity examinations. They randomized in two intervention (BTX-A injection/100 unit, N = 20) and placebo groups (normal saline injection, N = 20). The outcome of injection on diabetic neuropathic pain was assessed using neuropathy pain scale (NPS) and visual analog scale (VAS) score and compared in two studied groups.

Several genes which showed a restricted expression to lung tissue

Several genes which showed a restricted expression to lung tissue were identified. Using correlations between gene expression values across all strains, we defined and extended memberships of several important molecular networks in the lung. Furthermore, we were able to extract

signatures of immune cell subpopulations and characterize co-variation and shared genetic modulation. Known QTL regions for respiratory infection susceptibility were investigated and several cis-eQTL genes were identified. Numerous cis- and trans-regulated transcripts and chromosomal intervals with strong regulatory activity were mapped. The Cyp1a1 P450 transcript had a strong trans-acting eQTL (LOD 11.8) on Chr 12 at 36 +/- 1 Mb. This interval contains the transcription factor Ahr that GW-572016 cell line has a critical mis-sense allele in the DBA/2J haplotype and evidently modulates transcriptional activation by AhR.\n\nConclusions: Large-scale gene expression

analyses in genetic reference populations revealed lung-specific and immune-cell gene expression profiles and suggested specific gene regulatory DNA-PK inhibitor interactions.”
“Background: Acquired immunodeficiency syndrome (AIDS), which is caused by the human immunodeficiency virus (HIV), is an immunosuppressive disease that results in life-threatening opportunistic infections. The general problems in current therapy include the constant emergence of drug-resistant HIV strains, adverse side effects and the unavailability of treatments in developing countries. Natural products from herbs with the abilities to inhibit HIV-1 life cycle at different stages, have served as excellent sources of new anti-HIV-1 drugs. In this study, we aimed to investigate the anti-HIV-1 activity of aqueous Go 6983 molecular weight dandelion extract.\n\nMethods: The pseudotyped HIV-1 virus has been utilized

to explore the anti-HIV-1 activity of dandelion, the level of HIV-1 replication was assessed by the percentage of GFP positive cells. The inhibitory effect of the dandelion extract on reverse transcriptase activity was assessed by the reverse transcriptase assay kit.\n\nResults: Compared to control values obtained from cells infected without treatment, the level of HIV-1 replication and reverse transcriptase activity were decreased in a dose-dependent manner. The data suggest that dandelion extract has a potent inhibitory activity against HIV-1 replication and reverse transcriptase activity. The identification of HIV-1 antiviral compounds from Taraxacum officinale should be pursued.\n\nConclusions: The dandelion extract showed strong activity against HIV-1 RT and inhibited both the HIV-1 vector and the hybrid-MoMuLV/MoMuSV retrovirus replication. These findings provide additional support for the potential therapeutic efficacy of Taraxacum officinale. Extracts from this plant may be regarded as another starting point for the development of an antiretroviral therapy with fewer side effects.


“A statistical analysis and computational algorithm for co


“A statistical analysis and computational algorithm for comparing pairs of tool marks via profilometry data is described. Empirical validation of the method is established through experiments based on tool marks made at selected fixed angles from 50 sequentially manufactured screwdriver tips. Results

obtained from three different comparison scenarios are presented and are in agreement with experiential knowledge Erastin possessed by practicing examiners. Further comparisons between scores produced by the algorithm and visual assessments of the same tool mark pairs by professional tool mark examiners in a blind study in general show good agreement between the algorithm and human experts. In specific instances where the algorithm had difficulty in assessing a particular comparison pair, results obtained during the collaborative study with professional examiners suggest ways in which algorithm Repotrectinib in vivo performance may be improved. It is concluded that the addition of contextual information

when inputting data into the algorithm should result in better performance.”
“Ascorbate (AsA) is an important antioxidant and an enzyme cofactor involved in various metabolic pathways. In this study, we investigated the effects of estrogen (ES)-inducible transient expression of genes encoding enzymes involved in the d-mannose/l-galactose (d-Man/l-Gal) pathway for plant AsA biosynthesis on AsA levels under light and dark conditions. No significant difference was observed in AsA levels between Arabidopsis plants transiently expressing phosphomannose isomerase (PMI1), GDP-d-Man pyrophosphorylase (GMP/VTC1), GDP-Man-3 ‘,5 ‘-epimerase (GME), and l-Gal 1-phosphate phosphatase (GPP/VTC4), but AsA levels in the plants transiently expressing GDP-l-Gal phosphorylase (GGP/VTC2) Fedratinib manufacturer were 2.5-fold higher than those in control plants 7 d after ES treatment. The increase in AsA levels under continuous light conditions and the decrease

