Bacteroidales as a fecal contamination indicator in fresh produce industry: A baseline measurement.

Foodborne outbreaks caused by fecal contamination of fresh produce represent a serious concern to public health and the economy. As the consumption of fresh produce increases, public health officials and organizations have pushed for improvements in food safety procedures and environmental assessments to reduce the risk of contamination. Visual inspections and the establishment of "buffer zones" between animal feeding operations and producing fields are the current best practices for environmental assessments. However, a generalized distance guideline and visual inspections may not be enough to account for all environmental risk variables. Here, we report a baseline measurement surveying the background Bacteroidales concentration, as a quantitative fecal contamination indicator, in California's Salinas Valley. We collected a total of 1632 samples from two romaine lettuce commercial fields at the time of harvesting through two seasons in a year. The quantification of Bacteroidales concentration was performed using qPCR, revealing a notably low concentration (0-2.00 copies/cm2) in the commercial fields. To further enhance the applicability of our findings, we developed a user-friendly method for real-world fecal contamination risk assessment that seamlessly integrates with industry practices. Through the generation of heatmaps that visually illustrate varying risk levels across fields, this approach can identify site-specific risks and offer fresh produce stakeholders a more comprehensive understanding of their land. We anticipate this work can encourage the use of Bacteroidales in the fresh produce industry to monitor fecal contamination and prevent future foodborne outbreaks.

A loop-mediated isothermal amplification assay to detect Bacteroidales and assess risk of fecal contamination.

Fecal contamination of fresh produce from human and animal sources is a public health concern due to the risk of foodborne illnesses. The current standard laboratory procedures for microbiological analyses usually require an enrichment step that involves several hours. Molecular techniques such as polymerase chain reaction (PCR) have been used to directly detect pathogens from the samples, however, due to the low quantity of pathogen present and small volumes used for PCR, enrichment is usually required. Additionally, the need for specialized equipment and experienced workers hinders the use of these molecular techniques for field testing. Here, we developed a rapid risk-assessment assay for fecal contamination by targeting Bacteroidales using loop-mediated isothermal amplification (LAMP). The assay allows for naked-eye observation of reactions with as few as ∼8 copies of Bacteroidales per cm2 of the surface in the field. We evaluated this assay with complex field samples as well as on-site field studies. Our on-field studies demonstrated that the Bacteroidales LAMP assay enables us to easily and quickly (<50 min) assess the risk of fecal contamination from animal operations, with a concordance of 85.3% when compared to lab-based qPCR. These results were obtained without expensive equipment (when compared to standard laboratory procedures). These assays could be used to determine site-specific risk and help the decision-making process of fresh produce growers.

The Quest for Immunity: Exploring Human Herpesviruses as Vaccine Vectors.

Herpesviruses are large DNA viruses that have long been used as powerful gene therapy tools. In recent years, the ability of herpesviruses to stimulate both innate and adaptive immune responses has led to their transition to various applications as vaccine vectors. This vaccinology branch is growing at an unprecedented and accelerated rate. To date, human herpesvirus-based vectors have been used in vaccines to combat a variety of infectious agents, including the Ebola virus, foot and mouth disease virus, and human immunodeficiency viruses. Additionally, these vectors are being tested as potential vaccines for cancer-associated antigens. Thanks to advances in recombinant DNA technology, immunology, and genomics, numerous steps in vaccine development have been greatly improved. A better understanding of herpesvirus biology and the interactions between these viruses and the host cells will undoubtedly foster the use of herpesvirus-based vaccine vectors in clinical settings. To overcome the existing drawbacks of these vectors, ongoing research is needed to further advance our knowledge of herpesvirus biology and to develop safer and more effective vaccine vectors. Advanced molecular virology and cell biology techniques must be used to better understand the mechanisms by which herpesviruses manipulate host cells and how viral gene expression is regulated during infection. In this review, we cover the underlying molecular structure of herpesviruses and the strategies used to engineer their genomes to optimize capacity and efficacy as vaccine vectors. Also, we assess the available data on the successful application of herpesvirus-based vaccines for combating diseases such as viral infections and the potential drawbacks and alternative approaches to surmount them.

Water security and health risk assessment of an urban household-level drinking water and sanitation system, Punjab, India.

