The products of the two genes formed a complex with efflux transport activity specific for UDP-glucose, of which exogenous addition protected root growth under Al stress. Protein activity of Al-tolerance genes BnALMT1 and BnALMT2 in Brassica was tested in tobacco

cells and Xenopus oocytes and showed that they conferred malate efflux, and transgenic tobacco cells had enhanced tolerance to Al toxicity [143]. The rapid development of molecular markers and QTL mapping of Al tolerance permits MAS for Al tolerance in breeding programs. Traditional click here breeding has benefited from conventional selection based on phenotyping; however, phenotypic selection is reportedly difficult, inefficient and laborious due to its dependence on specific environments [144]. MAS is based on associations between molecular markers and superior alleles of genetic traits of interest. After QTL are validated, tightly-linked markers can be used to detect, transfer and check details accumulate desirable genome regions into superior genotypes, a process that is much faster than phenotypic selection. The major advantages of MAS compared to conventional phenotypic selection are cost-effectiveness, simplicity of selection, time-saving and screening precision [145]. Different types of markers have been developed to trace interesting genes or loci. As discussed in

a previous section, molecular markers including RFLP, AFLP, RAPD, SSR, DArT and SNP have been developed and used in Al-tolerance studies. These have proved efficient in MAS in breeding programs. With increasing Farnesyltransferase numbers of genes for Al tolerance being identified and sequenced in plants, PCR-based gene-specific markers developed from gene sequencing are preferred in MAS for their easy identification, high polymorphism and good reproducibility [146]. In wheat, Raman et al. [158] developed SSR markers, ALMT1-SSR3a and ALMT1-SSR3b and a CAPS marker from the repetitive InDels and substitution region of the TaALMT1 gene. These PCR-based markers co-segregating with the tolerance locus should be efficient tools for MAS [147]. In barley, one gene-specific marker, HvMATE-21indel,

was developed from the tolerance gene HvMATE. The marker increased the explained phenotypic variation compared with the other SSR markers. It can also be used for selecting the tolerance gene from multiple tolerance sources [148]. With additional and different types of molecular markers being developed for Al tolerance, breeding programs could be accelerated by using these markers in MAS [78]. Transgenic methods are very efficient for validating gene function in Al-tolerance studies. The first report on a transgenic approach to increasing Al tolerance in plants was in 1997 when De La Fuente et al. [149] reported that an overexpressed citrate synthase gene enhanced citrate efflux and led to improved root Al tolerance in transgenic tobacco.

Fifty micrograms of total protein was separated on sodium dodecyl sulfate–polyacrylamide gels and transferred onto nitrocellulose membranes click here (Merck Millipore, Billerica, MA). The blots were probed with antibodies against

YAP, phospho-YAP (pYAP), and glyceraldehyde 3-phosphate dehydrogenase (GAPDH) (Cell Signaling Technology) as well as TEAD1 (Santa Cruz Biotechnology, Dallas, TX). Secondary, HRP-coupled antibodies directed against rabbit or mouse IgG, respectively, were purchased from Cell Signaling Technology. Detection was performed as previously described [13]. A total of 100 ng of total RNA, isolated using the RNeasy Kit (Qiagen, Hilden, Germany) following the standard procedure as recommended by the manufacturer, was subjected to a single round of in vitro transcription and biotin labeling (Illumina TotalPrep RNA Amplification Kit; Ambion, Austin, TX). The resulting complementary RNA was hybridized on HumanHT-12 v4.0 Expression BeadArrays (Illumina, San Diego, CA) according to the manufacturer’s protocols using an automated liquid handling pipeline and scanned on an iScan System. Expression data were exported as unnormalized sample and control probe profiles from the Illumina GenomeStudio software and analyzed using R/Bioconductor and limma. Data were quality weighed, background-corrected,

quantile-normalized, log-transformed, and explored for differentially expressed genes with a false discovery rate (FDR) of 0.05 using Bayesian statistics. Sorafenib molecular weight Differential regulation of signaling pathways was performed using the signaling pathway impact analysis algorithm as previously described elsewhere [14]. For real-time reverse transcription–quantitative PCR (RT-qPCR) analysis, cells were lysed and total RNA was extracted using RNeasy Mini Kit

