ABSTRACT
Blueberries (Vaccinium corymbosum) undergo significant texture changes during development and ripening, notably a consistent decrease in firmness, which affects fruit quality, consumer preference, transportability, and shelf life. This study examined the composition and structural modifications of the cell wall in five commercially available blueberry varieties with differing firmness levels at harvest. Our approach integrated various biochemical techniques for a comprehensive analysis of cell wall components to elucidate firmness differences at the harvest stage. One of the conclusions was the relationship between a low degree of pectin methylesterification and the presence of increased egg-box structures, which correlated with increased firmness. The data suggest that low-abundance pectins in blueberry cell walls, such as rhamnogalacturonan-I participate in firmness modulation through their side branches or by linking to arabinogalactan proteins. Additionally, the xyloglucan structure can be one of the determinants of fruit firmness. Although, this work provides a broad insight into the relationship between cell wall composition and firmness in blueberry, a more detailed analysis, specifically focusing on pectin and hemicelluloses, would be of significant value.
ABSTRACT
Lichens are symbiotic organisms composed of at least one fungal and one algal species. They are found in different environments around the world, even in the poles and deserts. Some species can withstand extreme abiotic conditions, including radiation and the vacuum of space. Their chemistry is mainly due to the fungal metabolism and the production of several secondary metabolites with biological activity, which have been isolated due to an increasing interest from the pharmaceutical community. However, beyond the experimental data, little is known about their mechanisms of action and the potential pharmaceutical use of these kinds of molecules, especially the ones isolated from lesser-known species and/or lesser-studied countries. The main objective of this review is to analyze the bibliographical data of the biological activity of secondary metabolites from lichens, identifying the possible mechanisms of action and lichen species from Chile. We carried out a bibliographic revision of different scientific articles in order to collect all necessary information on the biological activity of the metabolites of these lichen species. For this, validated databases were used. We found the most recent reports where in vitro and in vivo studies have demonstrated the biological properties of these metabolites. The biological activity, namely anticancer, antioxidant, and anti-inflammatory activity, of 26 secondary metabolites are described, as well as their reported molecular mechanisms. The most notable metabolites found in this review were usnic acid, atranorin, protolichesterinic acid, and lobaric acid. Usnic acid was the most investigated metabolite, in addition to undergoing toxicological and pharmacological studies, where a hepatotoxicity effect was reported due to uncoupling oxidative phosphorylation. Additionally, no major studies have been made to validate the pharmacological application of these metabolites, and few advancements have been made in their artificial growth in bioreactors. Despite the described biological activities, there is little support to consider these metabolites in pharmaceutical formulations or to evaluate them in clinical trials. Nevertheless, it is important to carry out further studies regarding their possible human health effects. These lichen secondary metabolites present a promising research opportunity to find new pharmaceutical molecules due to their bioactive properties.
ABSTRACT
Medicinal plants have been used since prehistoric times and continue to treat several diseases as a fundamental part of the healing process. Inflammation is a condition characterized by redness, pain, and swelling. This process is a hard response by living tissue to any injury. Furthermore, inflammation is produced by various diseases such as rheumatic and immune-mediated conditions, cancer, cardiovascular diseases, obesity, and diabetes. Hence, anti-inflammatory-based treatments could emerge as a novel and exciting approach to treating these diseases. Medicinal plants and their secondary metabolites are known for their anti-inflammatory properties, and this review introduces various native Chilean plants whose anti-inflammatory effects have been evaluated in experimental studies. Fragaria chiloensis, Ugni molinae, Buddleja globosa, Aristotelia chilensis, Berberis microphylla, and Quillaja saponaria are some native species analyzed in this review. Since inflammation treatment is not a one-dimensional solution, this review seeks a multidimensional therapeutic approach to inflammation with plant extracts based on scientific and ancestral knowledge.
ABSTRACT
Chilling injury is a physiological disorder caused by cold storage in peaches and nectarines. The main symptom of chilling injury is mealiness/wooliness, described as a lack of juice in fruit flesh. In this work, we studied two nectarine varieties (Andes Nec-2 and Andes Nec-3) with contrasting susceptibility to mealiness after cold storage. A non-targeted metabolomic analysis was conducted by GC-MS to understand if changes in metabolite abundance are associated with nectarine mealiness induced by cold storage. Multivariate analyses indicated that in unripe nectarines, cold storage promoted a higher accumulation of amino acids in both varieties. Interestingly, for ripe nectarines, cold storage induced an accumulation of fewer amino acids in both varieties and showed an increased abundance of sugars and organic acids. A pathway reconstruction of primary metabolism revealed that in ripe nectarines, cold storage disrupted metabolite abundance in sugar metabolism and the TCA cycle, leading to a differential accumulation of amino acids, organic acids, and sugars in mealy and juicy nectarines.