The water scarcity and deteriorating water quality are major issues of concern to the agrarian state of Punjab, India. The focus of the study is to assess the status of drinking water and sanitation systems of Punjab using an exhaustive dataset of 1575 drinking water samples from 433 sampling locations in 63 urban local bodies of Punjab. Water security index (WSI) indicate that out of 63 urban local bodies, 13 are categorized into good class, 31 fall under fair class, and 19 fall under poor class. The access indicator under sanitation dimension shows that Bathinda region has maximum sewerage network coverage relative to other regions, whereas ca. 50% of the ULBs in Amritsar region do not have sewerage facility. It is clearly depicted that the variation in WSI is mainly attributable to sanitation dimension (10-22.5) as variation in water supply dimension (29-35) is relatively less. Hence, emphasis on indicators and variables of sanitation dimension is required for the improvement of overall WSI. The assessment of qualitative aspects of drinking water and health risk depicts that the drinking water quality of southwest part of the state (i.e. Malwa region) is under good quality class contrary to its poor groundwater quality. Kapurthala district shows high health risk due to the presence of trace metals despite being classified into good class within water security index. The drinking water quality is better and health risks are minimal in regions where drinking water is supplied via treated surface water sources (e.g. Bathinda region). Furthermore, the results of health risk assessment correlate with M[Formula: see text]-Water Quality Index outcome owing to presence of trace metals in groundwater above permissible limits. These results will help in identification of shortcomings in water supply and sanitation infrastructure and its management in urban areas.

On-farm colorimetric detection of Pasteurella multocida, Mannheimia haemolytica, and Histophilus somni in crude bovine nasal samples.

This work modifies a loop-mediated isothermal amplification (LAMP) assay to detect the bovine respiratory disease (BRD) bacterial pathogens Pasteurella multocida, Mannheimia haemolytica, and Histophilus somni in a colorimetric format on a farm. BRD causes a significant health and economic burden worldwide that partially stems from the challenges involved in determining the pathogens causing the disease. Methods such as polymerase chain reaction (PCR) have the potential to identify the causative pathogens but require lab equipment and extensive sample processing making the process lengthy and expensive. To combat this limitation, LAMP allows accurate pathogen detection in unprocessed samples by the naked eye allowing for potentially faster and more precise diagnostics on the farm. The assay developed here offers 66.7-100% analytical sensitivity, and 100% analytical specificity (using contrived samples) while providing 60-100% concordance with PCR results when tested on five steers in a feedlot. The use of a consumer-grade water bath enabled on-farm execution by collecting a nasal swab from cattle and provided a colorimetric result within 60 min. Such an assay holds the potential to provide rapid pen-side diagnostics to cattle producers and veterinarians.

Electron Electric Dipole Moment Searches Using Clock Transitions in Ultracold Molecules.

Permanent electric dipole moments (EDMs) of fundamental particles such as the electron are signatures of parity and time-reversal violation occurring in physics beyond the standard model. EDM measurements probe new physics at energy scales well beyond the reach of present-day colliders. Recent advances in assembling molecules from ultracold atoms have opened up new opportunities for improving the reach of EDM experiments. However, the magnetic field sensitivity of such ultracold molecules means that new measurement techniques are needed before these opportunities can be fully exploited. We present a technique that takes advantage of magnetically insensitive hyperfine clock transitions in polar molecules, offering a way to improve both the precision and accuracy of EDM searches with ultracold assembled molecules.

Genome-wide association mapping identifies markers associated with cane yield components and sucrose traits in the Louisiana sugarcane core collection.

Sugarcane is an economically important crop for both food and biofuel industries. Marker-assisted breeding in sugarcane is becoming a reality with the recent development and deployment of markers linked with disease resistance genes. Large linkage disequilibrium in sugarcane makes genome-wide association studies (GWAS) a better alternative to biparental mapping to identify markers associated with agronomic traits. GWAS was conducted on a Louisiana core collection to identify marker-trait associations (MTA) for 11 cane yield and sucrose traits using single nucleotide polymorphism (SNP) and insertion-deletion (Indel) markers. Significant (P < .05) MTAs were identified for all traits where the top ranked markers explained up to 15% of the total phenotypic variation. High correlations (0.732 to 0.999) were observed between sucrose traits and 56 markers were found consistent across multiple traits. These markers following validation in more diverse populations could be used in marker-assisted selection of clones in sugarcane breeding program in Louisiana and elsewhere.