including an on-column Amylase DNase digestion step (both Qiagen). RNA was reverse transcribed using random primers and the High Capacity cDNA Reverse Transcription Kit (Life Technologies) according to the manufacturer’s specifications. Real-time qPCR for human YAP, endothelin-1 (EDN1), EDN2, V-myc myelocytomatosis viral oncogene homolog (avian) (MYC), cadherin-6 (CDH6), cysteine-rich, angiogenic inducer, 61 (CYR61), thrombospondin-1 (THBS1), and growth arrest and DNA damage-inducible, beta was performed using the SYBR Premix Ex Taq (Tli RNase H Plus) Kit (Takara Bio Europe, Saint-Germain-en-Laye, France) on a Reaplex2 Mastercycler Real-Time PCR System (Eppendorf, Hamburg, Germany). Relative fold expression levels were determined using the 2(− ΔΔCt) method [15], with GAPDH used as a housekeeping control. TEAD1-binding sites found in 5 kb upstream of the transcription start site of the indicated genes were considered and primer pairs for qPCR measurement of immunoprecipitated promoter fragments flanking the putative binding regions were designed.

Both non-nucleoside inhibitors and nucleoside inhibitors of NS5B are currently being tested in clinical

trials, and the most advanced protease inhibitors, such as telaprevir, are linear compounds and are currently on the market; several macrocyclic compounds this website are being tested in clinical trials.11, 22, 23, 24 and 25 Monotherapy with the non-nucleoside polymerase inhibitor, HCV-796, showed less than a 0.5 log10 reduction of serum HCV RNA levels, while a combined treatment with NA808 reduced the HCV titer by 1000-fold from the initial serum levels. This effect was higher than the effect of NA808 alone and higher than the sum of NA808 and HCV-796 monotherapy effects, suggesting synergistic Z-VAD-FMK mouse antiviral efficacy. A similar effect was observed by combination of NA808 with nucleoside polymerase inhibitor, RO-9187, as shown in Figure 4B. The maximum reductions in HCV RNA are mediated by triple combinatorial treatment and the significant in vivo anti-HCV activity with combination treatment is also

observed when NA808 is combined with PEG-IFN. These observations suggest that NA808 may have synergistic antiviral activity with various classes of anti-HCV agents, regardless of their inhibition mechanism, due to the unique host enzyme-targeted mechanism of action. In addition, NA808 could be expected to show a higher barrier to the development of resistant clones. Deep-sequencing analysis showed no evidence for the development of NA808 resistance after 14 passages

in HCV replicon cells, while telaprevir treatment resulted in the selection of known protease resistance mutations (V36A, T54V, and A156T) ( Table 2). The full-genome sequence of HCV obtained see more at day 14 from HCV-infected humanized-liver mice treated with NA808 for 14 days also showed no evidence for the selection of resistance mutations, consistent with the viral load kinetics ( Figure 2B). Host enzyme inhibition might be associated with mechanism-related toxicities or side effects. Although more thorough analyses of toxicity with NA808 are warranted, NA808 did not affect host cell viability in vitro under the assay conditions used (Supplementary Figure 1A), and it was well-tolerated in vivo at the efficacious dose used. The effective plasma concentration of NA808 at trough level was approximately 1.4 nmol/L ( Table 3), around 100 times lower than rats that received 40 mg/kg NA808 at 24 hours after injection (data not shown). No NA808-related changes, including abnormalities of general conditions, body weight decreases, and macroscopic or microscopic changes were observed at this high dose in rats. Homozygous knockout mice for sptlc1 and sptlc2, subunits of SPT, were embryonic lethal, and heterozygous mice showed no phenotype. 26 Mice with conditional sptlc2 knockout showed necrotic lesions in gastrointestinal cells.

19 When oral food and ONS are impossible or inadequate, nutrition can be given as enteral tube feeds. When the gastrointestinal tract is so compromised that calorie and protein requirements cannot be fully Small Molecule Compound Library met by enteral feeding, parenteral nutrition can be used either alone or in combination with enteral nutrition. Guidelines support prompt intervention, that is, individualized nutrition therapy within 24 to 48 hours of admission.7, 16, 17 and 88 As a notable exception, a patient

near the end of life can be kept comfortable without provision of food or oral/enteral nutrition, if this strategy is mutually agreeable to patient/family and caregivers.89 Many hospitalized individuals are able to eat food, but their appetite is limited by illness. In such cases, experts recommend foods with energy-rich additives (eg maltodextrin, protein fortification), eating smaller but more frequent meals or high-energy snacks between meals, or using ONS.7 Standard commercially prepared enteral formulas are complete and balanced and contain an energy level of 1.0 kcal/mL, thus meeting the needs of many sick or injured patients who cannot