ABSTRACT
Algae and microalgae are used as a source of different biomolecules, such as lipids and carbohydrates. Among carbohydrates, polysaccharides, such as ß-glucans, are important for their application as antioxidants, antisepsis, and immunomodulators. In the present work, the ß-glucans production potential of Microchloropsis salina was assessed using two different culture conditions: a high-density batch and a modeled high-density fed-batch. From the biochemical parameters determined from these two cultures conditions, it was possible to establish that the modeled high-density fed-batch culture improves the biomass growth. It was possible to obtain a biomass productivity equal to 8.00 × 10-2 ± 2.00 × 10-3 g/(L × day), while the batch condition reached 5.13 × 10-2 ± 4.00 × 10-4 g/(L × day). The same phenomenon was observed when analyzing the ß-glucans accumulation, reaching volumetric productivity equal to 5.96 × 10-3 ± 2.00 × 10-4 g of product/(L × day) against the 4.10 × 10-3 ± 2.00 × 10-4 g of product/(L × day) obtained in batch conditions. These data establish a baseline condition to optimize and significantly increase ß-glucan productivity, as well as biomass, adding a new and productive source of this polymer, and integrating its use in potential applications in the human and animal nutraceutical industry.
ABSTRACT
Tissue texture influences the grape berry consumers acceptance. We studied the biological differences between the inner and outer mesocarp tissues in hard and soft berries of table grapes cv NN107. Texture analysis revealed lower levels of firmness in the inner mesocarp as compared with the outer tissue. HPAEC-PAD analysis showed an increased abundance of cell wall monosaccharides in the inner mesocarp of harder berries at harvest. Immunohistochemical analysis displayed differences in homogalacturonan methylesterification and cell wall calcium between soft and hard berries. This last finding correlated with a differential abundance of calcium measured in the alcohol-insoluble residues (AIR) of the inner tissue of the hard berries. Analysis of abundance of polar metabolites suggested changes in cell wall carbon supply precursors, providing new clues in the identification of the biochemical factors that define the texture of the mesocarp of grape berries.
Subject(s)
Vitis , Calcium/metabolism , Cell Wall/chemistry , Fruit/metabolism , Metabolomics , Vitis/chemistryABSTRACT
Peumus boldus is an endemic tree species from Chile whose leaves have been the focus of study for decades given that their infusions are reported to relieve rheumatic symptoms, headache, dyspepsia, urinary tract inflammation, and symptoms of other illnesses. These health properties have been studied mainly using leaves and bark, then it is relevant to know more about these properties in different parts of the plant. Considering the importance of P. boldus fruits in the diet of some rural populations, we analyzed their properties to explore its impact on the Chilean population health. Liquid chromatography and mass spectrometry analysis confirmed the presence of alkaloids such as boldine, although aporphine N-methyl-laurotetanine was the most abundant. In addition, flavonoids catechin, chrysin and quercetin were also found in the extract. Cytotoxicity and anti-inflammatory activities of the fruit extract were invitro tested by using a murine macrophage cell model, observing that a diluted fraction of the extract was not cytotoxic, but showed anti-inflammatory activity, which is likely attributed to antioxidants activities. By means of quantum chemical calculations, we calculated the redox potential of the respective alkaloids and flavonoids found in the extract. Results suggest a synergistic effect between alkaloids and flavonoids, where boldine and N-methyl-laurotetanine showed similar antioxidant properties. Finally, we present a description of the oxidation mechanisms for both groups of molecules which will sustain P. boldus fruit biological properties, in order to give this kind of fruits scientific value focusing on human health.
