The objective of the present study was to evaluate the clinical, radiological, and functional outcomes of a subscapularis transthoracic surgical approach and a posterolateral surgical approach with debridement, bone graft fusion, and internal fixation for the treatment of upper thoracic tuberculosis.There is currently debate over the best surgical approach for the treatment of upper thoracic tuberculosis. Traditionally, the subscapularis transthoracic approach has been preferred; however, the posterolateral approach has gained popularity in the past few years.A prospective, consecutive cohort of 43 upper thoracic tuberculosis patients with a mean age of 39 years (range: 20–52 years) was followed up for a minimum of 12 months (range: 12–60 months). Patients were randomly divided into 2 groups. Group A (n = 21) was treated by the subscapularis transthoracic approach and group B (n = 22) was treated by the posterolateral approach. All cases were evaluated for clinical, radiological, and functional outcomes. Intraoperative blood loss, operative duration, intraoperative and postoperative complications, hospital stay, the cure rate, fusion time, and the Frankel scale were used for clinical and functional evaluation, whereas the kyphosis angle was used for radiological evaluation.P > 0.05). The average operative duration for Group B was lower than that of Group A. There were no significant differences in intraoperative blood loss, intraoperative and postoperative complications, hospital stay, grafted bone fusion time, or cure rate between groups (P > 0.05). The Cobb's angle correction rate for group B (68.5%) was significantly better than that of group A (30.9%). The neurological score showed significant postoperative improvement in both groups, with no significant difference between the groups.Grafted bones were fused by 10 months in all cases. There was no statistically significant difference between groups before surgery in terms of gender, age, segmental tuberculosis, erythrocyte sedimentation rate (ESR), Frankel scale, or Cobb's angle (The subscapularis transthoracic approach and the posterolateral approach with debridement, bone graft fusion, and internal fixation are both sufficient and satisfactory for the surgical treatment of upper thoracic tuberculosis. However, the posterolateral approach is superior to the subscapularis transthoracic approach in terms of surgical trauma, operative time, and kyphosis correction. Although employed for decades, this approach resulted in a higher incidence of postoperative infection, increased operation trauma, and longer recovery time. In 1982, Standefer et al9 reported a method for the treatment of a cervical thoracic tumor, which involved the splitting of the sternum and removal of the inner part of the clavicle. This method, however, can result in shoulder girdle weakness, clavicular nonunion, sternal nonunion, and retrosternal soft tissue injury. In 1987, Turner and Webb10 first presented the subscapularis transthoracic approach, which became a widely adopted fixation method for the treatment of upper thoracic tuberculosis due to its improved safety, convenience, and reliability. Unfortunately, this approach has many disadvantages, including larger trauma, complications that limit shoulder function, and lower kyphosis correction ability. Thus, controversy remains regarding the best anterior approach and posterior instrumentation for lesion removal, reliable fixation, and reduced surgical trauma.11Spinal tuberculosis is prevalent in many developing countries. It is the most dangerous form of bone and joint tuberculosis due to serious ensuing complications, such as severe neurological deficits, kyphotic deformities, and paraplegia.12 In recent years, following the development of posterior pedicle screw segmental instrumentation and the improvement of operation skills, this approach has become more widely accepted for the treatment of upper thoracic tuberculosis.The limitations of these approaches have driven some surgeons to advocate for a posterolateral approach to reveal upper thoracic. In 1894, Menard presented a posterolateral approach involving the removal of a section of the ribs for the surgical treatment of upper thoracic tuberculosis. This approach has now been employed for decades, with reports indicating that it is relatively safe and has satisfactory clinical effectiveness.The objective of this study was to compare the clinical, radiological, and functional outcomes of the subscapularis transthoracic approach and the posterolateral approach with debridement, bone graft fusion, and internal fixation for the surgical treatment of upper thoracic tuberculosis. The 2 approaches were evaluated in terms of intraoperative blood loss, operation duration, bony fusion, intraoperative and postoperative complications, hospital stay duration, cure rate, Frankel scale, and kyphosis angle.Forty-three consecutive patients with upper thoracic tuberculosis were evaluated in this prospective study. These patients were randomly divided into 2 groups according to their sequence of admission using a table of random numbers, with 21 patients in group A and 22 in group B. Even group (group A) was treated using the subscapularis transthoracic approach and odd group (group B) was treated using the posterolateral approach. The indications for surgery included patients with upper thoracic tuberculosis (T1 ∼ T4) for whom anti-tuberculosis drugs were ineffective and who may have also suffered from greater sequestrum formation, sinus formation, larger abscesses, serious vertebral collapse, progressive kyphosis, and neurological deficits. Surgical contraindications were patients with liver, heart, and kidney dysfunction; highly infectious tuberculosis; or serious underlying lung disease. The exclusion criteria included patients younger than 18 years with a history of upper thoracic surgery and incomplete follow-up information, and patients with a distant stream abscess that would require anterior surgery or multiple incisions.All patients in the study provided their written informed consent. This study was approved by the institutional review board and the ethics committee of the 175th Hospital of PLA.Standard laboratory tests were performed for all patients and included erythrocyte sedimentation rate (ESR), blood chemistry profiling, white and red blood cell count, and a Mantoux tuberculin skin test. An electrocardiogram, a radiograph of the chest, magnetic resonance imaging (MRI), and computerized tomography (CT) were also performed for all patients. Narrowing of the intervertebral space and different degrees of vertebral collapse were observed during X-ray examination for every patient. For every case, different degrees of sequestration were observed during CT examination, and varying degrees of vertebral body destruction were observed during MRI examination. Thirty-seven cases had varying degrees of spinal cord compression, and 35 cases exhibited paravertebral abscess. No patients had sinus formation.After the preliminary diagnosis of upper thoracic tuberculosis, all patients received standard anti-tuberculosis chemotherapy, including oral administration of pyrazinamide (0.5 g per day), isoniazid (0.3 g per day), rifampin (0.45 g per day), and ethambutol (0.75 g per day) for 2 to 4 weeks. ESR was tested on alternate weeks, and surgery was performed when the changes in ESR did not exceed 10 mm/h and did not display an increasing trend.For all patients in group A, subscapularis transthoracic surgery was performed under general anesthesia. First, patients were placed in the lateral position, and a midline incision was made from the C7 or T1 spinous process to the junction of the anterior axillary line and 4,5 intercostal via the medial of spealbone. Then, the muscle and fascia were incised, and the loose connective tissue was separated between the scapula and the chest wall. If necessary, surgeons exposed the lesions on the vertebra by resecting the ribs  and by using curettes to debride the affected vertebral body, infected tissue, pus, granulation tissue, sequestrum, and disc necrotic tissue. Third, an intervertebral bone autograft from the iliac crest and Ventrofix fixation system  was used. Streptomycin (2 g) was sprayed onto the operation site, and a local drainage tube was inserted before the incision was closed.In group B, all surgical operations were performed with the patients placed in a prone position under controlled general anesthesia with endotracheal intubation. All 22 patients were treated with a posterolateral surgical approach with debridement, bone graft fusion, and internal fixation. Through a midline incision, the posterior spinal construction was exposed, including the spinous processes, lamina, facet joints, and transverse processes. With C-arm fluoroscopy assistance, pedicle screws or hooks were placed at least 2 levels superior and inferior to the level of decompression. The spine was stabilized using a temporary fixed rod to avoid spinal cord injury caused by debridement and decompression. To drain prevertebral abscesses and to expose diseased vertebral bodies, surgeons performed a costotransversectomy and removed the articular process, pedicle, and transverse process on the more severely affected side of the lesion segment. Spinal tuberculosis was revealed by the separation of the posterior longitudinal ligament and dural sac. The posterior longitudinal ligament was then incised, and spatulas of various sizes and angles were used from a posterolateral approach to remove lesions, including sequestra, abscesses, and