Discrimination of Proteins Using an Array of Surfactant-Stabilized Gold Nanoparticles.

Protein analysis is a fundamental aspect of biochemical research. Gold nanoparticles are an emerging platform for various biological applications given their high surface area, biocompatibility, and unique optical properties. The colorimetric properties of gold nanoparticles make them ideal for point-of-care diagnostics. Different aspects of gold nanoparticle-protein interactions have been investigated to predict the effect of protein adsorption on colloidal stability, but the role of surfactants is often overlooked, despite their potential to alter both protein and nanoparticle properties. Herein we present a method by which gold nanoparticles can be prepared in various surfactants and used for array-based quantification and identification of proteins. The exchange of surfactant not only changed the zeta potential of those gold nanoparticles but also drastically altered their aggregation response to five different proteins (bovine serum albumin, human serum albumin, immunoglobulin G, lysozyme, and hemoglobin) in a concentration-dependent manner. Finally, we demonstrate that varying surfactant concentration can be used to control assay sensitivity.

A "chemical nose" biosensor for detecting proteins in complex mixtures.

A growing understanding of the fundamental role of proteins in diseases has advanced the development of quantitative protein assays in the medical field. Current techniques for protein analysis include enzyme-linked immunosorbent assays (ELISA), flow cytometry, mass spectrometry, and immunohistochemistry. However, many of these conventional strategies require specialized training, expensive antibodies, or sophisticated equipment, raising assay costs and limiting their application to laboratory analysis. Here, we present the application of a "chemical nose" type colorimetric gold nanoparticle sensor for detection, quantification, and identification of single proteins, protein mixtures, and proteins within the complex environment of human serum. The unique interactions between a mixture of two different gold nanoparticle morphologies (spherical and branched) and six separate proteins (bovine serum albumin, human serum albumin, immunoglobulin G, fibrinogen, lysozyme, and hemoglobin) generated distinguishable protein- and concentration-dependent absorption spectra, even at nanomolar concentrations. Furthermore, we show that this response is sensitive to the relative abundance of different proteins in solution, permitting analysis of protein mixtures. Finally, we demonstrate the ability to distinguish human serum samples with and without a clinically relevant two-fold increase in immunoglobulin G, without the use of expensive reagents or complicated sample processing.

Optimization of Polydiacetylene-Coated Superparamagnetic Magnetite Biosensor for Colorimetric Detection of Biomarkers.

Biosensors for point-of-care testing of critical illnesses are urgently needed, especially in many areas of poor healthcare infrastructure. Polydiacetylene-based sensors are ideal because of their unique colorimetric properties where blue to red color shifts can be observed with the naked eye. In this work, a colorimetric biosensor capable of simple, rapid magnetic separation is optimized, using horse IgG as a model antibody, to obtain higher sensitivity. Composed of a unique combination of polydiacetylene and superparamagnetic iron oxide, the biosensor is fabricated at varying ratios of polydiacetylene to demonstrate optimization of color responsiveness. At increasing polydiacetylene ratios, improved color responsiveness and aqueous dispersion are observed, but the magnetic separation efficiency starts to suffer. The optimal color response is obtained at 90 wt% polydiacetylene. In addition, a 50 times improved lower detection limit of 0.01 mg/mL horse IgG is achieved, a relevant biomarker concentration for diagnosing sepsis. This platform provides a promising colorimetric biosensor for point-of-care use.

U(VI) mitigation via forward osmosis: Elucidation of retention mechanisms and co-ion effects.

Uranium (U) is a chemical and radioactive toxic contaminant affecting many groundwater systems. The focus of this study was to evaluate the suitability of forward osmosis (FO) for uranium rejection from contaminated groundwater under field-relevant conditions. Laboratory experiments with aqueous solution containing uranium were performed with FO membrane to understand the uranium rejection mechanism under varied pH, draw solution concentration, and presence of co-ions. Further, experiments were performed with U-contaminated field groundwater. Results of the hydrogeochemcial modelling using PHREEQC indicated that the rejection mechanism of uranium was highly dependent on aqueous speciation. Uranium rejection was maximum at alkaline pH with ca. 99% rejection due to charge-based interactions between membrane and dominant uranyl complexes. The results of the co-ion study indicated that nitrate and phosphate ions decrease uranium rejection. Whereas, bicarbonates, calcium, and magnesium ions concentrated uranium in feed solution. Further, the uranium adsorption onto the membrane surface primarily depended on pH of the aqueous solution with maximum adsorption at pH 5.5. Our results show that the World Health Organization's drinking water guideline value of 30 µgL-1 for U could be achieved via FO process in field groundwater containing low dissolved solids.