get adequate nutrition with a diet of regular food.90 Specialized commercially prepared formulas meet basic needs but also meet disease- or condition-specific needs, including 1.0 to 2.0 kcal/mL; some are formulated and flavored for use as ONS or enteral tube feeds, and others are intended only for enteral tube feeds.91 Nutrition care PD-0332991 price does not end when a patient is released from the hospital or other care center. The final step of the Nutrition Care Pathway is to supervene below and follow-up, with continuing attention to meeting nutrition needs. In fact, poor nutritional status on discharge predicts hospital readmission within 30 days.92 New focus on postdischarge nutrition planning18 is expected to help lower costly hospital readmissions,20

improve quality of life for patients,53 and 55 and in some cases even reduce risk of death.25 Effective nutrition care necessitates a postdischarge nutrition plan, and use of follow-up measures to ensure that the plan is implemented. Results of a systematic review of 6 RCTs (surgical and medical patients of older age) showed that postdischarge nutrition care with use of ONS had a positive effect on nutritional intake (energy) and nutritional status (weight) in all trials.93 The feedM.E. Global Group thus recommends continued efforts to prevent and treat malnutrition for patients who have been discharged from the hospital into long-term care centers or into the community. Attention to nutrition is fundamental to good clinical practice. Nutrition care improves patient outcomes and reduces health care costs. We, the members of the feedM.E. Global Group on Nutrition in Healthcare, call health care providers worldwide to action with “screen, intervene, and supervene.

There was a significant decrease in sCTX-1 from baseline in both treatment groups at month 1 and a significantly

greater reduction was observed with denosumab treatment compared with risedronate treatment: median (IQR) percentage change of − 77.7% (− 85.9%, − 67.6%) for denosumab and − 17.0% (− 36.8%, − 1.6%) for risedronate (p < 0.0001; Fig. 4). Median reductions in sCTX-1 at month 6 were also greater in the denosumab group compared with the risedronate group: median (IQR) percentage change of − 60.6% (− 77.0%, − 48.8%) for denosumab and GSK1120212 –22.5% (− 41.9%, 11.4%) for risedronate (p < 0.0001). The BMD mean percentage changes from baseline at month 12 by tertiles (< 0.23, ≥ 0.23 to < 0.37, and ≥ 0.37 ng/mL) of baseline sCTX-1 for each skeletal site are reported in Fig. 5. This additional analysis showed that subjects treated with denosumab, compared with risedronate, demonstrated significantly greater gains in lumbar spine BMD at month 12 at each tertile of baseline sCTX-1 (p < 0.01; Fig. 5). Significantly greater gains in total hip and femoral neck BMD were also observed among subjects in the middle and highest tertiles of baseline sCTX-1 (p < 0.01). At all sites the magnitude of the BMD gain was significantly more Selleck PR171 pronounced in the middle and highest sCTX-1 tertiles (treatment-by-sCTX-1 tertile

interaction p-values < 0.01). The post-hoc analysis showed that nearly half of the enrolled population was at higher risk for fracture: 46.4% and 45.5% of risedronate- and denosumab-treated subjects, respectively. These higher-risk subjects demonstrated BMD gains that were consistent with findings in the overall population (Fig. 6). Overall, the subject incidences of AEs were 293 subjects click here (68.3%) in the risedronate group and

269 subjects (62.7%) in the denosumab group, with the most frequently experienced AEs (≥ 4% in either treatment group [risedronate, denosumab]) being hypertension (2.6%, 4.2%), arthralgia (4.4%, 4.0%), nasopharyngitis (4.2%, 3.5%), and constipation (5.1%, 3.3%). Most of the AEs in both groups were categorized as being either mild or moderate in severity (Table 2). SAEs were reported for 8.2% of risedronate-treated subjects and 7.7% of denosumab-treated subjects. There was no evidence of clustering of SAEs within any given system organ class or high-level group term in either treatment group. SAEs reported for ≥ 2 denosumab-treated subjects were osteoarthritis, radius fracture, cerebral ischemia, cerebrovascular accident, arthralgia, and atrial fibrillation; these SAEs were each experienced by 2 (0.5%) denosumab-treated subjects. In the risedronate treatment group, the most frequently reported SAEs (2 [0.5%] subjects each) were breast cancer and coronary artery stenosis; all other SAEs were experienced at an incidence of 1 subject each. One death due to cardiac arrest was reported in a risedronate-treated subject.