Subject(s)
Peumus , Animals , Antioxidants/pharmacology , Fruit , Humans , Mice , Plant Extracts/pharmacology , Plant LeavesABSTRACT
The endoplasmic reticulum (ER) is the organelle where one third of the proteins of a cell are synthetized. Several of these proteins participate in the signaling and response of cells, tissues, or from the organism to the environment. To secure the proper synthesis and folding of these proteins, or the disposal of unfolded or misfolded proteins, the ER has different mechanisms that interact and regulate each other. These mechanisms are known as the ER quality control (ERQC), ER-associated degradation (ERAD) and the unfolded protein response (UPR), all three participants of the maintenance of ER protein homeostasis or proteostasis. Given the importance of the client proteins of these ER mechanisms in the plant response to the environment, it is expected that changes or alterations on their components have an impact on the plant response to environmental cues or stresses. In this mini review, we focus on the impact of the alteration of components of ERQC, ERAD and UPR in the plant response to abiotic stresses such as drought, heat, osmotic, salt and irradiation. Also, we summarize findings from recent publications looking for a connection between these processes and their possible client(s) proteins. From this, we observed that a clear connection has been established between the ERAD and UPR mechanisms, but evidence that connects ERQC components to these both processes or their possible client(s) proteins is still lacking. As a proposal, we suggest the use of proteomics approaches to uncover the identity of these proteins and their connection with ER proteostasis.
ABSTRACT
The firmness of blueberry is one of its most significant quality attributes. Modifications in the composition of the cell wall have been associated with changes in the fruit firmness. In this work, cell wall components and calcium concentration in two blueberry cultivars with contrasting firmness phenotypes were evaluated at harvest and 30 days cold storage (0 °C). High performance anion-exchange chromatography with pulse amperometric detector (HPAEC-PAD) analysis was performed using the "Emerald" (firmer) and "Jewel" (softer) blueberry cultivars, showing increased glucose in the firmer cultivar after cold storage. Moreover, the LM15 antibody, which recognizes xyloglucan domains, displayed an increased signal in the Emerald cultivar after 30 d cold storage. Additionally, the antibody 2F4, recognizing a homogalacturonan calcium-binding domain, showed a greater signal in the firmer Emerald blueberries, which correlates with a higher calcium concentration in the cell wall. These findings suggest that xyloglucan metabolism and a higher concentration of cell wall calcium influenced the firmness of the blueberry fruit. These results open new perspectives regarding the role of cell wall components as xyloglucans and calcium in blueberry firmness.
ABSTRACT
Psoriasis is a chronic, autoimmune, and inflammatory disease that affects 2% of the world's population. In recent years, it has been demonstrated that psoriasis confers a 25% increase in relative risk of cardiovascular disease, independent of factors such as hyperlipidemia, smoking, and obesity. The objective of this review was to analyze and describe the association between psoriasis and cardiovascular disease. In this review, we describe the epidemiological association of psoriasis and cardiovascular disease, pathophysiology, mechanisms, and its association with the well-known cardiovascular risk calculators. In addition, we describe diagnostic tools, such as imaging techniques and novel biomarkers, that are useful in the evaluation of atherosclerotic cardiovascular disease. Finally, we present different systemic therapies that are used in patients with psoriasis and their effect on atherosclerotic cardiovascular disease. This article provides an overview of the current literature on psoriasis and cardiovascular risk, which can be useful for primary care physicians in their daily clinical practice.
ABSTRACT
Frontal fibrosing alopecia (FFA) is a cicatricial alopecia characterized by hairline recession. Multiple autoimmune pathologies have been reported in patients with FFA. Despite the fact that FFA etiology remains unknown, there has been described an association with autoimmune disorders probably caused by an altered activity of cytotoxic CD8 T lymphocytes. Moreover, other autoimmune pathologies develop TH1 and TH17 response. Genetics could be responsible, in part, for the role of multiple simultaneous autoimmune disorders. Herein, we describe a case of a female patient with vitiligo, lichen sclerosus, and autoimmune hypothyroidism who developed a pruritic band-like recession of the frontal hairline. More research is needed in this area since autoimmune events in these patients may not be a mere coincidence.
ABSTRACT
Patients who are immunocompromised or have cognitive or physical disabilities are at a higher risk of being affected with infections such as crusted scabies. This is a rare skin hyperinfestation by Sarcoptes scabiei var. hominis. The main characteristic of this dermatosis is a thick crust due to the high concentration of mites; in addition, other manifestations such as papules, excoriations, and burrows may be absent. In severe cases, thick yellow-brown crusts and plaques with deep fissures are present. Diagnosis can be made by observing mites, ova, or feces from skin scrapings. Multiple therapies can be used in patients with this condition. Management with patient isolation is important to prevent institutional outbreaks. This disease can have high mortality, primarily due to sepsis. Awareness of this condition and its serious consequences is important to reduce its mortality and morbidity.