Triangular Osteosynthesis as a Surgical Treatment Option for Unstable Sacral Fracture Fixation with Sacroiliac Dislocation: A Case Report.

Introduction: Sacral fractures, which can occur in young individuals following road traffic accidents or falls from a height, as well as in elderly individuals with osteoporosis after minor trauma, are considered a diverse type of fracture. The incidence of non-osteoporotic sacral fractures is estimated to be 2.1/100,000 people, whereas osteoporotic fractures are estimated to affect 1-5% of elderly individuals at risk. Triangular osteosynthesis is a relatively new fixation technique used as a surgical treatment for unstable sacral fractures. It combines transverse fixation with lumbo-pelvis distraction osteosynthesis, providing stability in different planes. The subcategory of triangular osteosynthesis encompasses spinopelvic fixation, which involves the fusion of transverse sacral alar fracture fixation (such as iliosacral screw/s and sacral plate) and unilateral lumbopelvic fixation from the pedicle of L5 to the ipsilateral posterior ilium. The utilization of this technique provides a mechanically advanced approach for stabilizing unstable sacral alar fractures with vertical shear. Once the pelvic ring injury has been reduced, lumbopelvic fixation can assist in preventing the recurrence of vertical displacement in the unstable hemipelvis. Case Report: The patient, a 29-year-old male, experienced a road traffic accident resulting from a collision involving a motorcycle. As a result of the incident, he suffered from an unstable lateral compression type 1 pelvic ring injury, accompanied by an ipsilateral sacroiliac dislocation and a vertical sacral fracture on the opposite side. Computed tomography imaging revealed a right sacroiliac dissociation, a left sacral fracture classified as AO type B1, as well as fractures in both the superior and inferior pubic rami. The pelvic ring of the patient was subjected to closed reduction and percutaneous fixation, accompanied by minimally invasive spinopelvic fixation. The surgical procedure was performed in a single session, involving the reduction and fixation of the right sacroiliac dissociation, followed by lumbopelvic fixation while in the prone position. After a 1-month follow-up, the patient demonstrated the ability to walk without experiencing pain, and the X-ray revealed a stable spinopelvic and sacroiliac fixation. Conclusion: The utilization of triangular osteosynthesis fixation provides a reliable form of fixation that enables the patient to bear complete weight at an early stage of 6 weeks while also preventing any reduction loss in vertical shear transforaminal sacral fractures.

A Paper-based Loop-Mediated Isothermal Amplification (LAMP) Assay for Highly Pathogenic Avian Influenza.

Avian influenza outbreaks have had significant economic and public health consequences worldwide. Therefore, prompt, reliable, and cost-effective diagnostic devices are crucial for scrutinizing and confining highly pathogenic avian influenza viruses (HPAIVs). Our study introduced and evaluated a novel paper-based loop-mediated isothermal amplification (LAMP) test for diagnosing the H5 subtype of the avian influenza virus (AIV). We meticulously designed and screened LAMP primers targeting the H5-haemagglutinin (H5-HA) gene of AIV and fine-tuned the paper-based detection assay for best performance. The paper-based LAMP assay demonstrated a detection limit of 500 copies per reaction (25 copies/µL). This inexpensive, user-friendly point-of-need diagnostic tool holds great promise, especially in resource-limited settings. It only requires a water bath for incubation and enables visual detection of results without special equipment. Overall, the paper-based LAMP assay provides a promising method for rapidly and reliably detecting the H5 subtype of AIV, contributing to improved surveillance and early intervention strategies.

Strategies for Bovine Respiratory Disease (BRD) Diagnosis and Prognosis: A Comprehensive Overview.

