The secretome from human-derived mesenchymal stem cells augments the activity of antitumor plant extracts in vitro.

Cancer is understood as a multifactorial disease that involve multiple cell types and phenotypes in the tumor microenvironment (TME). The components of the TME can interact directly or via soluble factors (cytokines, chemokines, growth factors, extracellular vesicles, etc.). Among the cells composing the TME, mesenchymal stem cells (MSCs) appear as a population with debated properties since it has been seen that they can both promote or attenuate tumor progression. For various authors, the main mechanism of interaction of MSCs is through their secretome, the set of molecules secreted into the extracellular milieu, recruiting, and influencing the behavior of other cells in inflammatory environments where they normally reside, such as wounds and tumors. Natural products have been studied as possible cancer treatments, appealing to synergisms between the molecules in their composition; thus, extracts obtained from Petiveria alliacea (Anamu-SC) and Caesalpinia spinosa (P2Et) have been produced and studied previously on different models, showing promising results. The effect of plant extracts on the MSC secretome has been poorly studied, especially in the context of the TME. Here, we studied the effect of Anamu-SC and P2Et extracts in the human adipose-derived MSC (hAMSC)-tumor cell interaction as a TME model. We also investigated the influence of the hAMSC secretome, in combination with these natural products, on tumor cell hallmarks such as viability, clonogenicity, and migration. In addition, hAMSC gene expression and protein synthesis were evaluated for some key factors in tumor progression in the presence of the extracts by reverse transcription-quantitative polymerase chain reaction (RT-qPCR) and Multiplex, respectively. It was found that the presence of the hAMSC secretome did not affect the cytotoxic or clonogenicity-reducing activities of the natural extracts on cancer cells, and even this secretome can inhibit the migration of these tumor cells, in addition to the fact that the profile of molecules can be modified by natural products. Overall, our findings demonstrate that hAMSC secretome participation in TME interactions can favor the antitumor activities of natural products.

Induced selectivity in the photochemistry of estrone derivatives in sustainable and micellar environment: preparative and mechanistic studies.

In this study, we carried out preparative and mechanistic studies on the photochemical reaction of a series of 3-acylestrone derivatives in confined and sustainable micellar environment under steady-state conditions and the results were compared with those obtained in cyclohexane solution. The aim of this work is mainly focused to show whether the nature of the surfactant (cationic, neutral and anionic) leads to noticeable selectivity in the photoproduct formation. The 3-acylestrone derivatives underwent the photo-Fries rearrangement, with concomitant homolytic fragmentation of the ester group and [1;3]-acyl migration. This pathway afforded the ortho-acyl estrone derivatives, the main photoproducts together with estrone. However, epimerization of the ortho regioisomer 2-acetylestrone and estrone through Norrish Type I photoreaction occurred involving the fragmentation of the C-α at the carbonyl group (C-17) of the steroid. UV-visible and 2D-NMR (NOESY) spectroscopies have been employed to measure the binding constant Kb and the location of the steroids within the hydrophobic core of the micelle.

Clinical and endoscopic features in patients with hospital-acquired diarrhea associated with Clostridium difficile infection. / Características clínicas y endoscópicas en diarrea hospitalaria asociada a infección por Clostridium difficile.

INTRODUCTION AND AIMS: Clostridium difficile infection is the main cause of hospital-acquired diarrhea, and the clinical and endoscopic findings in those patients have been studied very little in Mexico. The aim of the present study was to describe those findings. MATERIALS AND METHODS: A prospective cohort study was conducted that included patients with hospital-acquired diarrhea associated with Clostridium difficile diagnosed through polymerase chain reaction. The hypervirulent NAP027 strain was also determined. The clinical and endoscopic findings in the study patients, as well as the variables associated with severity, were analyzed. RESULTS: Of the 127 patients with hospital-acquired diarrhea, 97 were excluded from the study due to lack of colonoscopy. The remaining 39 study patients had a mean age of 48 years, and their most common signs/symptoms were abdominal pain (49%), mucus in stools (41%), and blood in stools (10%). The most common alterations in the laboratory results were leukocytosis in 49%, fecal leukocytes (61%), and hypoalbuminemia (67%). The main risk factor was antibiotic use in 62%, and ceftriaxone was the most widely used. The hypervirulent strain was present in 54% of the cases. Endoscopic abnormalities were found in 87% of the patients. Thirty-eight percent presented with pseudomembranous colitis, with lesions in the left colon in 53%, and in the right colon in 13%. No association was found between proton-pump inhibitor use and Clostridium difficile-associated diarrhea. There was a significant association between hypoalbuminemia (< 3.3g/dL) and a greater risk for severe colitis, with a RR of 8.2 (p=0.008). CONCLUSIONS: Pseudomembranous colitis lesions associated with the hypervirulent Clostridium difficile strain were predominant in the left colon. Hypoalbuminemia was a significant severity predictor.