granulation tissue. An appropriately sized iliac bone was removed from the posterior superior iliac spine and was implanted in the interbody. If necessary, the intercostal nerve was cut off. Streptomycin was applied as for group A. Eight patients with obvious kyphosis were corrected by applying appropriate pressure. Drainage and incisional sutures were performed postoperatively.Blood pressure, respiration, pulse, amount of drainage, and sense and motor responses of the lower extremities were monitored after the operation. The drainage tube was removed when the amount of drainage was less than 10 mL per day. Mannitol was used for the treatment of edema for 3 to 5 days, while neurotrophic therapy was performed for patients with neurological dysfunction. All patients received antituberculosis chemotherapy with the 4 drugs mentioned above for 12 to 18 months. Postoperative and preoperative anti-tuberculosis treatment programs were the same. All patients were immobilized in a rigid external orthosis for 3 to 4 months after surgery. Follow-up examinations were performed for all patients at 1, 3, 6, 9, and 12 months after surgery, and subsequent follow-ups were conducted at 12-month intervals. At each follow-up evaluation, liver and kidney function, routine blood test, C-reactive protein, and ESR were monitored carefully to assess the presence of active disease. Routine lateral and anteroposterior X-ray, CT, and MRI imaging was performed to evaluate the extent of decompression, fusion status, functional nerve recovery, and Cobb's angle of kyphosis.13 The criteria for clinical cure of spinal tuberculosis included good general condition; absence of fever; normal appetite; absence of local pain; X-ray imaging of vertebral healing with a clear outline of the lesion; uniform MRI signal among the vertebral lesion, the surrounding tissue, and the normal vertebrae; and consistent normal ESR results. Patients were considered to have recurrence of spinal tuberculosis if the spinal tuberculosis was cured after surgery, but the primary lesion and corresponding symptoms reappeared after 1 year.For assessment of the correction rate, the kyphosis correction rate was defined as: (preoperative Cobb's angle – postoperative Cobb's angle)/preoperative Cobb's angle × 100%. Bone grafting fusion was assessed using the radiological criteria of Bridwell et al.t test were conducted for comparisons between groups A and B, and a P value <0.05 was considered statistically significant.All statistical analyses were performed using SPSS 21.0 software. Classified variables and continuous variables were expressed by frequency and means ± standard deviation, respectively. A Wilcoxon rank sum test and P > 0.05) with respect to their gender, age, segmental tuberculosis, ESR, Frankel scale, and Cobb's angle (Table P > 0.05), with no cases lost during the follow-up period. The follow-up time was no statistical significance between the 2 groups .From January 2008 to December 2013, we treated a total of 43 patients with upper thoracic tuberculosis by either a subscapularis transthoracic approach or a posterolateral approach. Patients in the 2 study groups were comparable (P < 0.05), while the differences in blood loss, length of hospital stay, grafted bone fusion time, and cure rate were not statistically significant (Table P = 0.306). For 3 cases that demonstrated recurrence, antituberculosis treatment was administered and surgical treatment with a posterolateral approach was performed after the ESR was stable.The surgical time was significantly shorter in group B than in group A  after surgery (P < 0.05). Group B exhibited a greater kyphosis correction , as evaluated by the Frankel scale . However, the difference between the 2 groups was not statistically significant .In group A, 2 cases with postoperative shoulder abduction movement disorder (abduction angles: 0 ∼ 90° and 0 ∼ 110°) were treated by rehabilitation therapy, and movement was recovered at the 6-month follow-up examination. There were no other complications, such as leakage of cerebrospinal fluid or nerve injury aggravation. The complication rate for group A was 9.5%. In group B, 2 patients exhibited leakage of cerebrospinal fluid; thus, the usage of drainage pipes was prolonged. There were no other complications, such as wound infection, shoulder movement disorder, or nerve injury aggravation. The complication rate for group B was 9.1%. The incidence of complications between the 2 groups was not statistically different . The kyphosis correction decreased by 6.8° in the anterior group and 6.1° in the posterior group when measured at the last follow-up examination (P < 0.01).There has been considerable debate over the best surgical approach for the treatment of upper thoracic tuberculosis. Surgical treatment of this disease must combine expertise in cervical lordosis and thoracic kyphosis with knowledge of the complex anatomy and important blood vessels, nerves, lymphatic vessels, and other structures within this anatomical region. There have been several anterior approaches reported in the literature, including a presternal approach10 However, this approach also has significant disadvantages, including insufficient kyphosis correction, larger surgical trauma, and increased postoperative complications. In this study, we believe that the subscapularis transthoracic approach need to cut the musculi pectoralis major, latissimus dorsi, trapezius, serratus anterior muscle, rhomboid muscle, and vertical spine muscle. This approach also resulted in a longer surgical time and larger operation wound than the posterolateral approach. A higher incidence of postoperative shoulder movement disorder occurred due to the separation of loose connective tissue between the scapula and the chest wall (9.5% incidence in this study). Moreover, the kyphosis correction was limited because the spinous process, lamina, and other rear structures could not be removed via the anterior approach. In our study, kyphosis correction was greater in in group B (68.5%) than in group A (30.9%), which agrees with the conclusion of Pinglin et al.18 Therefore, posterior surgery is more appropriate for patients with obvious kyphosis.A subscapularis transthoracic approach for the treatment of thoracic tuberculosis has many advantages, such as clear vision, avoidance of major blood vessels and nerves, and safe, adequate spinal decompression.12 The advantages of posterolateral approach include a single incision for the debridement, spinal decompression, bone graft and internal fixation; less operation trauma; low incidence of postoperative shoulder dysfunction; more reliable pedicle screw fixation; more effective kyphosis correction; and better recovery of physiological spine curvature. However, this approach also has some disadvantages, such as a longer fixed segment and the destruction of the structure of the rear normal spine. In this study, we found no significant difference in the recovery of neurological function between the 2 groups. We also found that the posterolateral approach can also be performed safely and effectively for spinal decompression.Menard first employed the posterolateral approach for the treatment of upper thoracic tuberculosis; subsequently, many confirmed the safety and efficacy of this approach.20 In our study, of the 22 patients with upper thoracic tuberculosis who were treated by the posterolateral approach, none exhibited failed internal fixation or failed fusion. Although the structure of the rear normal spine was destroyed via the posterolateral approach, the strength and long length of the segment pedicle fixation prevented any failure of the internal fixation or fusion, as evaluated during follow-up examinations. We believe that the posterolateral approach with resection of unilateral lamina, facet, transverse, and a small part of the ribs achieves effective postoperative fusion and does not cause upper thoracic instability. Possible reasons are the smaller mobility of the upper thoracic area relative to the cervical vertebra, lumbar vertebra and stronger capability of pedicle screw with good support, and anti-thoracic flexion, torsion. Intraoperative interbody fusion may also reestablish spinal stability.Some experts have argued that the posterolateral approach combined with lesion debridement is unsafe because it may destroy the healthy posterior spinal column, leading to spinal instability.In brief, the posterolateral approach is a feasible and effective method for the treatment of upper thoracic tuberculosis. In this study, it was superior to the subscapularis transthoracic approach. The posterolateral approach allowed for smaller surgical trauma and shorter operating time, and patients treated with this approach achieved remarkable kyphosis correction in their final follow-up evaluation. Due to the short follow-up times and low sample numbers in this study, there is a need for additional multicenter studies and studies with larger sample sizes to verify the clinical efficacy of this approach.Both surgical approaches improved patients’ Frankel scores and radiological results; however, there was no significant difference between the groups. This study provides evidence for the efficacy of both the subscapularis transthoracic approach and the posterolateral approach with debridement, bone graft fusion, and internal fixation for the surgical treatment of upper thoracic tuberculosis. The posterolateral approach offers a shorter operative time and better kyphosis correction; therefore, it may be a better surgical treatment for upper thoracic spinal tuberculosis.