Despite significant advances in vaccination strategies and antibiotic therapy, bovine respiratory disease (BRD) continues to be the leading disease affecting the global cattle industry. The etiology of BRD is complex, often involving multiple microbial agents, which lead to intricate interactions between the host immune system and pathogens during various beef production stages. These interactions present environmental, social, and geographical challenges. Accurate diagnosis is essential for effective disease management. Nevertheless, correct identification of BRD cases remains a daunting challenge for animal health technicians in feedlots. In response to current regulations, there is a growing interest in refining clinical diagnoses of BRD to curb the overuse of antimicrobials. This shift marks a pivotal first step toward establishing a structured diagnostic framework for this disease. This review article provides an update on recent developments and future perspectives in clinical diagnostics and prognostic techniques for BRD, assessing their benefits and limitations. The methods discussed include the evaluation of clinical signs and animal behavior, biomarker analysis, molecular diagnostics, ultrasound imaging, and prognostic modeling. While some techniques show promise as standalone diagnostics, it is likely that a multifaceted approach-leveraging a combination of these methods-will yield the most accurate diagnosis of BRD.

Awareness of Oral and Maxillofacial Surgery as a Career Choice Among Dental Students: An Online Questionnaire Survey.

INTRODUCTION: Most dental students are interested in pursuing a postgraduate degree after completing their Bachelor of Dental Surgery (BDS). Among the various specialties, oral surgery, which bridges the fields of medicine and dentistry, is the most attractive option for many young dental professionals. This study aimed to explore the factors influencing students' choices for postgraduation in oral and maxillofacial surgery (OMFS) and to assess their knowledge, attitudes, and perceptions (KAP) towards this branch of dentistry. MATERIALS AND METHODS: An online survey was conducted with 450 third- and final-year BDS students and interns from various dental colleges. The survey included a self-administered questionnaire designed to collect demographic information and insights into students' motivations and KAP regarding postgraduation in OMFS. The questionnaire was distributed via an internet-based survey program and shared through WhatsApp Messenger accompanied by a consent form. To determine the significance between variables, non-parametric test, such as the chi-square test, was employed. The KAP scores were calculated and represented as mean ± standard deviation (SD), and intergroup comparisons were performed using the two-way independent T-test (for two groups) and two-way analysis of variance (ANOVA) test (for three groups). The level of significance was set at P ≤0.05. RESULTS: The sample comprised 320 (71%) females and 130 (29%) males. A total of 285 (63.33%) respondents wanted to pursue postgraduation after BDS, whereas 130 (28.89%) did not. Long working hours, risk and liability, and lack of skills were the main reasons for not opting for postgraduation in OMFS as a career option. The primary reasons cited for choosing OMFS included better career prospects (75.08%%) and the influence of the mentor (59.65%). The main benefits of choosing postgraduation in OMFS were advanced skills (88.77%), and high earning (85.61%). The mean KAP scores were higher for government institutions in urban areas and for males. CONCLUSION: This study revealed that a substantial number of undergraduate dental students were inclined to specialize in OMFS after graduation. The combination of medicine and dentistry significantly influences the choice of OMFS as a career option.

Porous SnO2 nanosheets for room temperature ammonia sensing in extreme humidity.

Gas sensors based on tin dioxide (SnO2) for the detection of ammonia (NH3) have become commercially available for environmental monitoring due to their reactive qualities when exposed to different gaseous pollutants. Nevertheless, their implementation in the medical field has been hindered by certain inherent drawbacks, such as needing to operate at high temperatures, lack of selectivity, unreliable operation under high-humidity conditions, and a lower detection limit. To counter these issues, this study created 2D nanosheets of SnO2 through an optimized solvothermal method. It was found that tuning the precursor solution's pH to either neutral or 14 led to aggregated or distributed, uniform-size nanosheets with a higher crystallinity, respectively. Remarkably, the SnO2 nanosheet sensor (SNS-14) displayed a much lower response to water molecules and specific reactivity to ammonia even when subjected to reducing and oxidizing agents at 25 °C due to the micropores and chemisorbed oxygen on the nanosheets. Furthermore, the SNS-14 was seen to have the highest sensitivity to ammonia at 100 ppm, with rapid response (8 s) and recovery times (55 s) even at a high relative humidity of 70%. Its theoretical detection limit was recorded to be 64 ppt, better than any of the earlier SnO2-based chemiresistive sensors. Its exceptional sensing abilities were credited to its optimal crystallinity, specific surface area, defects, chemisorbed oxygen, and porous structure. NH3-TPD measurements and computational simulations were employed to understand the ammonia interaction with atomistic details on the SnO2 nanosheet surface. A real time breath sensing experiment was simulated to test the efficacy of the sensor. Reaching this advancement is an achievement in bypassing past boundaries of SnO2-centered sensors, making it feasible to detect ammonia with enhanced precision, discrimination, dependability, and velocity for probable usages in medical diagnostics and ecological surveillance.