Because apnea is accompanied by hypoxia and hypercapnia and pCO2 and perivascular pH are major regulatory determinants of CBF and flow velocity, changes in cerebral hemodynamics are to be expected in patients with SAS [35], [41], [42] and [61]. These theoretic considerations have been confirmed by a limited number of studies. Meyer et al. [62] performed CBF measurements

during daytime sleeping and waking states in 13 patients with narcolepsy and 7 with SAS. In the waking state, brainstem, cerebellar and bihemispheric flow were below normal in both patient groups. After sleep onset, CBF decreased further; maximum changes of regional flow values were seen in brainstem regions, indicating a critically reduced brainstem functional activity during sleep in SAS. Alterations of flow velocities during apnea-associated changes of CO2 were also reported in obstructive SAS [63]. Now that

click here several studies have shown that transcranial Doppler sonography is a useful Apoptosis Compound Library price method for long-term and on-line monitoring of dynamic changes in cerebral perfusion during sleep, researchers have begun using TCD for the assessment of perfusion changes in pathological sleep conditions. Various studies have been performed to assess cerebral flow velocity changes during nocturnal apneic episodes in patients with SAS. Siebler et al. [64] were the first to observe a cerebral flow velocity increase during nocturnal apneic phases in a patient with obstructive SAS and their findings have since been confirmed by various independent work groups in larger numbers of patients [65], [66] and [67]. Fischer et al. [34], who compared

the MFV changes in SAS patients with those of a comparable control group, observed lower MFV values in SAS patients during wakefulness, NREM sleep and REM sleep than in normals. They therefore concluded that altered cerebral perfusion occurs in SAS patients. However, a sleep stage-correlated CBF velocity assessment in SAS patients and normal control subjects determined that the course of CBF velocity changes in apneic patients during night sleep were comparable to those observed in healthy control subjects. These findings indicate that the general pattern of cerebral perfusion changes associated with sleep Niclosamide remains preserved in SAS and they contradict the hypothesis of the existence of cerebral hypoperfusion in SAS [65] and [66]. Klingelhöfer et al. [66] observed MFV increases of 19–219%, reaching a maximum in REM sleep, during apneic episodes in 6 patients with SAS (age: 34–55 years, mean age: 49 years) (Fig. 8). There was also a significant increase in blood pressure (12.5–83.1%) during apneic episodes. A multiple linear regression analysis revealed that the flow velocity increase was not only attributable to the blood pressure increase alone, but was significantly linked to apnea.

Two patients in the 60-U/kg treatment group Venetoclax order experienced 8 events that were considered to be treatment-related by the investigator; the first patient reported 3 occurrences of itchy throat and 1 occurrence of chest discomfort, and the second patient reported 1 occurrence of gastroenteritis and 3 occurrences of vomiting. Both patients were pre-medicated with H1 blockers and are continuing in the extension study. No patient withdrew from this trial due to an AE. No clinically significant laboratory test abnormalities (hematology,

serum and urinary chemistry) were noted. There were no clinically significant mean changes from baseline observed at the end of the study in any of the laboratory safety parameters. The majority of the vital sign measurements were within normal limits, none of the changes or the measurements outside of the normal limits were clinically significant. Abnormal echocardiography results at month 12 were reported for 2 patients: One patient receiving the 60-U/kg dose had mild tricuspid regurgitation and 1 patient in the 30-U/kg group diagnosed with Type 3c GD (subtype known to have a cardiac involvement) [1], [17] and [18] had a baseline echocardiography that revealed abnormal atrioventricular and mitral valves with

an insufficiency gradient of 30 mm Hg. At study end, the echocardiography results showed pulmonary hypertension with an abnormal tricuspid insufficiency gradient of 74 mm Hg, which was considered a clinically significant deterioration buy 5-FU but was deemed not related to study treatment. Two patients were found to be IgG positive for anti-taliglucerase-alfa antibodies in at least 1 post-treatment visit; however, this finding did not affect the continued improvement of GD parameters throughout the course of the study. An additional patient was found to be IgG positive at the pretreatment sample and became negative as the trial progressed; this patient also improved clinically as noted above for the other 2 patients. All positive titers were low (< 550). Assay results for neutralizing antibodies (in vitro enzymatic inhibition assay and cell-based neutralizing