Precooling treatments induce resistance of Anastrepha ludens eggs to quarantine treatments of high-pressure processing combined with cold.

High-pressure processing (HPP) combined with heat or cold has been proposed as an alternative quarantine process for Mexican fruit fly Anastrepha ludens Loew (Diptera: Tephritidae). HPP conditions at levels higher than 100 MPa applied to destroy eggs and larvae can also affect the postharvest physiology of the fruits. HPP at pressure levels in the range of 50-100 MPa is recommended. Eggs have been reported as being more resistant to HPP than larvae. Therefore, the objective of this study was to assess the effect of a precooling treatment on the biological viability of A. ludens eggs treated by HPP at 0 degrees C. The capability of nondestroyed eggs to develop and reproduce was also evaluated. One-, 2-, 3-, and 4-d-old eggs were precooled in ice water for 0 (control) 3, 6, 12, or 24 h and then pressurized at 50, 70, or 90 MPa for 0, 3, 6, or 9 min at 0 degrees C. The hatching capability of pressurized eggs was evaluated. The most lethal effect of HPP on nonprecooled eggs (0 h) was obtained at 90 MPa for 9 min, destroying all eggs except for the 3-d-old ones, which showed an 11.8% hatch rate. Precooling treatment improved the hatch rate of eggs ranging from 4 to 50% depending on precooling conditions. The main effect was observed after 6 h. These results suggest that precooling modified the biochemistry and physiology of eggs, improving their resistance to HPP treatments.

Influence of taste sensitivity on preference and sensory perception of mezcal.

The perception of orosensory stimuli, which includes flavor, can vary between individuals. These individual variations in oral sensations can be due to genetic factors and it would appear that they can predict food liking and consumption. The most studied source of variation is related to bitter taste perception associated with 6-n-propylthiouracil (PROP) responsiveness. In this context, humans can be classified as non-tasters (NT), medium tasters (MT) and supertasters (ST). Evidence suggests that genetic variation in bitter taste perception contributes to differences in the level of irritation caused by alcohol perception in solutions. The aim of this investigation was to study the bitter taste sensitivity among a group of mezcal consumers and its relationship with sensory perception and preference through PROP taster status. The tests were carried out in the state of Oaxaca in Mexico. A total of 83 mezcal consumers were classified by their PROP taster status and were asked to provide sensory descriptors for five mezcal samples and rate them according to the level of liking. The three-solution test was used to classify the subjects as NT, MT, and ST, while a Multiple Factor Analysis (MFA) was used to visualize the sensory descriptors provided by these three groups. The proportion of MT subjects was 16%, while the proportion of NT and ST was 34 and 51%, respectively. The MT provided higher liking ratings for at least three mezcal samples. According to MFA, the mezcal samples were organized in a similar configuration along the two dimensions. However, NT mentioned a limited number of simple terms (strong flavor, tasteless, burning in the mouth) to describe the samples, whereas ST used a more complex vocabulary (astringent, smoky, scratchy aftertaste). These data suggest that the preference for mezcal samples was similar for non-taster and supertasters, but there are indications that the sensory perception of mezcal differs between groups.

Empiric therapy directed against MRSA in patients admitted to the intensive care unit does not improve outcomes in community-acquired pneumonia.