Subject(s)
Sarcoptes scabiei/growth & development , Scabies/diagnosis , Scabies/pathology , Skin/pathology , Skin/parasitology , Adult , Animals , Female , HumansABSTRACT
Plants must defend themselves against pathogens. The defense response requires greater protein synthesis, which generates endoplasmic reticulum (ER) stress, yet failure to attenuate this stress has detrimental effects. WRKY7/11/17 transcription factors (TFs) are negative regulators of immunity since mutants are more resistant to Pseudomonas syringae pv tomato (Pst) infection. Here, we reveal a connection between ER-stress and the molecular mechanisms underlying the wrky mutant phenotype. The bZIP28 TF upregulates ER-chaperone expression (BiP1/2, ERdj3B, and SDF2) upon exposure of Arabidopsis to a bacterial defense elicitor, flagellin 22 (Flg22). Also, the activation of ER-chaperones is more sustained in double and triple wrky mutants treated with Flg22, suggesting that WRKY7/11/17 TFs downregulate these genes. Moreover, wrky mutants accumulate more bZIP28 transcripts in response to Flg22, indicating that WRKY7/11/17 transcriptionally repress this TF. Using Arabidopsis protoplasts, we also demonstrate that WRKYs bind to the bZIP28 promoter via W-box elements. Additionally, triple wrky mutants are more resistant, whilst bzip28 mutants are more susceptible, to Pst infection. Finally, we postulate a model of PAMP-Triggered Immunity regulation, where Flg22 activates bZIP28-signaling inducing the expression of ER-stress genes, as well as WRKY7/11/17 expression, which in turn inhibits PTI by downregulating bZIP28, controlling physiological responses in the Arabidopsis-Pst interaction.
Subject(s)
Arabidopsis Proteins/metabolism , Arabidopsis/metabolism , Arabidopsis/microbiology , Arabidopsis/genetics , Arabidopsis Proteins/genetics , Basic-Leucine Zipper Transcription Factors/genetics , Basic-Leucine Zipper Transcription Factors/metabolism , Gene Expression Regulation, Plant , Plant Immunity/genetics , Plant Immunity/physiology , Plants, Genetically Modified/genetics , Plants, Genetically Modified/metabolism , Plants, Genetically Modified/microbiology , Pseudomonas syringae/pathogenicity , Transcription Factors/genetics , Transcription Factors/metabolism , Unfolded Protein Response/genetics , Unfolded Protein Response/physiologyABSTRACT
Firm berries are highly appreciated by table grape consumers. Cell wall composition is one of the main factors influencing the firmness of table grape berries. Nevertheless, the biological factors driving changes in berry firmness remain unclear. In the present work, we evaluated the firmness of berries of Vitis vinifera cv. Thompson Seedless. We selected two orchards displaying contrasting berry firmness and evaluated polar metabolites and cell wall composition. Our results suggest that berries from the soft orchard exhibited a higher accumulation of sugars at veraison whereas berries from the hard orchard accumulated the same sugars at harvest plus a higher amount of glucose monosaccharide at the cell wall. Thus, this study opens new insights about a connection between metabolic and cell wall changes with fruit firmness in a table grape variety, suggesting that it is possible to use metabolomic tools to identify metabolic biomarkers associated with table grape berry firmness.