Carotenoids are natural pigments that play pivotal roles in many physiological functions. The characteristics of carotenoids, their effects on health, and the cosmetic benefits of their usage have been under investigation for a long time; however, most reviews on this subject focus on carotenoids obtained from several microalgae, vegetables, fruits, and higher plants. Recently, microalgae have received much attention due to their abilities in producing novel bioactive metabolites, including a wide range of different carotenoids that can provide for health and cosmetic benefits. The main objectives of this review are to provide an updated view of recent work on the health and cosmetic benefits associated with carotenoid use, as well as to provide a list of microalgae that produce different types of carotenoids. This review could provide new insights to researchers on the potential role of carotenoids in improving human health. Due to changes in life style and the rising health consciousness of the average population, the demand for nutrient-rich supplements with health benefits (such as immunity boosters and supplements rich in proteins and vitamins) has risen. Carotenoids have various medicinal properties and they are widely used as preventives against diseases such as cancer, diabetes, and cataract [Carotenoids have a wide range of applications in the healthcare and nutraceuticals industry. The growing importance of carotenoid use in improving food quality has also led to an increase in the demand for carotenoids in the global market. The global carotenoid market was estimated to be ~1.24 billion USD in 2016, and is projected to increase to ~1.53 billion USD by 2021, at a compound annual growth rate (CAGR) of 3.78% from 2016 to 2021  in their structures. Xanthophylls or oxycarotenoids, which belong to the second group, are oxygen-containing molecules. Lutein and zeaxanthin are two xanthophylls with –OH groups in their structures, whereas canthaxanthin and echinenone contain =O groups. Astaxanthin has both –OH and =O groups in its structure. Furthermore, some carotenoids such as violaxanthin and diadinoxanthin contain epoxy groups, and others such as dinoxanthin and fucoxanthin have acetyl groups in their structures. The two carotenoids with acetyl groups also contain the C=C=C  group in their structures, which is unique to natural products . In addiIn plants, carotenoids play a significant role in the photosynthetic process by forming pigment–protein complexes in the thylakoid membrane; in cyanobacteria, however, they may also be found in the plasma membrane, and function in protecting the cells during exposure to high light intensities ,53. In aOur current understanding of the carotenoid metabolic pathway and its regulation in microalgae is unclear, and is mainly inferred from our knowledge of the process in higher plants. In chloroplasts, several nucleus-encoded membrane proteins are involved in the synthesis of carotenoids. These proteins are produced in the cytoplasm as precursor polypeptides with amino-terminal extensions that target them to chloroplasts. There are many publications that comprehensively review the carotenoid biosynthetic pathway ,62,63,64d-erythriol 4-phosphate (MEP) pathway [Carotenoids are synthesized from a C5 building block, a common precursor to all isoprenoids, which is the isopentenyl pyrophosphate (IPP) molecule, via the plastidial 2-C-methyl- pathway . The fir pathway . In anotIn most microalgae, the basic carotenoid synthesis pathways are highly conserved, although some species are able to accumulate unusual carotenoids via specific biosynthetic routes. In higher plants, most of the genes encoding enzymes for the carotenoid biosynthetic pathway have been isolated and characterized by functional complementation experiments . In micrAngiogenesis is the process of formation of new blood vessels from pre-existing capillaries and involves a sequence of events that are fundamental to many physiological and pathological processes . It occuStudies involving in vivo and in vitro experiments on male C57BL/6 mice and B16F-10 cells have been used to evaluate the anti-angiogenic effects of β-carotene by Guruvayoorappan and Kuttan . Their sIn another study, Sugawara et al.  reportedA study in 2005 by Hussein et al.  reportedHaematococcus pluvialis on human blood rheology were carried out on 20 adult men. After 10 days of astaxanthin (6 mg/day) administration, the whole blood transit time of the experimental group decreased from 52.8 ± 4.9 s to 47.6 ± 4.2 s, which is considerably lower than that of the control group (54.2 ± 6.7 s) [In another study, female BALB/c mice treated with 800 mg/kg astaxanthin for eight weeks exhibited higher heart mitochondrial membrane potentials and contractility indices than mice in a placebo group . An ex v± 6.7 s) . In addi± 6.7 s)  showed t± 6.7 s) . FucoxanNumerous in vitro and in vivo studies have demonstrated the anti-cancer activities of carotenoids. The results of these studies indicate that carotenoids may prevent different types of cancers in humans, including bladder, breast, hepatic, intestinal, leukemic, lung, oral, and prostate cancer. The anti-cancer activity of carotenoids involves a variety of mechanisms, including induction of cell apoptosis and suppression of cell proliferation. In particular, one in vivo study showed that β-carotene, astaxanthin, canthaxanthin, and zeaxanthin help in reducing the sizes and numbers of liver neoplasias . AnotherMany studies indicate that β-carotene shows great potential as an anti-tumor agent. In a study in China, administration of a combination of β-carotene, vitamin E, and selenium to humans was observed to decrease the incidence of mortality due to cancer . Many otThe anti-proliferative and cancer-preventive activities of fucoxanthin and fucoxanthinol are dependent on different molecules and pathways involved in the processes of cell cycle arrest, apoptosis, and metastasis . FurtherThe effect of different carotenoids such as β-cryptoxanthin, canthaxanthin, fucoxanthin, neoxanthin, phytoene, and zeaxanthin on the viabilities of three prostate cancer cell lines, namely, PC-3, DU 145, and LNCaP, has been investigated . Among tCanthaxanthin, which is another type of carotenoid with significant anti-cancer activity, has been reported to significantly inhibit the growth of JB/MS, B16F10 (a melanoma cell line), and PYB6 (a fibrosarcoma tumor cell line) cells at a concentration of 100 mM . TreatmeSeveral studies have reported that astaxanthin has significant anti-cancer effects on certain cancer types such as prostatic hyperplasias and prostatic cancers. Astaxanthin mainly inhibits the enzyme 5-α-reductase, which is involved in abnormal prostate growth ,104. TheAn investigation by Lyons and O’Brien  on the dThe anti-cancer properties of the carotenoid cryptoxanthin have not been investigated extensively. However, recent in vitro, in vivo, and human-intervention studies report that β-cryptoxanthin can differentially regulate the expression of P73 variants. In addition, these studies have found that this carotenoid has the ability to inhibit the proliferation of colon cancer cells and in conjunction with oxaliplatin, can induce apoptosis in cancer cells by negatively regulating ΔNP73 .Recent work on carotenoids suggests that these molecules may be more effective in treating and controlling diabetes than antioxidants. Studies have shown that levels of dietary carotenoids and concentrations of β-carotene in blood are inversely associated with fasting blood glucose levels and insulin resistance, respectively . NumerouMost studies on carotenoids and diabetes report the importance of carotenoids in dietary intake for the prevention and treatment of T2DM . A recenAstaxanthin, which is one of the best studied carotenoids, shows great potential in preventing and treating diabetes. Astaxanthin has higher antioxidant activity than other carotenoids such as lutein, β-carotene, and zeaxanthin , and canAnother important carotenoid, β-carotene, has been investigated in detail regarding its usefulness in the treatment of diabetes. Hozumi et al.  reportedKKAy mice. Similarly, it has also been demonstrated that in KKAy mice, fucoxanthin can reduce hyperglycemia, although this carotenoid has no such effect on lean C57BL/6J mice [Another carotenoid, fucoxanthin, shows good potential as an anti-diabetic agent. A study on fucoxanthin reports that treatment with this carotenoid can restore blood glucose and insulin levels to normal in obese mice. The study reports that fucoxanthin upregulated the genetic expression and mRNA levels of the glucose transporter 4 (GLUT4) protein in skeletal muscle cells . Nishika/6J mice . Other s/6J mice . In addi/6J mice . From th2 (PGE2), and proinflammatory cytokines, as well as their inhibitory effects on enzymes such as inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (cox-2) [The first response of the immune system to infection or irritation is inflammation, which is also referred to as the innate cascade. However, some inflammatory reactions can have adverse effects on host cells or tissues; for example, chronic inflammation can cause many conditions such as arthritis, hepatitis, gastritis, periodontal disease, colitis, atherosclerosis, pneumonia, and neuro-inflammatory diseases . Therefo (cox-2) ,140.2, TNF-α, and interleukin-1β (IL-1β), as well as by blocking the activity of NOS enzymes in RAW 264.7 cells. The results of this study agree with those of a previously conducted study, which showed that astaxanthin inhibited NO production, as well as the expression of iNOS and COX-2 in LPS-stimulated BV2 microglial cells [Recently, astaxanthin has garnered much attention due to its potential as an anti-inflammatory agent. Both, in vitro and in vivo studies have been carried out in rat models to investigate the effects of astaxanthin on lipopolysaccharide (LPS)-induced inflammatory reactions ,142. Theal cells .2, as well as the expression of pro-inflammatory genes by suppressing the function of NF-κB. Furthermore, astaxanthin also suppressed the activity of the iNOS promoter by inhibiting IKK (IκB kinase) activity. A similar study found that LPS-stimulated mouse neutrophils treated with astaxanthin produce significantly lower levels of the proinflammatory cytokines TNF-α and IL-6. Macedo et al. [2O2. Cytokine levels in cells pre-incubated with 10 mM astaxanthin before H2O2 stimulation were significantly lower  of the levels seen in control cells (cells not pre-treated with astaxanthin). Furthermore, cells pre-incubated with astaxanthin also showed restoration of SHP-1 (a protein tyrosine phosphate) expression levels and reduced levels of NF-κB expression [Helicobacter pylori-infected mice fed with astaxanthin extracted from the microalga H. pluvialis, showed reduced levels of gastric inflammation. Mice fed with 200 mg/kg of algal extract for 10 days, showed significantly lower levels of inflammation and mucosa-bacterial loads in their stomachs than untreated mice. These results indicate a change in the T-lymphocyte response in mice; the response changes from a predominantly Th1-response to a mixed Th1/Th2-response. This shift was found to occur because of a block in IFN-γ release that boosts IL-4 release in splenocytes in the infected mice pre-treated with astaxanthin. A clinical study to investigate the anti-oxidative and anti-inflammatory effects of astaxanthin was also undertaken on a cohort of healthy young women [Lee et al.  also repo et al.  also reppression . Bennedspression  reportedng women . In thisng women . OverallObesity is a condition where excessive accumulation and storage of fat in the body occurs, leading to inordinate increases in body weight . ObesityWang et al.  reportedMaeda et al.  used a mROS and reactive nitrogen species (RNS) are generated during aerobic metabolism processes that occur in the cell; these include processes such as signal transduction, gene expression, and activation of cell signaling cascades . ROS can1O2) and peroxyl radicals, but there is little information regarding their roles in cellular defenses against RNS. Raposo et al. [−. Several studies have reported that dietary intake of carotenoids can protect humans and animals from oxidative damage to lipophilic parts of cells; this is because carotenoids can limit lipid peroxidation events by scavenging the ROS formed during photo-oxidative processes [Spirulina platensis, H. pluvialis, and Dunaliella salina might be of great value in the production of various types of such carotenoids  is one of the leading causes of visual impairment. In the United States of America, approximately 2.07 million Americans were affected with advanced ARMD in 2010, and this number is expected to increase to 5.44 million in 2050. ARMD is a major cause of irreversible blindness among elderly people (>65 years of age) in western countries, and affects ~20% of the total population . The macAn investigation using unilamellar liposomes as a model membrane showed that the filtering effects of lutein and zeaxanthin are higher than those of lycopene and β-carotene. Because of this, lutein and zeaxanthin are thought to be essential pigments in the lens and retina of the eye, and maintaining their levels could be critical in protecting vision in older people . AlthougNeuroprotection strategies are defined as the mechanisms and strategies used to protect neuronal cells against injury, apoptosis, dysfunction and/or degeneration in the central nervous system (CNS) by limiting neuronal dysfunction or death after CNS injury . As mostOsteoclasts are highly specialized bone cells that break down bone tissues. One of the most recent uses of fucoxanthin has been in the treatment of osteoclast diseases. Das et al.  reportedFucoxanthin is a well-known weight-loss agent that encourages the ‘burning of fats’ by enhancing thermogenin expression. As fucoxanthin has numerous health benefits, and the molecule is found in high concentrations in microalgae, industrial production of fucoxanthin from microalgae is on the rise . In 2009Microalgae are rich sources of carotenoids, with great industrial potential and accessibility, and thus are likely to have a wide range of applications in the healthcare and cosmetic industries. Most studies on carotenoids have been focused mainly on the preventive and protective effects of these molecules in various chronic diseases such as diabetes mellitus, metabolic syndrome, cancer, and cardiovascular diseases. Recent studies, however, report that carotenoids might also play a significant role in the treatment of various other diseases. Although the mechanisms of antioxidant activity for some carotenoids have been well studied, most of the other effects of carotenoids, such as their pro-vitamin A activity, metabolic activity, effects on the immune and endocrine systems, as well as their effects on cell cycle regulation, apoptosis, and cell differentiation have not yet been studied in detail. Although there are a number of ongoing studies investigating the use of carotenoids to enhance healthcare and beauty, most of these studies have been carried out in animal models, with very few human clinical trials. Future areas of research will need to focus on human clinical trials. In addition, these studies must collect detailed data on subject selection, end point measurements and levels of carotenoids being tested. It is hoped that such studies, will help researchers understand the roles and potential uses of carotenoids in developing new strategies for the prevention, treatment, and management of diseases.