in AsA levels under dark conditions were enhanced and suppressed, respectively, in the ES-treated plants. These results suggest that GGP/VTC2 acts as a rate-limiting step regulating AsA biosynthesis in response to light and dark conditions.”
“Purpose: To report a case of delayed spontaneous Descemet membrane (DM) tear after big-bubble Deep Anterior Lamellar Keratoplasty (DALK). Methods: Uneventful big-bubble DALK was performed on a 29-year-old patient with advanced keratoconus. No injury to DM was noted intraoperatively and in the first postoperative week. On examination after 1 month, the patient presented with tear and partial detachment of Descemet membrane (DMD). Results: Circumscribed eccentric stromal edema, but not DMD, partially resolved after a 3-month observational period.

We propose that Mtb Eis initiates the inhibition of JNK-dependent

We propose that Mtb Eis initiates the inhibition of JNK-dependent autophagy, phagosome maturation, and ROS generation by acetylating DUSP16/MKP-7. Our work thus provides insight into the mechanism of suppressing host

immune responses and enhancing mycobacterial survival within macrophages by Mtb Eis.”
“Moving a set dinner table often takes two people, and doing so without spilling the glasses requires the close coordination of the two agents’ actions. It has been argued that the mirror neuron system may be the key neural locus of such coordination. Instead, here we show that such coordination recruits two separable sets of areas: one that could translate between motor and visual codes and one that could integrate these information to BMS-754807 supplier achieve common goals. The former includes regions of the putative mirror neuron system, the latter, regions

Cilengitide cell line of the prefrontal, posterior parietal and temporal lobe adjacent to the putative mirror neuron system. Both networks were more active while participants cooperated with a human agent, responding to their actions, compared to a computer that did not, evidencing their social dimension. This finding shows that although the putative mirror neuron system can play a critical role in joint actions by translating both agents’ actions into a common code, the flexible remapping of our own actions with those of others required during joint actions seems to be performed outside of the putative mirror neuron system. (C) 2009 Elsevier Inc. All rights reserved.”
“Background. Diabetic nephropathy (DN) is a common microvascular complication of diabetes. In this study, we aimed to explore both primary effects of single-locus

and multilocus interactions to test the hypothesis that the type 2 diabetes (T2D) genes may contribute to the aetiology of DN in T2D independently and/or through complex interactions in a Taiwanese population with T2D.\n\nMethods. We genotyped six single nucleotide GSK2245840 datasheet polymorphisms (SNPs) for five common T2D genes including adiponectin, C1Q and collagen domain containing (ADIPOQ), ectonucleotide pyrophosphatase/ phosphodiesterase 1 (ENPP1), growth hormone secretagogue receptor (GRSR), peroxisome proliferator-activated receptor gamma (PPAR gamma) and transcription factor 7-like 2 (TCF7L2). There were 216 T2D patients diagnosed with DN and 178 age-similar T2D without DN (control) subjects. To investigate gene-gene interactions, we employed both generalized multifactor dimensionality reduction (GMDR) method and logistic regression models.\n\nResults. Single-locus analyses showed significant main effects of ENPP1 (P = 0.0032; adjusted OR = 1.85; 95% CI = 1.17-2.92) on the risk of DN in T2D. Furthermore, a potential gene-gene interaction involving ENPP1 and GHSR was suggested in the best two-locus GMDR model (P = 0.021). The significant three-locus GMDR model (P < 0.

) seeds was determined by capillary GC and GC/MS Thirty-two fatt

) seeds was determined by capillary GC and GC/MS. Thirty-two fatty acids (99.9%) have been identified in the fixed oil. The major fatty acids were linoleic acid (50.2%), oleic acid (19.9%), margaric acid (10.3%), cis-11,14-eicosadienoic acid (7.7%) and stearic acid (2.5%). The effect of black cumin oil on in vitro percutaneous absorption of the model lipophilic Omipalisib in vitro drug carvedilol was investigated using excised rat abdominal skin. Transdermal flux, permeability coefficient and enhancement factor were calculated for different concentrations of oil in isopropyl alcohol. Black cumin oil (5% v/v)

exhibited the highest enhancement in permeation. The increase in the permeability of the drug is due to increased drug diffusivity through the stratum corneum under the influence of black cumin oil. A higher content of linoleic acid (and other unsaturated fatty acids) in the oil has been postulated to be responsible for the enhancement