PURPOSE: The Infectious Diseases Society of America has recommended empiric therapy active against methicillin-resistant Staphylococcus aureus (MRSA) for all community-acquired pneumonia (CAP) patients admitted to the intensive care unit (ICU). However, there is sparse data to support this recommendation. The objective of our study was to ascertain if such a practice improves outcomes. METHODS: This study was a secondary, retrospective analysis of the Community-Acquired Pneumonia Organization (CAPO) international database on CAP. Outcomes in patients admitted to the ICU were compared according to empiric initiation of anti-MRSA therapy (vancomycin or linezolid) with standard ICU CAP therapy (MRSA therapy group) or standard therapy alone for ICU CAP (standard therapy group). RESULTS: A total of 621 patients were identified with ICU pneumonia, of whom 57 patients had been initiated empirically on vancomycin or linezolid (MRSA therapy group). Patients of the MRSA therapy group had more comorbidities and were more severely ill than those of the standard therapy group. However, there were no statistical differences between the MRSA therapy group and standard therapy group for the primary outcomes of in-hospital and 28-day mortality, length of stay and time to clinical stability. CONCLUSIONS: These findings suggest that empiric MRSA therapy in all ICU CAP patients may not improve outcomes and argue for clinician review of local epidemiologic trends on MRSA prevalence to ascertain the need for empiric MRSA coverage.

Effect of high hydrostatic pressure on the physiology of Manila mango.

Manila mangoes (Mangifera indica L.) have sensory characteristics that make them attractive for consumption as a fresh fruit. A large portion of the annual yield of this fruit is infested by the Mexican fruit fly (Anastrepha ludens), adversely impacting the quality of the crop. Hence, it is necessary to develop economically viable postharvest treatments to reduce the damage caused by this insect. Currently, high hydrostatic pressures are used to guarantee the safety of many processed foods. The objective of this work was to assess the effects of high hydrostatic pressure on mangoes at their physiological maturity. High hydrostatic pressures were applied to mangoes at three levels: 50, 100 and 200 megapascals applied for four different time periods (0, 5, 10 and 20 min). Physiologically mature mangoes were more resistant to changes in response to the pressure of 50 MPa. Reduction of physiological activity by application of high hydrostatic pressure opens a new avenue for the research on treatments intended to enhance preservation of whole fresh fruit.

An Hcp100 gene fragment reveals Histoplasma capsulatum presence in lungs of Tadarida brasiliensis migratory bats.

Histoplasma capsulatum was sampled in lungs from 87 migratory Tadarida brasiliensis bats captured in Mexico (n=66) and Argentina (n=21). The fungus was screened by nested-PCR using a sensitive and specific Hcp100 gene fragment. This molecular marker was detected in 81·6% [95% confidence interval (CI) 73·4-89·7] of all bats, representing 71 amplified bat lung DNA samples. Data showed a T. brasiliensis infection rate of 78·8% (95% CI 68·9-88·7) in bats captured in Mexico and of 90·4% (95% CI 75·2-100) in those captured in Argentina. Similarity with the H. capsulatum sequence of a reference strain (G-217B) was observed in 71 Hcp100 sequences, which supports the fungal findings. Based on the neighbour-joining and maximum parsimony Hcp100 sequence analyses, a high level of similarity was found in most Mexican and all Argentinean bat lung samples. Despite the fact that 81·6% of the infections were molecularly evidenced, only three H. capsulatum isolates were cultured from all samples tested, suggesting a low fungal burden in lung tissues that did not favour fungal isolation. This study also highlighted the importance of using different tools for the understanding of histoplasmosis epidemiology, since it supports the presence of H. capsulatum in T. brasiliensis migratory bats from Mexico and Argentina, thus contributing new evidence to the knowledge of the environmental distribution of this fungus in the Americas.

Computed tomography measurements of parapneumonic effusion indicative of thoracentesis.

Patients with parapneumonic effusions (PPE) measuring <1 cm by lateral decubitus radiograph (LDR) or <5 cm by lateral erect radiograph (LER) do not require thoracentesis. No such data exist for chest computed tomography (CCT). The objective of this study was to identify a PPE measurement by CCT that indicates the need for thoracentesis. A secondary data analysis of two pneumonia databases was conducted to identify patients with PPE. Measurements of PPE using LDR, LER and CCT were correlated by linear regression analysis. The clinical outcome of community-acquired pneumonia patients managed with the newly defined CCT measurement was evaluated. PPE was identified in 419 out of 1,460 patients with possible pneumonia. PPE measurements of 1 cm and 5 cm by LDR and LER, respectively, correlated with a measurement of 2.5 cm by CCT. Out of 95 patients with CCT measurements <2.5 cm, 31 poor clinical outcomes were reported: outcome was PPE related (n = 1); outcome was PPE unrelated (n = 26); and outcome was not evaluable (n = 4). The single case of poor outcome also measured <1 cm by LDR. This study indicates that patients with community-acquired pneumonia and a PPE measuring <2.5 cm by CCT can be managed without the need for thoracentesis.