Currently, the management of urban waste streams in developing countries is not optimized yet, and in many cases these wastes are disposed untreated in open dumps. This fact causes serious environmental and health problems due to the presence of contaminants and pathogens. Frequently, the use of specific low-cost strategies reduces the total amount of wastes. These strategies are mainly associated to the identification, separate collection and composting of specific organic waste streams, such as vegetable and fruit refuses from food markets and urban gardening activities. Concretely, in the Chimborazo Region (Ecuador), more than 80% of municipal solid waste is dumped into environment due to the lack of an efficient waste management strategy. Therefore, the aim of this study was to develop a demonstration project at field scale in this region to evaluate the feasibility of implanting the composting technology not only for the management of the organic waste fluxes from food market and gardening activities to be scaled-up in other developing regions, but also to obtain an end-product with a commercial value as organic fertilizer. Three co-composting mixtures were prepared using market wastes mixed with pruning of trees and ornamental palms as bulking agents. Two piles were created using different proportions of market waste and prunings of trees and ornamental palms: pile 1 (50:33:17) with a C/N ratio 25; pile 2: (60:30:10) with C/N ratio 24 and pile 3 (75:0:25) with C/N ratio 33), prepared with market waste and prunings of ornamental palm. Throughout the process, the temperature of the mixtures was monitored and organic matter evolution was determined using thermogravimetric and chemical techniques. Additionally, physico-chemical, chemical and agronomic parameters were determined to evaluate compost quality. The results obtained indicated that all the piles showed a suitable development of the composting process, with a significant organic matter decomposition, reached in a shorter period of time in pile 3. At the end of the process, all the composts showed absence of phytotoxicity and suitable agronomic properties for their use as organic fertilizers. This reflects the viability of the proposed alternative to be scaled-up in developing areas, not only to manage and recycle urban waste fluxes, but also to obtain organic fertilizers, including added value in economic terms related to nutrient contents. The increasing generation of food waste and its management constitute a major concern. In the EU, around 100 million tonnes of food, mainly coming from homes but also from food markets, are wasted annually. This amount is expected to rise to over 120 million tonnes by 2020 if the situation does not change . This fa2O5-K2O nutrient values in food market wastes from the studied area . In addition, a study in six markets in Morelia (Mexico) concluded that biodegradable compounds in food market waste were around 93% [In the biggest market of the city of Riobamba , as in other regions of Ecuador, different products that include vegetables, cereals, tubers, fruits, meat, dairy and foods in general are sold, mainly at whole scale. In this market, 6 tonnes of waste are daily generated, 95% 5.7 tonnes) being organic waste, which are currently disposed without any treatment into the open dump of the city (Data from the Departamento de Desechos Sólidos en Riobamba). The wastes from food market, principally constituted by vegetable waste, usually contain high levels of organic matter (OM), moisture, and nutrients  tonnes b reportedound 93%  reportedound 93% . This reound 93% . In manyound 93% , 14. ThiTherefore, the management of these waste fluxes must be quick and efficient due to their high degradability, which makes unsuitable their disposal in municipal landfills. The disposal in landfills or dumpsites of these organic wastes, especially vegetable market waste, together with municipal solid wastes, generates different undesirable effects and negative impacts in the environment  that affect the health of the population of the area of influence . HoweverTherefore, the main aim of this study were twofold: i) to evaluate and develop different co-composting strategies based on the use of food market wastes and pruning waste from urban gardening activities ; ii) to obtain high quality composts under the conditions of the studied region, including an economical assessment of the NPK value content in these organic fertilizers.Acacia L., Ficus L. and Populus L.) of the avenues and parks of the city. PP came from ornamental palms (Phoenix canariensis). MW, TP and PP were homogenised and crushed using a crusher to obtain a particle size between 1 and 4 cm  were collected in the San Pedro de Riobamba market (known as Mercado Mayorista), which is located in the south of the city of Riobamba . The wastes were collected directly from vendors, with the authorisation of the Gobierno Autónomo Descentralizado Municipal de Riobamba, and were fresh vegetables remains: onion, artichoke, broccoli, spinach, cabbage, lettuce, garlic, basil, carrot, beet, cabbage, cauliflower, celery, radish, pepper, bean, corn, bean, pea, potato, and fruits such as apple, strawberry, blackberry, orange, banana, tomato, pineapple, mango, peach, melon, watermelon, lemon, avocado, grape, papaya, kiwi, plum, fig, lime. The improper materials, such as plastic, glass and paper were manually separated. Pruning wastes, tree prunings (TP) and palm prunings (PP) were collected separately in order to use them as bulking agents in the co-composting process. TP contained leaves and young branches of ornamental trees and shrubs dry :The study was carried out at the open composting facility of the Environmental Theme Park Ricpamba (Riobamba-Ecuador). The mixtures (about 1000 kg each) were managed as trapezoidal windrows (about 1.8 x 3 x 1.5 m (length x width x height) and were mechanically turned when the temperature dropped to less than 40°C , in order to homogenise and aerate the mixtures. The bio-oxidative phase of composting was considered finished when during 10 consecutive days after a whirl the difference between the pile temperature and the ambient temperature was ≤ 10°C. Then, composts were left to mature in static conditions over a period of one month, approximately. Throughout the composting process, temperature was monitored daily and the humidity of each pile was kept at optimal level for microbial metabolism (40–60%), by adding the necessary water. EXI was calculated as the summation of the daily value obtained by subtracting the ambient temperature from the temperature value in the composting pile during the bio-oxidative phase of composting, and expressed as cumulated °C. Composite samples were collected throughout the composting process, by taking, mixing and homogenizing ten subsamples from the whole profile of each mixture (from the top to bottom). All the samples were air-dried and ground to 0.5 mm prior to analysis.3/HClO4 digestion, phosphorus (P) was assessed colorimetrically as molybdovanadate phosphoric acid, and potassium (K) was determined by flame photometry . The germination Index (GI) was calculated using seeds of radish (Raphanus sativus L.) [2-triethanolamine according to the method described by [In the initial materials and in the composting samples, pH and electrical conductivity (EC) were determined in the 1:10 (weight/volume) water soluble extract. The moisture content was analysed at 105°C for 24h. Organic matter (OM) was assessed by determining the loss-on ignition at 430°C for 24 h. Total organic carbon (Corg) was calculated according to the following equation C = OM/1.84 and total nitrogen (Nt) was determined by the Dumas method. After HNOivus L.) . Cation ribed by . Water-sribed by .All the 1) and final (X2) ash contents, with the The losses of OM were calculated using the initial  and carbohydrate molecules (at 200 to 420°C) during the combustion process.For the thermogravimetric analysis (TG), samples were air-dried, ground in an agate mill and sieved through a 0.125 mm mesh, and milled again with an agate mortar. Thermal analyses were performed with a TGA/SDTA851e/LF/1600 instrument  and Pfeiffer Vacuum (Thermostar GSD301T) mass spectrometer that enables the recording of thermograms and mass spectra of combustion gases simultaneously. All samples were combusted with a mixing stream of oxygen/He (20/80%), a gas flow 100 ml min2O5 and K2O) was determined. Then, the economic value of the nutrients present in the composts was assigned, considering a content of 25% of fresh matter in all the composts.The economic value of the nutrients included in the composts was calculated considering the price of diammonium phosphate (DAP), standard size, bulk, spot, free on board (f.o.b.) US Gulf; potassium chloride (muriate of potash), standard grade, spot, f.o.b. Vancouver and Urea; Black Sea, bulk, spot, f.o.b. Black Sea (primarily Yuzhnyy) beginning July 1991; for 1985–91 (June) f.o.b. Eastern Europe, provided by the World Bank. Based on these materials, the value of 100 kg of each respective compound . The normality and homogeneity of the variances were checked using the Shapiro-Wilk and Levene test, respectively, before ANOVA. The statistical analyses were conducted using the SPSS 20.0 software package.