assay) were negative for all 3 patients. No apparent association was noted between anti-taliglucerase antibody and safety or efficacy. This study is distinguished among studies of ERT Docetaxel datasheet for GD in that it is focused exclusively on treatment-naïve pediatric patients. This pediatric study followed the design of the pivotal study in adults regarding dosage, wherein patients were randomized to receive taliglucerase alfa either 30 or 60 U/kg, every other week. However, in this study, the duration was 12 months instead of 9 months and the primary end point was improvement in hemoglobin rather than reduction in spleen volume. At the end of this study, clinically significant improvements were observed in hemoglobin concentration, platelet counts, spleen volume, and liver volume, as well as in GD biomarkers.

A novel mutation, p.N440del, localized to the TNAP crown domain was identified in the probands in this study. A number of TNAP missense mutations affecting amino acid residues localized in a flexible loop corresponding to the collagen-binding region have been identified in individuals diagnosed with HPP (Table 1). Moreover, Mornet et al. [13], showed that among 10 mutations localized in the crown domain and associated with HPP, at least six were located to this loop, including p.V423A, p.G426C, p.Y436H, p.S445P, p.R450C, and p.R450H. Interestingly, genetic alterations affecting amino acid residues

in the collagen-binding loop correspond to homozygous severe forms of HPP or heterozygous mild phenotypes (Table 1), indicating that this region plays an important, but presently unclear, role in

TNAP function. It is difficult to assign specific dysfunctions to each heterozygous mutation in these probands. Considering the functional and AZD9291 ic50 structural importance of the crown domain, and based on the pedigree, reduced ALP activity in vivo and in vitro, and predicted alterations in mutant TNAP structures, we hypothesize that the odonto-HPP phenotype is associated with the heterozygous deletion of residue N440, which may indirectly affect the enzyme activity, possibly from failure to reach a correct conformation, or via alterations in interactions with collagen. On the other hand, Selleckchem 3-Methyladenine based on TNAP protein expression and immunolocalization, analysis of internal contacts, and reports in the literature, we propose that the consequences of heterozygous N440 deletion are intensified by association with the heterozygous missense mutation, p.R152C. However, because the p.R152C mutation

was of maternal heritance and the mother was asymptomatic, we propose that when heterozygous and in the absence of other mutations, this alteration is not in itself sufficient Tenofovir research buy to significantly and deleteriously affect TNAP function. It remains unclear how particular ALPL mutations cause more severe clinical forms of HPP, in contrast to mild forms, including odonto-HPP. Dental tissues have been reported to be highly sensitive to dysregulation of phosphate and pyrophosphate metabolism [42], [43] and [44], perhaps indicating that less deleterious ALPL mutations may preferentially affect the dentition, but not the broader skeleton. We characterized a novel genetic alteration (c.1318_1320delAAC, p.N440del) in the ALPL gene resulting in odonto-HPP in the probands, monozygotic twins. Based on pedigree information, clinical symptoms, genetic analysis, and residual ALP activity, a genotype–phenotype association was established for p.N440del and odonto-HPP in this case. The heterozygous gene deletion was paternally inherited, and predicted to alter the loop harboring a collagen-binding site in the TNAP protein crown domain. In addition to this gene deletion, the probands feature an p.

, 2009). Therefore, the authors proposed that two of these genes (ednrb and sparc), which were found over-expressed in cavernosal tissue, were involved in priapism. mTOR inhibitor Ednrb gene directly activates the NO/cGMP pathway responsible for cavernosal

relaxation and consequently erection. Sparc gene is involved in many biological processes, such as vascular function, and could modulate cavernosal relaxation. Conversely, sod1 gene (superoxide dismutase 1) was suggested to be under-expressed after PnTx2-6 exposure, and this would have a negative effect in cavernosal cells, however this result was not confirmed by real time-PCR (Villanova et al., 2009). Functional experiments on cavernosal Selleck Gemcitabine strips using the pharmacological inhibitor ω-conotoxin GVIA (1 uM) and knockout mice (nNOS−/−, eNOS−/−) suggested that the relaxation promoted by PnTx2-6 depends on N-type Ca2+ channels in nitrergic nerve endings, which are the main source of NO release during erection (Nunes et al., 2010, 2012c). Furthermore, the same study showed that cavernosal relaxation improvement by PnTx2-6 does not depend on PDE-5 inhibition. In addition, it was shown that PnTx2-6 potentiates the CC relaxation by sildenafil, what suggests a different site to this toxin (Nunes et al., 2012b). This and other results suggest a local effect of PnTx2-6 (Fig. 2), although a central action for this toxin influencing the penile