Duration of antibiotic therapy in hospitalised patients with community-acquired pneumonia.

Recent guidelines suggest that duration of antibiotic therapy for hospitalized patients with community-acquired pneumonia (CAP) can be reduced by individualising treatment based on patient's clinical response. However, the degree of application of this principle in clinical practice is unknown. Duration of therapy was analysed in patients identified from the Community-Acquired Pneumonia Organization database and evaluated with respect to severity of the disease on admission and time to clinical stability (TCS). Among the 2,003 patients enrolled, mean duration of total antibiotic therapy was 11 days. Neither the pneumonia severity index (r(2) = 0.005) nor the CRB-65 (r(2) = 0.004) scores were related to total duration of therapy. Duration of intravenous antibiotic therapy was related to TCS (r(2) = 0.198). Conversely, TCS was not related to duration of either oral (r(2) = 0.014) or total (r(2) = 0.02) antibiotic therapy. Neither TCS nor other characteristics were found to be significantly associated with duration of total therapy by logistic regression analysis (r(2)<0.09). The individualised approach suggested by recent guidelines has not been adopted in current clinical practice. Duration of therapy is not influenced by either the severity of disease at the time of hospitalisation or the clinical response to therapy.

Hospitalization due to community-acquired pneumonia in patients with chronic obstructive pulmonary disease: incidence, epidemiology and outcomes.

OBJECTIVES: Community-acquired pneumonia (CAP) is an important complication in patients with chronic obstructive pulmonary disease (COPD). This study aimed to define incidence, and outcomes of COPD patients hospitalized with pneumonia in the city of Louisville, and to estimate the burden of disease in the US population. METHODS: This was a secondary analysis of a prospective population-based cohort study of residents in Louisville, Kentucky, 40 years old and older, from 1 June 2014 to 31 May 2016. All adults hospitalized with CAP were enrolled. The annual incidence of pneumonia in COPD patients in Louisville was calculated and the total number of adults with COPD hospitalized in the United States was estimated. Clinical outcomes included time to clinical stability (TCS), length of hospital stay (LOS) and mortality. RESULTS: From a Louisville population of 18 246 patients with COPD, 3419 pneumonia hospitalizations were documented during the 2-year study. The annual incidence was 9369 patients with pneumonia per 100 000 COPD population, corresponding to an estimated 506 953 adults with COPD hospitalized due to pneumonia in the United States. The incidence of CAP in patients without COPD was 509 (95% CI 485-533) per 100 000. COPD patients had a median (interquartile range) TCS and LOS of 2 (1-4) and 5 (3-9) days respectively. The mortality of COPD patients during hospitalization, at 30 days, 6 months and 1 year was 193 of 3419 (5.6%), 400 of 3374 (11.9%), 816 of 3363 (24.3%) and 1104 of 3349 (33.0%), respectively. CONCLUSIONS: There was an annual incidence of 9369 cases of hospitalized CAP per 100 000 COPD patients in the city of Louisville. This was an approximately 18-fold greater incidence of CAP in COPD patients than in those without COPD.

The role of neutropenia on outcomes of cancer patients with community-acquired pneumonia.

Although the presence of neutropenia may predispose cancer patients to develop community-acquired pneumonia, the role of neutropenia on their outcomes has not been investigated. The purpose of the present study was to compare clinical outcomes of cancer community-acquired pneumonia patients with and without neutropenia. Patients with cancer, identified in the Community-Acquired Pneumonia Organization database, were divided into two groups according to the type of cancer and the presence of neutropenia: patients with solid cancer without neutropenia versus those with functional or absolute neutropenia. Among the 3,106 community-acquired pneumonia patients enrolled, 135 had cancer without neutropenia and 75 had cancer with neutropenia. No significant difference was found between patients with and without neutropenia regarding mean time to clinical stability (5.4+/-2.7 versus 4.9+/-2.7 days, respectively), mean length of hospital stay (9.2+/-7.7 versus 9.9+/-9.6 days) and in-hospital mortality (18 versus 15%, respectively). Using a multiple logistic regression model, neutropenia was not associated with mortality in cancer patients when adjusting for significant covariates (odds ratio 1.30). Lack of neutropenia, during the initial evaluation of a cancer community-acquired pneumonia patient, should not be considered an indicator of better clinical outcome.