-1) and t the composting time (d). The RMS and F-values were calculated to compare the goodness of the curve fitting.OM losses during composting were fitted to a first-order kinetic function  by the M2 = 0.9445). This can be attributed to the higher content of easily-degradable organic compounds provided by TP or PP compared to MW. In piles P1 and P2, the temperature exceeded 55°C for more than two weeks, which ensured the maximum pathogen reduction according to the European guidelines on compost sanitation [In all the mixtures, the temperature values showed a rapid increase during the first days of composting due to the microbial proliferation, reaching the highest temperatures in the first week  . Other anitation . P3 did The initial concentrations of OM in the piles were 85.0% for P1, 84.9% for P2 and 95.7% for P3. The higher contents observed in P3 may be due to greater proportion of MW in this mixture; since this waste showed the higher concentrations of OM, together with TP . The OM OM degradation profile during composting followed a first-order kinetic equation in all the piles. Curve fitting of the experimental data gave the following parameter values (standard deviation in brackets):The highest values of A were observed for P3, related to the higher proportion of MW and PP. Similar behaviour were found for P1 and P2, according to the similar respective thermal profiles. Gavilanes-Terán et al.  reportedThermogravimetric analyses (TG) were carried out in the starting materials and in the composting samples to evaluate the chemical changes in the organic matter during the composting process. Thermolability is expressed by means of R1 index, defined as the ratio between the loss of mass in the range 420–550°C  in relation to the loss of mass in the range 200–420°C (due to carbohydrate molecules) during the combustion process . TherefoAs it has been commented, the first range of 200–420°C corresponds to the combustion of carbohydrates such as cellulose and lignocellulosic substances , 28, 31,The thermograms showed three main steps regions, defined by the different mass losses . The firThe R1 index increased during the composting process in all the piles . This inp-OH-benzoic, vanillic, veratric, syringic and p-coumaric acids [Regarding the evolution of the compost quality parameters during the process, pH values increased in all the mixtures during composting, obtaining only values close to neutrality in P3. The significant increase in the pH have been commonly found during composting of vegetable wastes , 27, 36,ic acids , and theic acids , 30.In relation to the salinity, the EC decreased in all the composting piles, showing lower values at the end of the process . EC usuaHowever, in this study, the composting process was developed in an open composting facility, which favours salt leaching by watering and especially, by rain. Other composting studies have reported this decrease in salinity due to salt leaching as a consequence of the rain, such as Gavilanes-Terán et al., during -1. The initial highest values were observed in P1 and P2, the composting piles elaborated with TP, which had the greatest contents in these compounds , which indicates a suitable degree of maturity in the composts obtained. The Corg/Nt ratio is also considered as an indicator of compost maturity and of the good development of the composting process [The cation exchange capacity (CEC) is another parameter strongly associated to the humification process that is usually used as indicator of these processes during composting . This pa process . At the  process . Due to  process . In thisCompost quality and in general, its value as organic fertilizer is still only determined by several standardized values  established in the different national and international legislations and/or guidelines.Regarding organic matter contents, all the composts practically fulfilled the requirements established in the different guidelines . In relaIn addition, the concentrations of K were similar to those obtained in composts from the agri-food industry  and in c2O5 and K2O could be averaged in 0.53, 2.27 and 0.55 €/100 kg. In addition, the economic value of the nutrients present in the composts has been calculated assigning a content of 75% of dry matter in all the composts. This demonstrates that the additional value of compost implies a value when its nutrient contents are considered, especially for the potassium value, representing around 70–75% of the total value of the evaluated macronutrients, involving cost savings.To assess the economic value associated to the compost nutrients, the average prices for urea, DAP, and potassium chloride in 2015 were considered according to the World Bank. These prices corresponded in euros/tonne to 241.83; 459.17 and 272.21 for urea, DAP, and potassium chloride respectively. Therefore, the value of the fertilizing units of N, PThe proposed strategies for the management and recycling of food market waste streams by co-composting with gardening pruning wastes have shown their feasibility in terms of organic matter mineralization and humification, obtaining composts with suitable characteristics for their agricultural use. All the composting mixtures showed a suitable development of the composting process, with a significant thermophilic phase, completed in a shorter period of time in pile 3. The composts obtained showed absence of phytotoxicity and suitable agronomic properties for their use as organic fertilizers. Therefore, considering all the environmental and economic issues related to waste management , it can be concluded that the proposed alternative should be scaled-up in developing areas to reduce and diversify the urban waste streams, producing high quality and balanced organic fertilizers, with a significant economic value in nutrients that could be internalized in the final compost price.S1 FileInclude data and figures corresponding to TG analyses of each composting pile. P1: 50% MW + 33% TP + 17% PP; P2: 60% MW + 30% TP + 10% PP; P3: 75% MW + 25% PP. MW: food market waste; TP: tree prunings; PP: palm prunings.(XLSX)Click here for additional data file.


Wilcoxon rank-sum test, Kruskal-Wallis analysis, Chi-squared test, and multiple logistic regression were employed to analyze the daily dietary vitamin C intake on the basis of three-day 24 h dietary recalls and food sources in relation to demographic factors, to evaluate vitamin C intake status using the estimated average requirement cut-off point, and to explore underlying influencing factors. The mean (SD (standard deviation)) and median (interquartile range) levels of the dietary vitamin C intake in adults were 78.1 (54.6) and 65.4 (61.4) mg/day, respectively. Light vegetables, dark vegetables, fruits, and tubers were the top four food sources, contributing a combined 97.3% of total daily dietary vitamin C intake in the study population. The prevalence of risk of insufficient dietary vitamin C intake was 65.1%. Both the distribution of vitamin C intake and the prevalence of risk of insufficient dietary vitamin C intake differed by several demographic factors. Educational level, residence area, geographic location, vegetable consumption, and total energy intake were independent determinants of the risk of insufficient dietary vitamin C intake. In conclusion, dietary vitamin C intake is inadequate in Chinese adult population, and an increase in vitamin C intake should be recommended especially to the population at risk for vitamin C insufficiency.Vitamin C is essential for human health. It is important to estimate the dietary vitamin C intake in the Chinese population to examine the effects of the nutritional transition occurred in recent decades. The present study aimed to estimate the dietary vitamin C intake in Chinese adults by using cross-sectional data from the 2015 China Nutritional Transition Cohort Study and selecting those aged 18–65 years with complete records of sociodemographic characteristics and dietary measurements ( Vitamin C, also known as ascorbic acid, is a water-soluble vitamin naturally present in some foods, particularly fruits and vegetables. It is used as a food additive and is available as a dietary supplement . BecauseAlong with the rapid economic growth and urbanization observed in China over the past few decades, the Chinese population has experienced a nutritional transition characterized by dramatic changes in the dietary patterns, eating, and cooking behaviors ,4. BetweAlthough fruits and vegetables remain the food groups with the highest contribution to total vitamin C intake ,8,9, sigTherefore, the present study aimed to estimate dietary vitamin C intake and vitamin C food sources and evaluate the prevalence of risk of inadequate vitamin C intake in the adult population using newly collected data from the China Nutritional Transition Cohort Study carried out in 2015. To identify independent determinants of the vitamin C intake status, this study further explored potential influencing factors. These findings would be valuable for developing and implementing public health strategies to meet the recommended vitamin C intake and provide guidance for population-specific nutrition education and intervention programs.Data in the present study were derived from the 2015 China Nutritional Transition Cohort Study conducted by National Institute for Nutrition and Health, Chinese Center for Disease Control and Prevention, which was an amplified survey based on the CHNS , an ongon = 56) and lactating (n = 108), and subjects with implausible energy intake . For participants with dietary consumption values outside the sex-specific limits of dietary consumption distribution  [n = 56 of <P1 and 56 of >P99 in males, n = 69 of <P1 and 69 of >P99 in females), fresh fruit consumption frequency , and fresh vegetable consumption frequency , we used the corresponding sex-specific P1 and P99 to replace those of <P1 and >P99, respectively. A total of 11,357 participants were involved in the analysis  by trained staff across three consecutive days, including two weekdays and one weekend day ,15.Based on the intake amount of each ingredient of foods consumed during the survey period, the vitamin C content of the reported food items was coded according to the Chinese Food Composition Table . Total vThe estimated vitamin C intake level was compared with that reported in the Chinese Dietary Reference Intakes , in whicFood items contributing to vitamin C intake were grouped according to the China Food Composition Table . Food soSimilarly to the dietary vitamin C estimation described in detail above, the energy content of each reported food item was coded according to the Chinese Food Composition Table on the basis of the individual dietary intake data during the survey period . The totThe participants were asked to complete a validated semi-quantitative food frequency questionnaire and report whether they consumed sixty-three foods and beverages during the last year and the frequency of that consumption, defined as the average number of times per day, week, month, or year, depending on whether the consumption was usual or