erection could not be excluded. It is worth of note that the erection

with a purified toxin from P. nigriventer was firstly observed when PnTx2-6 was injected intracerebrally in mice (Dr. Carlos Diniz – personal for communication). In addition, mice injected intraperitoneally with this toxin showed an increase in c-fos activation in the paraventricular hypothalamus and in the nucleus of the stria terminalis, which are indirect markers for neuronal activity ( Troncone et al., 2011). These areas of the brain have been implicated in penile erection but are also related to stress. Although the crude venom has been shown to be able to cross the blood–brain barrier ( Le Sueur et al., 2003, 2004) it is not clear whether PnTx2-6 is able to do so. Thus, these data are not sufficient to support the involvement of the central nervous system in the pro-erectile action of PnTx2-6. Recent in vivo and in vitro studies showed that PnTx2-6 toxin improves cavernosal relaxation in different models of ED. In hypertensive patients, ED is a general complaint ( Javaroni and Neves, 2012), and these conditions seem to be connected ( Nunes et al., 2012a, Nunes and Webb, 2012). In DOCA– salt (deoxycorticosterone-acetate) rats, mineralocorticoide-induced hypertensive animals, which are well known models for hypertension and ED ( Chitaley et al., 2001), subcutaneous injection of PnTx2-6 toxin reversed severe ED.

Collectively, such findings have fostered the emergence of CSFs as a potential tool for the treatment of IBD ( Barahona-Garrido and Yamamoto-Furusho,

2008) and, in fact, recent controlled clinical trials have shown treatment with recombinant human GM-CSF to decrease disease severity and improve the quality of life of patients with active CD ( Goldstein et al., 2011 and Korzenik et al., 2005). It follows, therefore, that Roxadustat the retinoid-induced release of GM-CSF reported here, as distinct from LPS-induced responses, would provide potential benefit to the GI environment, particularly in pathological states such as IBD. A similar view could be taken regarding the observed changes in MCP-1. This key target, together with IL-10, is crucial for the GSK458 molecular weight regulation of immune responses against commensal bacteria by intestinal macrophages (Takada et al., 2010) and has been shown also to exert a beneficial effect on dextran sodium sulfate (DSS)-induced colitis in mice (Maharshak et al., 2010). Thus, as for GM-CSF, the retinoid-induced

release of MCP-1 seen in this study, both in the presence and absence of LPS, may similarly preclude a beneficial effect of this chemokine in steady-state gut homeostasis. In contrast, however, overexpression of VEGF-A has been shown to be associated with deterioration in disease status in mice with DSS-induced colitis, levels correlating with increased angiogenesis and leukocyte adhesion in the intestine (Scaldaferri et al., 2009), while increased levels of VEGF are usually observed in human subjects with IBD (Tsiolakidou et al., 2008). The release of VEGF might, therefore, be expected to convey potentially negative effects on intestinal www.selleck.co.jp/products/obeticholic-acid.html immunology. To counterbalance this argument, VEGF has also been observed to inhibit the apoptosis of intestinal epithelial cells – thus preventing bacterial translocation across ileal mucosa (Nakajima et al., 2007) – while levels of VEGF expression are reported as not being

associated with disease activity in patients with IBD (Alkim et al., 2012). Nevertheless, until more data become available relating to the effect of VEGF on maintenance of gut homeostasis, it is perhaps prudent that caution is exercised in assessing the overall effect of this cytokine target on the intestinal milieu. All retinoids tested were also found to have little or no adverse effect on the permeability of Caco-2 monolayers. This was also evident at all doses tested and is in apparent conflict with a relatively early in vitro study, which showed that the permeability of the Caco-2 monolayer, as measured by transepithelial electric resistance and [3H]-mannitol flux, was enhanced by ATRA. Given the known association between vitamin A deficiency and impairment in intestinal integrity, the authors considered this surprising and attributed increased permeability to an unknown mechanism(s) and not altered tight-junction protein expression ( Baltes et al., 2004).