of in vitro percutaneous absorption of the drug.”
“Most PLX3397 ic50 of the arborescent Croton species in the New World were treated by Webster as belonging either to C. sect. Cylostigma Griseb. or C. sect. Luntia (Neck. ex Raf.) G.L. Webster. The circumscription of C. sect. Cyclostigma has been treated recently. In this paper we focus on C. sect. Luntia, which was subdivided by Webster into two subsections, C. subsect. Matourenses and subsect. Cuneati. Using chloroplast trnL-F and nuclear ITS DNA sequence

data, morphology and a broader sampling of additional Croton lineages, we found that the two subsections are not closely related and form distinct monophyletic groups. Substantial morphological differences support the molecular data. A taxonomic recircumscription of the two subsections, elevated to sectional level, is proposed. A key and taxonomic revision, with new synonyms, is provided for C. sect. Cuneati; together with the description of a new Histone Methyltransf inhibitor species from the coastal mountains of Venezuela, Croton domatifer. The new species is the only one in the genus known to possess leaf domatia. We infer that species in the Cuneati clade have lost the typical Croton caruncle, and some of them have evolved a different, specialized type of aril. We hypothesize that the arillate seeds are an adaptation to dispersal by fish in the Orinoco and Amazon river basins.”
“We investigated physiological parameters (elemental and biochemical composition, metabolic rates, feeding activity and growth) of adult Antarctic krill in the Lazarev Sea in late spring (December), mid autumn (April) and mid winter (July and August) to evaluate proposed hypotheses of overwintering mechanisms.


“Background: Hepatitis

B virus-associated glomerul


“Background: Hepatitis

B virus-associated glomerulonephritis (HBV-GN) mainly occurs in children. Patients with HBV-GN are frequently positive for serum HBV surface antigen (HBsAg), but they are rarely negative. Objective: To explore the clinical and pathological characteristics of patients with HBV-GN who are serum HBsAg negative. Study design: Five children with HBV-GN who are negative for HBsAg were included in this study. Their clinical and pathological characteristics were collected and analyzed. Results: All 5 children presented with different levels of proteinuria and microscopic hematuria. Renal biopsies showed membranous nephropathy accompanied by HBsAg and/or HBcAg deposits in glomeruli in all of the children. Steroids and/or other immunosuppressants were administered in all cases without antiviral therapy during the early stages of treatment. Two children achieved complete remission STA-9090 manufacturer but relapsed after the drugs were tapered down. The other 3 children were initially non-responsive but achieved remission after lamivudine was added. Conclusions: Treatment of HBsAg-negative HBV-GN patients with immunosuppressants alone could not achieve satisfactory LY3023414 supplier effects. Antiviral

treatment is effective and may be necessary in this type of patient. (C) 2015 Elsevier B.V. All rights reserved.”
“Effects of superficial gas velocity and top clearance on gas holdup, liquid circulation velocity, selleck chemicals mixing time, and mass transfer coefficient are investigated in a new airlift loop photobioreactor (PBR), and empirical models for its rational control and scale-up are proposed. In addition, the impact of top clearance on hydrodynamics, especially on the gas holdup in the internal airlift loop reactor, is clarified; a novel volume expansion technique is developed to determine the low gas holdup in the PBR. Moreover, a model strain of Chlorella vulgaris is cultivated in the PBR and the volumetric power is analyzed with a classic model,

and then the aeration is optimized. It shows that the designed PBR, a cost-effective reactor, is promising for the mass cultivation of microalgae. (C) 2015 Elsevier Ltd. All rights reserved.”
“Bone substitute materials allowing trans-scaffold migration and in-scaffold survival of human bone-derived cells are mandatory for development of cell-engineered permanent implants to repair bone defects. In this study, we evaluated the influence on human bone-derived cells of the material composition and microstructure of foam scaffolds of calcium aluminate. The scaffolds were prepared using a direct foaming method allowing wide-range tailoring of the microstructure for pore size and pore openings. Human fetal osteoblasts (osteo-progenitors) attached to the scaffolds, migrated across the entire bioceramic depending on the scaffold pore size, colonized, and survived in the porous material for at least 6 weeks.

e , higher’ school-level SES) There was also an indication of mo

e., higher’ school-level SES). There was also an indication of moderation of the shared environment; there were greater shared environmental influences on reading

comprehension at higher school-level Selleck Anlotinib SES.\n\nConclusionsThe results supported the bioecological model; greater genetic variance was found in school environments in which student populations experienced less poverty. In general, higher’ school-level SES allowed genetic and probably shared environmental variance to contribute as sources of individual differences in reading comprehension outcomes. Poverty suppresses these influences.”
“Purpose This study investigated changes in depressive symptoms after the implementation of a universal screening for depression and subsequent care support. Methods A cluster-randomized study design used 10 subdistricts (2,400 inhabitants aged 40-64 years) in northern Japan randomly assigned in a 2:3 ratio to intervention and control conditions.