unusual. The data on various fruit and vegetable consumption were used for the analysis. For each item, if the participant was a non-consumer, then his/her consumption frequency was set to zero time daily. For consumers, the reported average consumption frequency was converted into a uniform estimate of times/day by dividing times/week by 7, times/month by 30.5, and times/year by 365. Finally, the consumption frequency of each item was summed as that of fruits and vegetables, respectively, which was categorized into three levels  by tertiles.2; normal: 18.5 ≤ BMI < 24 kg/m2; overweight: 24 ≤ BMI < 28 kg/m2; and general obesity: BMI ≥ 28 kg/m2 [Height and body weight were measured by trained health workers on the basis of a standard protocol recommended by the World Health Organization . The heiQuestionnaires were employed to collect information for each subject in terms of age, gender, educational level, smoking and alcohol intake, household income, and residence region.n (%). Because of the non-normal distribution, the non-parametric Wilcoxon rank-sum test or the Kruskal-Wallis analysis were performed to test differences in the distribution of dietary vitamin C intake and the contribution percentage of food sources to the total vitamin C intake by subgroups of participants according to sociodemographic factors , moderate (≥2.4 and <10.9 thousand yuan/per capita) and high (≥10.9 thousand yuan/per capita) by tertiles; residence area: city, suburban, town or county capital city, and rural village classed according to the administrative divisions; geographic location: north or south depending on the Qinling Mountains–Huihe River Line, an important geographic boundary; fruit and vegetable consumption frequency, and BMI status). If the difference was significant among more than two groups, an additional multiple comparison was conducted by the Student-Newman-Keuls (SNK) method. The prevalence of risk of insufficient dietary vitamin C intake in adults and the proportion of subjects with a lower risk for insufficient dietary vitamin C by sociodemographic factors were compared by the Chi-squared test. Further, multiple logistic regression and quantile regression were employed to explore the differential and independent effects of the sociodemographic variables on the prevalence of risk of insufficient dietary vitamin C intake in adults and on various areas of vitamin C intake distribution, respectively. The dependent variables were the dietary vitamin C intake category (<85 vs. ≥85 mg/day) and the dietary vitamin C intake level in the logistic regression model and in the quantile regression model, respectively. On the basis of the results of the aformentioned analysis of the effects of sociodemographic factors on the distribution of dietary vitamin C intake and on the proportion of subjects at risk for insufficient dietary vitamin C intake, and of the findings in previous reports [p < 0.05 was considered significant. Data analysis was performed by SAS 9.4 . Continuous variables were presented as mean ± standard deviation (SD) and median (interquartile range), while categorical variables were expressed as  reports ,11,12,13A sample of 11,357 adults aged 18–65 years was examined in the present study; those aged 18–49 years accounted for 55.4% . About 5p < 0.05). The daily vitamin C intake was higher in males, in subjects from the southern part of China, and in those who drank alcohol than in their respective counterparts. Subjects with education at the middle school level  had larger vitamin C intake than those with primary school education or below , but no difference in vitamin C intake was found compared with those having a high school education or above . The daily vitamin C intake was higher in subjects from suburban areas than in those from any other residence areas. The daily dietary vitamin C intake was significantly different and presented an increasing trend associated with increased vegetable consumption frequency.The mean and median levels of dietary vitamin C intake in all subjects were 78.1 and 65.4 mg/day, respectively . Gender,p < 0.05). The prevalence of risk of inadequate vitamin C was greater, and the proportion of subjects with a lower likelihood of insufficient vitamin C was lower in females, in subjects living in northern China, and in those without alcohol intake, than in their respective counterparts. Among subgroups, the proportion of subjects at risk of insufficient dietary vitamin C seemed to be lower, and the proportion of subjects with less probability of insufficient dietary vitamin C intake was likely to be higher, in participants with middle school education, in those from suburban areas, in smokers, and in those with great vegetable consumption frequency. Moreover, the average levels of daily energy, protein, fat, and carbohydrate intake were 1940.9 kcal/day, 62.0 g/day, 77.0 g/day, and 245.4 g/day, respectively, in subjects with <85 mg/day of vitamin C intake, while those in subjects with ≥85 mg/day of vitamin C intake were 2273.3 kcal/day, 75.2 g/day, 92.4 g/day, and 279.5 g/day, respectively.Overall, the prevalence of risk of insufficient dietary vitamin C in the study population was 65.1%, and the proportion of subjects with a lower risk of insufficient dietary vitamin C intake was 25.9% , both ofp < 0.001, p < 0.001), but a partially lower fruit contribution percentage (p = 0.057) than their counterparts  , Polish The 15 provinces surveyed in the present study  vary subThis is a representative and large study evaluating the intake and food sources of vitamin C across sociodemographic factors and exploring independent determinants for vitamin C intake, using data of adults from 15 provinces in China. However, the findings cannot be extrapolated to the general population. Another limitation of this study is that the 24 h dietary recalls for three consecutive days were generally collected in summer, and seasonal variations in food consumption rich in vitamin C could not be taken into account. Moreover, there are always recall biases and a potential underestimation in dietary recalls being self-reported measurements. In addition, this study considers only intakes of vitamin C from dietary sources. Dietary supplements are likely to be considerable sources of vitamin C in some particular populations. It should be borne in mind that the users of supplements may differ from non-users with regard to demographic characteristics .Vitamin C intake in Chinese adults deviates from the recommended level, resulting in a high prevalence of risk of insufficient dietary vitamin C intake, which exerts potentially adverse influences on human health. Considering that fruit and vegetables are the main sources of vitamin C, the Chinese population should be encouraged to consume these foods according to dietary guidelines , especia


Therefore, molecules that can inhibit this enzyme are a source of potential anti-inflammatory drugs, with different action mechanisms of known anti-inflammatory agents. For the study and development of new anti-inflammatory drugs with this action mechanism, snake venom PLA2 (svPLA2) can be employed, since the svPLA2 has high similarity with the human PLA2 HGIIA. Despite the high similarity between these secretory PLA2s, it is still not clear if these toxins can really be employed as an experimental model to predict the interactions that occur with the human PLA2 HGIIA and its inhibitors. Thus, the present study aims to compare and evaluate, by means of theoretical calculations, docking and molecular dynamics simulations, as well as experimental studies, the interactions of human PLA2 HGIIA and two svPLA2s,Bothrops toxin II and Crotoxin B . Our theoretical findings corroborate experimental data and point out that the human PLA2 HGIIA and svPLA2 BthTX-II lead to similar interactions with the studied compounds. From our results, the svPLA2 BthTX-II can be used as an experimental model for the development of anti-inflammatory drugs for therapy in humans.Human phospholipase A Oxidation of AA generates inflammatory mediators, such as prostaglandins, thromboxanes and leukotrienes, through the action of cyclooxygenase (COX) and lipoxygenase (LOX) enzymes. In addition to AA, the breakdown of membrane phospholipids generates lysophospholipids, a precursor of platelet-activating factor (PAF), another potent inflammatory mediator oxy}propyl)phosphonic acid (PDB code U8D)) using the software Molegro Virtual Docker  [2. The best conformation was selected, based on the best overlap and the interaction energy. For the analysis of the svPLA2, the binding site, identified by the His47, was restricted into a sphere with 7 and 5 Å for the BthTX-II and CB, respectively, according to the size of the cavity. Since these enzyme structures do not have ligands, the best energy of interaction was taken into account. The selected conformations of all were used for the further MD simulation steps.To calculate the partial charges of ligands, the three-dimensional structures were previously created through the program PC SpartanCA, USA) , and theCA, USA) . The bin® program  [® 3.5 . The total energy, RMSD, and hydrogen bond graphs were generated to analyze the results through the Origin® program  [Initial ligand configurations were produced using the Gaussian 09 Program  to constIL, USA)  and DiscCA, USA) ,41,42,43