Subject(s)
Cell Wall/chemistry , Vitis/chemistry , Fruit , MetabolomicsABSTRACT
Atherosclerotic cardiovascular disease (ASCVD) is the leading cause of death in patients with rheumatoid arthritis (RA). Chronic inflammation and traditional risk factors increase cardiovascular risk (CVR) in these patients. Several CVR calculators are used in general population and in RA patients to predict cardiovascular outcomes and tailor therapy but the precision of these calculators in RA patients has yet to be determined. The aim of this study is to determine which risk calculator correlates best with carotid ultrasound (US) findings, specifically carotid plaque (CP) and carotid intima-media thickness (CIMT) in RA patients without clinical manifestations. This was a cross-sectional observational study relating CVR scores in RA patients with the presence of carotid US findings. A total of 97 patients 40 to 75 years old who fulfilled the 2010 ACR/EULAR and/or the 1987 ACR classification criteria for RA were selected. Clinical assessment of cardiovascular risk was performed using seven calculators and carotid US measurement of intima-media thickness and plaque. The tests with the highest sensitivity for CIMT were the Framingham BMI, Framingham lipids, ACC/AHA 2013, and QRISK2. In CP, the highest sensitivity was in QRISK2, SCORE, and ACC/AHA 2013. RA patients should be comprehensively evaluated to detect cardiovascular risk. Carotid US may be routinely recommended to detect subclinical atherosclerosis in RA patients. A lower cutoff point in CVR scales may be necessary to identify patients with a low and intermediate CVR to detect subclinical atherosclerosis earlier and personalize therapy.
Subject(s)
Arthritis, Rheumatoid/complications , Carotid Artery Diseases/diagnostic imaging , Carotid Intima-Media Thickness , Adult , Aged , Body Mass Index , Cardiovascular Diseases/etiology , Carotid Artery Diseases/etiology , Cross-Sectional Studies , Diabetes Mellitus, Type 2 , Female , Humans , Lipids/blood , Male , Middle Aged , Risk FactorsABSTRACT
Low temperatures, salinity, and drought cause significant crop losses. These conditions involve osmotic stress, triggering transcriptional remodeling, and consequently, the restitution of cellular homeostasis and growth recovery. Protein transcription factors regulate target genes, thereby mediating plant responses to stress. bZIP17 is a transcription factor involved in cellular responses to salinity and the unfolded protein response. Because salinity can also produce osmotic stress, the role of bZIP17 in response to osmotic stress was assessed. Mannitol treatments induced the transcript accumulation and protein processing of bZIP17. Transcriptomic analyses showed that several genes associated with seed storage and germination showed lower expression in bzip17 mutants than in wild-type plants. Interestingly, bZIP17 transcript was more abundant in seeds, and germination analyses revealed that wild-type plants germinated later than bzip17 mutants in the presence of mannitol, but no effects were observed when the seeds were exposed to ABA. Finally, the transcript levels of bZIP17 target genes that control seed storage and germination were assessed in seeds exposed to mannitol treatments, which showed lower expression levels in bzip17 mutants compared to the wild-type seeds. These results suggest that bZIP17 plays a role in osmotic stress, acting as a negative regulator of germination through the regulation of genes involved in seed storage and germination.
Subject(s)
Arabidopsis Proteins/metabolism , Arabidopsis/metabolism , Basic-Leucine Zipper Transcription Factors/metabolism , Gene Expression Regulation, Plant/physiology , Germination/physiology , Osmotic Pressure/physiology , Seeds/metabolism , Arabidopsis/genetics , Arabidopsis Proteins/genetics , Basic-Leucine Zipper Transcription Factors/genetics , Seeds/geneticsABSTRACT
Patients with rheumatoid arthritis (RA) have a high risk for comorbid conditions which increase mortality, hospital admissions, costs of care and inability. To evaluate the prevalence of comorbidities in Mexican mestizo patients with RA and determine the associated risk factors. Cross-sectional study in which RA patients admitted to our outpatient clinic were consecutively enrolled. We collected data regarding demographics, disease characteristics and comorbidities at the time of the patient's visit to the clinic. We analyzed 225 patients. Their mean age was 55.7 ± 8.3 years; disease duration, 9.5 (3.8-15.5) years; female gender, 93.8%; Disease Activity Score using 28 joints-C-reactive protein, 3 (2-4); methotrexate use, 84.9%; use of any other conventional disease modifying anti-rheumatic drug, 65.7%; use of biological agents, 8%. The most frequently associated diseases were: hypertension, 29.8%; dyslipidemia, 27.1%; osteoporosis, 19.1%; diabetes, 12.4%; hypothyroidism, 6.2%; solid malignancies 4.4%. Risk factors were also evaluated, the most prevalent was overweight in 101 (44.9%) of our patients. A total of 71 (31.6%) had obesity. We also detected high blood pressure in 12.4%, hyperglycemia in 27.1% and hyperlipidemia in 49.8%. Due to the high frequency of comorbidities among RA patients, it is important to follow existing recommendations for their timely detection and management. Cardiovascular diseases must be evaluated with priority. The initial evaluation should include a thorough examination to prevent the deleterious effect of comorbidities in RA.