All 900 residents aged 40-64 in the intervention districts were invited to participate in a 2-year depressive screening program, with a participation rate MI-503 of 49.2 %. A 4-year ongoing education program occurred in both intervention and control districts. The Center for Epidemiologic Studies Depression Scale (CES-D) was used to assess depressive symptomatology. Repeated cross-sectional samples were surveyed before (n = 1,516, response rate 63.6 %) and after (n = 1,596, 66.4 %) intervention, and the data, clustered according to district, were analyzed at the individual level using a mixed-effects model. Results Significant changes in mean scores between baseline and 5-year follow-up in the intervention group were observed in the Depressive Affect, Somatic Symptoms, and Interpersonal Problems subscales. learn more The difference between the changes over time in the two groups was significant for the three subscales and marginally for the CES-D total scale, but not for the Positive Affect

subscale. Conclusions Universal depression screening and subsequent support can be effective in preventing general depressive symptoms, but may not influence psychological well-being, among middle-aged adults in a community setting.”
“Chitosan-based nanoparticles (chiNPs) are considered to be potentially good carriers for the sustained intracellular delivery of specific molecules. However, scarce attention has been paid to the long-lasting permanence of these NPs in the intracellular milieu, as well as to their intracellular fate (i.e., distribution, interaction with cell organelles, and degradation) in the long term. In the present study, the presence and subcellular location of FITC-labelled chiNPs were monitored in HeLa cells up to 14 days post-administration using multicolor-fluorescence confocal microscopy and diaminobenzidine photo-oxidation at transmission electron microscopy.

But concern lingers that DAs might introduce cognitive biases \

But concern lingers that DAs might introduce cognitive biases.\n\nMethods: We recruited 663 women at high risk of breast cancer and presented them with a DA designed to experimentally test potential methods of identifying and reducing cognitive biases that could influence this decision, by varying specific Compound C in vitro aspects of the DA across participants in a factorial design.\n\nResults: Participants were susceptible to a cognitive bias – an order effect – such that those who learned first about the risks of tamoxifen thought more favorably of the drug than women who learned first about

the benefits. This order effect was eliminated among women who received additional information about competing health risks.\n\nConclusion: We discovered that the order of risk/benefit information influenced women’s perceptions of tamoxifen. This bias was eliminated by providing

contextual information about competing health risks. Practice implications: We have demonstrated the feasibility of using factorial experimental designs to test whether DAs introduce cognitive biases, and whether specific elements of DAs can reduce such biases. Published by Elsevier Ireland Ltd.”
“The aim of this study was to evaluate the effects of an analogue of lacidipine, CZ454 in in vitro and in vivo. The isometric tension of Sprague-Dawley rat arterial ring segments was recorded by a myography system. Intracellular calcium of vascular smooth muscle was determined by the confocal laser microscopy. Blood pressure of spontaneously hypertensive rats was measured using AZD5363 molecular weight a tail-cuff blood pressure system. The results showed that CZ454 (10(-9)-10(-6)mol/L) relaxed

the mesenteric artery selleck contracted by high K+ concentration-dependently, which was not affected by removal of the endothelium. CZ454 treatment shifted the concentration-contractile curves induced by phenylephrine, U46619, KCl and CaCl2 to the right with the decreased E-max. CZ454 was more potent in the coronary and basilar artery than in the mesenteric artery. CZ454 did not reduce phenylephrine-induced vasoconstriction: however, it did inhibit the contraction caused by addition of CaCl2 and did not change caffeine-induced contraction in the mesenteric artery in Ca2+-free solution. CZ454 decreased the vasoconstriction induced by Bay K 8644 in the presence of 60mmol/L K+ CZ454 1.0 mg/kg administered by gavage lowered the systolic pressure and diastolic pressure by 20% and 17%, respectively. It was concluded that CZ454 lowers blood pressure and relaxes arteries with higher potency in coronary and basilar artery and that the vasodilation may involve inhibition of calcium influx.”
“Objective: The link between impulsive personality traits and alcohol use disorders (AUDs) is well established. No studies, however, have investigated whether receipt of help for AUDs predicts change in impulsivity or whether such change is associated with relevant outcomes such as legal problems.