Paradoxical sleep in two species of avian predator (Falconiformes).

Periods of disconjugate. and conjugate eye movements occur during the sleep cycle in Buteo jamaicensis arborealis and Herpetotheres cachinnans chapmanni. Electromyograms are essentially isoelectric throughout sleep. Slow waves appear only in short bursts of low amplitude in contrast to the long trains of high-amplitude waves reported for chickens and pigeons.

Mechanical and functional properties of beef products obtained using microbial transglutaminase with treatments of pre-heating followed by cold binding.

Beef proteins are considered non-setting proteins and usually gels obtained by adding of microbial transglutaminase are obtained by cooking directly the solubilized paste. The aim of this work was to determine the effect of pre-heating treatments on the mechanical properties of restructured beef gels treated with microbial transglutaminase (MTG). The effect of cooling (cold binding) the solubilized pastes after the pre-heating treatments was also studied. The restructured beef gels were obtained by adding 0.3% MTG or 0% MTG (control). Three pre-heating temperatures (40, 50 or 60°C) for 30 or 60min were studied, followed by heating at 90°C for 15min. Control samples without pre-heating were also prepared. Cold binding was studied by holding pre-heated gels at 4°C for 12h before heating at 90°C for 15min. Changes in mechanical properties (texture profile analysis and puncture test), color attributes, expressible water and cooking loss were determined. Results indicated that the better mechanical properties can be obtained by pre-heating beef pastes at 50°C for 30min with minimal effect on color, expressible water and cooking loss when 0.3% of MTG is added. It was concluded that there were no practical advantages by pre-heating the gels for 60min. Cold binding did not improve the mechanical properties of beef gels.

Fractionation of U and heavy metals into the colloidal fraction in acid mine drainage conditions in the Río Tinto area (SW Spain).

The Río Tinto mining area provides ideal conditions for studying the role of colloidal particles in concentrating. All the elements are present in higher concentrations respect to their content in common natural waters. 234U/238U activity ratio ranges between 2.286 ±â€¯0.149 and 2.531 ±â€¯0.151. Concentration of Fe and Al are in the order of 104-103 mg/L; Co, Zn and Cu values are three times lower than Fe and Al, but still much greater than in natural waters; U reaches values up to 40.73 µg/L. To evaluate the partitioning of these elements into the colloidal fraction, ultrafiltration (UF) experiments were made. A permeation model describing the relationship between concentration of the elements or isotopes in the permeate solution and the concentration factor (CF) was calculated. Experiments were carried out using CF from 1.5 to 25.0. The behaviour of investigated heavy metals and U can be predicted by a permeation model that provides similar permeation coefficient (Pc) for all of them (Pc = 0.95-0.97 for 50 kDa, Pc = 0.86-0.88 for 10 kDa, and Pc = 0.80-0.86 for 3 kDa). Retention for all the investigated elements occurs, resulting in up to 23% of the element associated to the 3 kDa colloidal fraction. The increase in U and heavy metals concentrations in the retentate fraction with the increasing CF is due to the retention of the colloidal fraction. The colloidal abundance of related metals increased with decreasing membrane MWCO even though the difference is small between 3 and 10 kDa fraction for some metals Characterization of colloidal material is needed to better understand the state of play in this context.

Colorectal cancer survival at an oncologic center in Colombia. A historic cohort study. / Supervivencia de cáncer colorrectal en un centro oncológico de Colombia. Estudio de cohorte histórica.