Subject(s)
Arthritis, Rheumatoid/ethnology , Indians, North American , Adult , Aged , Antirheumatic Agents/therapeutic use , Arthritis, Rheumatoid/diagnosis , Arthritis, Rheumatoid/drug therapy , Comorbidity , Cross-Sectional Studies , Female , Humans , Male , Mexico/epidemiology , Middle Aged , Prevalence , Prognosis , Prospective Studies , Risk Factors , Time FactorsABSTRACT
Variability of the 10-year cardiovascular (CV) risk predicted by the Framingham Risk Score (FRS) using lipids, FRS using body mass index (BMI), Reynolds Risk Score (RRS), QRISK2, Extended Risk Score-Rheumatoid Arthritis (ERS-RA), and algorithm developed by the American College of Cardiology and the American Heart Association in 2013 (ACC/AHA 2013) according to the European League Against Rheumatism (EULAR) 2015/2016 update of its evidence-based recommendations for cardiovascular risk management in patients with rheumatoid arthritis (RA) has not been evaluated in Mexican mestizo patients. CV risk was predicted using six different risk calculators in 116 patients, aged 40-75, who fulfilled the ACR/EULAR 2010 classification criteria. Results were multiplied by 1.5 according to the EULAR 2015/2016 update. Global comparison of the risk predicted by all scales was done using the Friedman test, considering a P value of ≤0.05 as statistically significant. Individual comparison between the algorithms was made using the Wilcoxon signed-rank test, and a P value of ≤0.003 was considered statistically significant. All calculators showed to be different in the Friedman test (p ≤ 0.001). Median values of predicted 10-year CV risk were 11.02% (6.18-17.55) for FRS BMI; 8.47% (4.6-13.16) for FRS lipids; 5.55% (2.5-11.85) for QRISK2; 5% (3.1-8.65) for ERS-RA; 3.6% (1.5-9.3) for ACC/AHA 2013; and 1.5% (1.5-4.5) for RRS. ERS-RA showed no difference when compared against QRISK2 (p = 0.269). CV risk calculators showed variability among them and cannot be used indistinctly in RA-patients.
Subject(s)
Arthritis, Rheumatoid/complications , Cardiovascular Diseases/etiology , Aged , Arthritis, Rheumatoid/epidemiology , Cardiovascular Diseases/epidemiology , Cross-Sectional Studies , Female , Humans , Male , Mexico/epidemiology , Middle Aged , Risk Assessment/methodsABSTRACT
Atherosclerotic cardiovascular disease (ASCVD) is the leading cause of death in rheumatoid arthritis (RA) patients. Guidelines of the American College of Cardiology and the American Heart Association (ACC/AHA) 2013 and the Adult Treatment Panel III (ATP-III) differ in their strategies to recommend initiation of statin therapy. The presence of carotid plaque (CP) by carotid ultrasound is an indication to begin statin therapy. We aimed to compare the recommendation to initiate statin therapy according to the ACC/AHA 2013 guidelines, ATP-III guidelines, and CP by carotid ultrasound. We then carried out an observational, cross-sectional study of 62 statin-naive Mexican mestizo RA patients, aged 40 to 75, who fulfilled the 1987 or 2010 ACR/European League Against Rheumatism (EULAR) classification criteria. CP was evaluated with B-mode ultrasound. Cohen's kappa (k) was used to assess agreement between ACC/AHA 2013 guidelines, ATP-III guidelines, and the presence of CP, considering a p < 0.05 as statistically significant. Agreement was classified as slight (0.01-0.20), fair (0.21-0.40), moderate (0.41-0.60), substantial (0.61-0.80), and an almost perfect agreement (0.81-1.00). Slight agreement (k = 0.096) was found when comparing statin recommendation between CP and ATP-III. Fair agreement (k = 0.242) was revealed between ACC/AHA 2013 and ATP-III. Comparison between ACC/AHA 2013 and CP showed moderate agreement (k = 0.438). ACC/AHA 2013 guidelines could be an adequate and cost-effective tool to evaluate the need of statin therapy in Mexican mestizo RA patients, with moderate agreement with the presence of CP by ultrasound.