AbstractsubgenusArtibeus Leach, 1821 are widely distributed in Brazil. Conserved karyotypes characterize the group with identical diploid number and chromosome morphology. Recent studies suggested that the heterochromatin distribution and accumulation patterns can vary among species. In order to assess whether variation can also occur within species, we have analyzed the chromosomal distribution of constitutive heterochromatin in A.planirostris  and A.lituratus  from Central Amazon (North Brazil) and contrasted our findings with those reported for other localities in Brazil. In addition, Ag-NOR staining and FISH with 18S rDNA, telomeric, and LINE-1 probes were performed to assess the potential role that these different repetitive markers had in shaping the current architecture of heterochromatic regions. Both species presented interindividual variation of constitutive heterochromatin. In addition, in A.planirostris the centromeres of most chromosomes are enriched with LINE-1, colocated with pericentromeric heterochromatin blocks. Overall, our data indicate that amplification and differential distribution of the investigated repetitive DNAs might have played a significant role in shaping the chromosome architecture of the subgenusArtibeus.Species in the PageBreak Artibeus  are found in the Brazilian Amazon region: A.obscurus , A.lituratus , and A.planirostris   . These bogically . Neverthogically .subgenusArtibeus revealed a conserved karyotype, with diploid number (2n) of 30 chromosomes for females and 31 for males, with fundamental number, FNa = 56 , and proposed that the CH patterns were species-specific. However, although independent studies focused on species cytogenetic characterizations at the local level, no study has targeted the detection of interindividual CH variation in Artibeus within and among different Brazilian regions.Cytogenetic studies in all species of the FNa = 56 . The 2n is, 1856 . Althoug species . In theiin situ mapping of repetitive markers  can significantly contribute to the understanding of the evolution of genome architecture, as well as to the identification of intraspecific polymorphism in karyotypes otherwise conserved , as well as CH variation among representatives from Amazonian and other Brazilian regions. Furthermore, we have mapped rDNA and telomeric sequences on the karyotypes of both species to assess whether these sequences contribute to the architecture of centromeres and other positive heterochromatin blocks. As our final goal, we investigated the chromosomal distribution of LINE-1 sequences in A.planirostris chromosomes to i) compare with patterns described for other phyllostomid species, and ii) correlate the distribution of these sequences with the CH pattern observed for individuals in the same population.In the present study, we investigate whether there is CH variation within Central Amazon populations (North Brazil) of two Artibeus species used in this study are not listed as endangered at national or local levels. Our sampling included specimens of A.planirostris collected in an urban fragment at the National Institute of Amazonian Research (INPA) , and at “Bons Amigos” Farm . Furthermore, we collected individuals of A.lituratus at “Bons Amigos” Farm, Amazonas State, Brazil , followed by counterstaining with Propidium Iodide (0.2%) and mounting in Vectashield .Mitotic chromosomes were obtained from bone marrow cells using the o method . C-bandio method , the telethod  and fundamental numbers (FNa = 56), with 11 metacentric and three subtelocentric chromosome pairs (22m+6st+XX/XY1Y2). The X chromosome was a medium submetacentric, Y1 had a dot-like morphology, and Y2 was a small acrocentric , as well as in the distal region of short arms on three subtelocentric pairs , which are adjacent to the location of active Ag-NORs. The Y1 chromosome was euchromatic, and the Y2 had centromeric heterochromatin and additional blocks on the long arms. Likewise, the X chromosome showed centromeric heterochromatin and blocks on the short arms. The long arm of the X chromosome however, was not particularly enriched with heterochromatin  heterochromatic sites at centromeres, long arm, and distally on the short arm (A.lituratus from Pernambuco have two patterns: (i) centromeric, plus distal PageBreakon the short arm, and long arm  from the Amazon is too low to make generalizations, the similar number and location of heterochromatic blocks between individuals from Pará and Amazonas (both Northern Brazil), might indicate that specimens from the same ecogeographic regions have similar CH patterns. Pará and Amazonas states are contiguous and covered mostly by Amazon rainforest, whereas Pernambuco is a coastal state, separated from the Amazon by dry forests, and transitional environments, which might serve as mild dispersion barriers. Therefore, additional studies including large sampling are required to test the hypothesis that CH variation occurs by differential turnover of repetitive DNA , reinforced by geographical barriers through the distributional gradient of species.Although the number of analyzed individuals . The present Ag-NORs and 18S rDNA results agree with previously reported data , Urodermabilobatum W. Peters, 1866, U.magnirostrum Davis, 1968, Vampyriscusbidens , and Vampyressathyone O. Thomas, 1909  , and species in the same tribe as Artibeus  do not have multiple NORs (Artibeus (Artibeus NORs collocate (are adjacent) with heterochromatin and are likely associated with the amplification of heterochromatin in non centromeric regions.Silver nitrate staining and 18S rDNA FISH detected NORs at multiple sites on chromosomes of both species, more specifically distally on the short arms of the 5ted data . Many sps, 1856) . Howeverple NORs . TherefoArtibeus . As anotin situ hybridizations detected (TTAGGG)n telomeric sequences in all telomeres. Additionally, both species shared centromeric signals on three subtelocentric pairs : (i) these sequences might be telomere motifs reallocated from the terminal region of a chromosome to another chromosome or chromosome position; and (ii) the ITS presence on the centromere derives from reorganization of repetitive sequences (satellite DNA) composing these regions, which could also indicate the presence of centromeric hotspots of recombination during Artibeus karyotype evolution  and Phyllostomuselongatus   present ITS, which suggest lineage-specific events of amplification of these sequences can occur independently. Furthermore, it is noteworthy that pairs 5th–7th have ITS, cetromeric and non-centromeric CH blocks, as well as the NORs in all Artibeus specimens analyzed, suggesting that differential dynamics of heterochromatin DNA in these particular chromosomes might have played a role in the establishment of their shared distinct architecture, when compared to other autosomes. Refined investigation of these chromosomes at the sequence level will help disclosing whether differential heterochromatin composition contributed to the establishment of centromeric ITS.In both species, volution . The karhuffling . TherefoA.planirostris chromosomes revealed FISH signals near the centromere of most autosomes, except pairs 4, 7, 8, 13 and 14. , Lophostomaoccidentalis , and Gardnerycteriscrenulatum  . It was hypothesized that this unusual distribution might have contributed to the high degree of chromosomal reorganization in the genus Tonatia Gray, 1827. From our data, it is still premature to state that this retroelement or sequences derived from it are constitutional components of core centromeres. Similarly, because our probes comprised only a partial LINE-1 sequence, we cannot conclude that functional elements are contributing to the centromere dynamics PageBreakof Artibeus. Despite the uncertainty on what factors were responsible for the massive “colonization” of LINE-1s at centromeres, processes such as gene conversion, which promote homogenization of centromeric sequences are expected to facilitate the maintenance of LINE-1 sequences in high copy numbers in this region  . Centroms region .A.planirostris could be a result of amplification of different types of repeats (e.g. LINEs vs. satellite DNA) in specific chromosomes (We noticed inconsistent patterns when comparing the co-distribution of heterochromatin blocks and LINE-1 elements. Namely, in all analyzed individuals, interstitial CH blocks have LINE-1 signals in the second chromosome pair but not pair 1. Thus, non-centromeric heterochromatin formation on chromosomal arms of omosomes .1 and Y2 sex chromosomes presented weak FISH signals, contrasting with the strong signal throughout the long arm of the X (Fig. C.brevicauda chromosomes, which also presents a multiple sex chromosome system. They found that the original X chromosome had higher levels of LINE-1 accumulation than the translocated autosome. The X-autosome translocation of Artibeus is different from that observed in C.brevicauda, with the small autosome component representing the short arm of A.planirostris X chromosome. In agreement with A.planirostris had a significant accumulation of LINE-1, corresponding exactly to the original X chromosome. The LINE-1 accumulation on Y-chromosomes seems to be restricted to centromeric regions. In this case, we expect that similarly to other mammals, the original Y is mostly constituted of repeats other than retroelements, and that Y2 pattern corresponds to that observed for other autosomes due to its autosomal origin.The Ye X Fig. . LINE-1 e X Fig. . Parish Artibeus might have played a significant role in shaping the chromosome architecture of the genus, and we are still unsure if this trend at centromeres can be observed in other species of Stenodermatinae. Because the LINE-1 accumulation patterns differ in the bat species analyzed to date (Artibeus and PageBreakTonatia species, but not C.brevicauda, L.occidentalis, and G.crenulatum differred in the patterns of LINE-1 distribution.Transposable element activity and accumulation have been linked to chromosomal rearrangements and can be directly or indirectly associated with speciation events , 2014. I