INTRODUCTION AND AIMS: In Colombia, cancer of the colon is the third most frequent cancer in relation to incidence and mortality. Five-year survival depends on stage at diagnosis, albeit that rate is not known for the country. The aim of the present study was to characterize the overall survival and disease-free survival rates in an adult population with colorectal cancer treated at an oncology center in Medellín, Colombia. MATERIALS AND METHODS: A retrospective cohort study was conducted. The case records of patients with a histologic diagnosis of colorectal cancer, seen within the time frame of 2011 and 2015, were reviewed. The overall survival and disease-free survival curves were calculated using the Kaplan-Meier method. RESULTS: A total of 824 (54.9%) patients with cancer of the colon and 676 (45.1%) with cancer of the rectum were treated. Mean patient age was 63.3 years, female sex predominated (56.3%), and 98.1% of the tumors were adenocarcinomas. The majority of the lesions were stage iii (31.9% in the colon and 35.5% in the rectum) at the time of diagnosis. Surgery was the most frequent treatment in the colon (85.2%) and radiotherapy was the most frequent in the rectum (75.4%). Overall survival at the median follow-up (27.3 months) was 66.7% for cancer of the colon and 63.9% for cancer of the rectum. Disease-free survival at the median follow-up (18.6 months in colon and 14.9 in rectum) was 72.5 and 68.9%, respectively. CONCLUSIONS: The clinical characteristics and treatment of patients were similar to those found in other studies. Two-year survival was higher than in other Colombian reports and 5-year survival was lower than that observed in developed countries.

Fostering international multicenter collaborative research: the CAPO Project.

The Community-Acquired Pneumonia Organization (CAPO) was created in 1999, with the goal to facilitate international research in the field of pneumonia. This manuscript reviews the development of CAPO, summarises the core clinical study and functions of the project's statistical and data coordinating center and describes current research opportunities. Investigators may be interested in joining the CAPO research team or applying the CAPO model for international collaboration to other areas of clinical research.

Association of serum pepsinogen with atrophic body gastritis in Costa Rica.

Individuals with atrophic gastritis (AG), especially atrophic body gastritis (ABG), are at increased risk of developing gastric cancer. Serum concentrations of pepsinogens (PG) have been proposed as markers for ABG. The aim of this study was to determine the risk factors for AG and ABG and the potential of using serum PG concentrations to detect ABG in a dyspeptic population in Costa Rica, which is one of the countries with the highest incidence and mortality rates of gastric cancer in the world. Seven biopsy specimens, a fasting blood sample and a questionnaire concerning sociodemographic factors were obtained from 501 consecutive dyspeptic patients. The serum PGI level and the PGI/PGII ratios were significantly lower in patients with ABG than in other groups (P<0.000). A cut-off point of 3.4 led to a sensitivity of 91.2% in identifying ABG, a negative predictive value of 98.1%, but a positive predictive value of only 11.2%. Helicobacter pylori were present in 93% of the patients and all those with peptic ulcers were positive. AG was associated with increased age, lower body mass index, high alcohol intake and low fruit consumption. ABG was associated with age, alcohol consumption and PGI/PGII<3.4. In dyspeptic patients with a high prevalence of H. pylori infection, serum PG levels provide an assessment of ABG but it is necessary to introduce other serological and genetic markers in order to achieve a better specificity. Those markers could be serum antibodies to H. pylori-CagA, cytokine gene polymorphisms or others.

Hyperhomocystinemia in patients with coronary artery disease.

Hyperhomocystinemia has been related to an increased risk of cardiovascular disease in several studies. The C677T polymorphism for the gene that encodes the methylenetetrahydrofolate reductase enzyme (MTHFR) and low plasma folate levels are common causes of hyperhomocystinemia. Due to differences in nutritional patterns and genetic background among different countries, we evaluated the role of hyperhomocystinemia as a coronary artery disease (CAD) risk factor in a Brazilian population. The relation between homocysteine (Hcy) and the extent of CAD, measured by an angiographic score, was determined. A total of 236 patients referred for coronary angiography for clinical reasons were included. CAD was found in 148 (62.7%) patients and 88 subjects had normal or near normal arteries. Patients with CAD had higher Hcy levels [mean (SD)] than those without disease (14 (6.8) vs 12.5 (4.0) microM; P = 0.04). Hyperhomocystinemia (Hcy >17.8 microM) prevalence was higher in the CAD group: 31.1 vs 12.2% (P = 0.01). After adjustment for major risk factors, we found an independent association between hyperhomocystinemia and CAD (OR = 2.48; 95% CI = 1.02-6.14). Patients with a more advanced coronary score had a higher frequency of hyperhomocystinemia and tended to have higher mean Hcy levels. An inverse relation between plasma folate and Hcy levels was found (r = -0.14; P = 0.04). Individuals with the MTHFR C677T polymorphism had a higher prevalence of hyperhomocystinemia than those without the mutated allele. We conclude that hyperhomocystinemia is independently associated with CAD, with a positive association between Hcy level and disease severity.