Breast cancer is one of the most common malignancies in women. Patient-derived tumor xenograft (PDX) model is a cutting-edge approach for drug research on breast cancer. However, PDX still exhibits differences from original human tumors, thereby challenging the molecular understanding of tumorigenesis. In particular, gene expression changes after tissues are transplanted from human to mouse model. In this study, we propose a novel computational method by incorporating several machine learning algorithms, including Monte Carlo feature selection (MCFS), random forest (RF), and rough set-based rule learning, to identify genes with significant expression differences between PDX and original human tumors. First, 831 breast tumors, including 657 PDX and 174 human tumors, were collected. Based on MCFS and RF, 32 genes were then identified to be informative for the prediction of PDX and human tumors and can be used to construct a prediction model. The prediction model exhibits a Matthews coefficient correlation value of 0.777. Seven interpretable interactions within the informative gene were detected based on the rough set-based rule learning. Furthermore, the seven interpretable interactions can be well supported by previous experimental studies. Our study not only presents a method for identifying informative genes with differential expression but also provides insights into the mechanism through which gene expression changes after being transplanted from human tumor into mouse model. This work would be helpful for research and drug development for breast cancer. According to epidemiological data from the World Health Organization (WHO), breast cancer has been confirmed to be the most frequently diagnosed cancer in women from developed and developing countries and is accompanied with rising expectancy of average life span ,2. EarlyTo identify a cure for breast cancer, researchers have applied five levels of research, namely, molecular, cellular, histopathological, animal model, and clinical levels . Among tKRAS and PIK3CA, were greatly altered by the murine microenvironment across passages; hence, at the genomic level, the PDX mouse model cannot completely maintain the characteristics of in situ tumor tissues [Although the PDX mouse model maintains most of the characteristics of the original tumor, genomic and transcriptional variations induced by murine microenvironment selection have been characterized. In 2014, researchers from University of North Carolina (USA) found that in pancreatic and colorectal cancer, the mutation frequencies of two oncogenic genes, namely,  tissues . At the  tissues .In this study, we obtained and used the gene expression data from a recent study  on PDX mOur entire data set contained 831 tumor cells, including 657 PDX and 174 tumor cells. The 831 tumor cells were encoded into feature vectors by the expression levels of 69 genes. The 69 genes we used were derived from the work by Lawson et al. . To inveOur method for identifying differentially expressed genes consisted of three steps: (i) applying MCFS method ,15,16 toThe expression levels of 116 genes in 1293 tumor cells from three patients with breast cancer and the corresponding PDX models were obtained from a recent publication . These gRI) to a feature if this feature is selected by more tree classifiers. Suppose the total of s×t classification trees are constructed by selecting s subsets of m features , and constructing t trees for each of the s subsets. Each of the t trees is built and trained by a random selection of training and test sets using the original training set. The RI of a feature can be measured by estimating the overall number of splits involving the feature in all nodes of all trees constructed. Particularly, the RI of a feature g can be estimated by the following equationMCFS method is designed to select informative features for supervised classifiers by randomly constructing a large number of tree classifiers from an original training dataset. MCFS assigns higher relative importance . In this study, we performed MCFS ranking by using the MCFS software package . MCFS prF. However, determining the optimal number of features to be used is difficult. Therefore, IFS [We used MCFS method to evaluate the importance of the 69 features, resulting in a feature list ore, IFS ,23,24 anore, IFS ,26 were F, F, and so on. To select the optimal feature set, i.e., determining the optimal features to be used, RF classification algorithm was then performed on all candidate feature sets. The classification performance of each candidate model was evaluated by cross-validation [IFS method first constructed a series of candidate feature sets, denoted as lidation ,28  algorithm which was used to generate decision rules based on rough set theory.After obtaining the informative genes, the rough set-based rule learning algorithm can be applied to detect interaction between informative genes. The detected interactions were represented as rules. A rule is a terminology to describe a relation between rule conditions (the left-hand-side of the rule) and the rule outcome (the right-hand-side). For example, in our study, a rule can be presented as IF–THEN relationship based on gene expression: IF Gene1 = Rough set is described briefly as follows. A decision (classification) system ifferent . A functThe entry A Boolean product-of-sums (POS) function, called discernibility function ough set . A disceAccordingly, the reducts sorption . Howeverlgorithm  is appliJohnson Reducer , which iBased on the reduct, the RIPPER algorithm is applied to generate decision rules. RIPPER, proposed by Cohen  in 1995,In our study, we fulfilled the detection of rules by using the Johnson Reducer algorithm , which wTenfold cross-validation ,44,45,46n samples, denoted by N classes, represented by 1, 2, …, N. True classes of samples were defined as matrix j; otherwise, this variable is set to 0. Predicted classes of samples were defined as matrix j; otherwise, X,Y) was covariance function of X and Y, which can be computed bykth column of X and Y, respectively; Given the involved MCC is between −1 and 1. With high MCC value, the classifier yields a good performance . The range of RI on the tumor cell classification. Then, by applying IFS and RF methods, we drew the IFS-curve, which illustrated the relationship between the number of features used for building the model and the corresponding performance measured by the MCC value  and regarded as the biomarker of cancer cell stemness [MUC1 can be altered in this progress. Similarly, another gene involved in the stemness maintenance of cancer cells, PROM1 [In encoding a specific membrane glycoprotein, stemness ,61,62. As, PROM1 ,63, can CXCR4 encodes a specific G protein coupled receptor, which has been widely reported to be expressed on the immature CD34+ hematopoietic stem cells [CXCR4 are very diverse among the primary tumor of breast cancer, the metastatic tumor tissues, and the PDX tumors filtered by specific murine microenvironment [em cells ,65. Receironment ,67. ERBB2 and ERBB3, have been predicted as potential distinctive biomarkers. ERBB family signaling pathway has been widely reported to contribute to transcriptional regulation during the tumorigenesis of breast cancer [ERBB2 and ERBB3 have been systematically regulated for further adaption to the murine mouse microenvironment [In our results, two specific Erb-B2 Receptor Tyrosine Kinase (ERBB)-pathway-associated genes, i.e., t cancer . Furtherironment ,69.ID4, has also been identified as a breast-cancer-associated gene, which participates in the regulation of basic helix-loop-helix transcription factors [ID4 may also be changed in a different microenvironment.Another gene,  factors ,71. Alth factors ,73. As tPTEN, NTRK2, PGR, and TP53, which are identified in this study as the differentially expressed genes.A recent study on single-cell transcriptome of PDX mouse model reported a series of genes with differential expression patterns in the PDX mouse model compared with the original tumors ; this seKRT19, EMP1, CXCR4, CD44, PTEN, PARP2, and PLCB4, were required to have high expression levels in the original tumor cells, whereas others should have low expression levels. As we discussed above, some of these genes, such as KRT19, EMP1, and PARP2, have evidence from previous studies, which indicate their higher expression levels in the original tumors compared with PDX tumor cells [As we have mentioned above, we detected seven specific rules to distinguish primary tumor tissues and PDX tumor tissues based on the expression levels of 14 genes. In the seven rules, the first six provided criteria for classifying a tumor cell into the original human tumor, which also represents the primary tumor-specific gene expression patterns see . Among tor cells ,13,52,58KRT19, KRT5 and CDH3 (interactions of dark red color). Thus, we provided full analysis on the interactions among KRT19, KRT5 and CDH3.Further, to facilitate our understanding of the seven rules, we visualized the seven rules by using Ciruvis, a web-based tool for rule networks and interaction detection using rule-based classifiers, as suggested in the literature . The resKRT19 and KRT5 has been indicated by previous experiments. First, the dimerizing of Krt5 with Krt14 has been confirmed to be involved in the regulation of EMT in tumorigenesis ,76,77. FKRT5 and CDH3, which encodes a member of the cadherin superfamily. A previous study in 2010 [BRCA1 repressed the expression of various cytokeratins including Krt5. It is also known that BRCA1 product manipulate the expression pattern of both KRT5 and CDH3 [Second, the Ciruvis figure also show strong correlations between  in 2010  reportedand CDH3 , suggestKRT19 and CDH3 is suggested by the previous studies which reported the participation of these two genes in the essential proliferation and metastasis associated pathway MAPK/ERK cascade in tumorigenesis [KRT19 shows co-expression pattern with various CDH3 associated genes like CDH1 [EGFR [CTNNB1 [KRT19 and CDH3. Third, the correlation of igenesis ,83. Furtike CDH1 , EGFR [8H1 [EGFR  and CTNN [CTNNB1  in certaKRT19, KRT5 and CDH3, which are all related to EMT associated pathological processes in human tumors, but not in PDX mouse model.In summary, these findings strongly support the correlations of both functions and expression patterns among the three genes Through a new computational method, we identified the differentially expressed genes and quantitative rules that can classify the PDX tumor and the original tumor with high accuracies. Multiple well-known oncogenes and tumor suppressors are differentially expressed; this finding indicates the risk of using those genes as therapeutic targets in breast cancer through PDX mouse model. Therefore, our study not only provides a functional tool to distinguish the primary tumor from PDX tumor by gene expression levels but also reveals the detailed gene expression alterations and shaping characteristics in murine microenvironment. In this work, we employed SVM, RF, dagging and rough set classifiers due to the small samples problem. Some of the latest classifiers, such as ensemble classifier , will be