Functional Outcomes and Quality of Life After Sacrum Fractures Managed by Either Operative or Conservative Approaches: A Case Series With One-Year Follow-Up.

Background Pelvic fractures caused by high-energy trauma, such as motor vehicle accidents or falls from a considerable height, commonly lead to sacral fractures. Approximately a quarter of sacral fractures are linked to neurological injury, and overlooking these fractures may result in neurological issues such as sexual dysfunction, hindered lower limb functionality, and urinary and rectal difficulties. The main goal of this study is to introduce our patient group who underwent either operative or nonoperative treatment for sacral fractures, with a follow-up period of one year, and assess their functional outcomes. Methodology This is a retrospective review of prospectively collected data from a consecutive series of patients at the Apex Trauma Centre, Sanjay Gandhi Postgraduate Institute of Medical Sciences, Lucknow. A consecutive series of 24 patients (17-55 years old) with sacral fractures treated either operatively or nonoperatively from 2018 to 2023 was studied. A total of 20 patients were available for follow-up questionnaires, and 20 patients participated in a physical examination. Time to final follow-up averaged 27.19 months (range = 12-57 months). The personal data of each patient was collected, including gender, age, comorbidities, concomitant injuries, mechanism of injury, fracture pattern/classification, surgical or nonsurgical treatment, other surgeries, length of surgery, length of hospital stays, adverse events, complications, neurologic and/or motor deficits, bowel and bladder function, and mortality. At a minimum one-year follow-up, the Majeed score, Oswestry Disability Index (ODI) questionnaire, and Gibbon's classification were assessed. Results All fractures were healed. Five patients showed neurological weakness, with three patients having only paresthesia and two patients having lower limb weakness. The mean Majeed score was 75.4, representing a moderate clinical outcome. Final ODI scores averaged 10.6, representing mild disability among patients with sacrum fractures. Overall, 40% of sacrum fractures were associated with sexual dysfunction, with 30% of females and 50% of males reporting this issue. There was no significant difference (p > 0.05) between operated and conservatively managed sacrum fractures concerning ODI scores, neurological deficit, and sexual dysfunction. Conclusions Both male and female patients with traumatic sacrum fractures experienced a significant decrease in their quality of life and sexual function at least 12 months after their surgery. Sacrum fractures are associated with an increased prevalence of sexual dysfunction and bowel/bladder incontinence. Our study findings indicate that patients with sacrum fractures experience similar functional outcomes and incidences of sexual dysfunction irrespective of whether they are managed operatively or conservatively.

A portable, easy-to-use paper-based biosensor for rapid in-field detection of fecal contamination on fresh produce farms.

Laboratory-based nucleic acid amplification tests (NAATs) are highly sensitive and specific, but they require the transportation of samples to centralized testing facilities and have long turnaround times. During the Coronavirus Disease 2019 (COVID-19) pandemic, substantial advancement has been achieved with the development of paper-based point-of-care (POC) NAATs, offering features such as low cost, being easy to use, and providing rapid sample-to-answer times. Although most of the POC NAATs innovations are towards clinical settings, we have developed a portable, paper-based loop-mediated isothermal amplification (LAMP) testing platform for on-farm applications, capable of detecting Bacteroidales as a fecal contamination biomarker. Our integrated platform includes a drop generator, a heating and imaging unit, and paper-based biosensors, providing sensitive results (limit of detection 3 copies of Bacteroidales per cm2) within an hour of sample collection. We evaluated this integrated platform on a commercial lettuce farm with a concordance of 100% when compared to lab-based tests. Our integrated paper-based LAMP testing platform holds great promise as a reliable and convenient tool for on-site NAATs. We expect that this innovation will encourage the fresh produce industry to adopt NAATs as a complementary tool for decision-making in growing and harvesting. We also hope that our work can stimulate further research in the development of on-farm diagnostic tools for other agricultural applications, leading to improved food safety and technology innovation.

Intratumoral localization and activity of 17ß-hydroxysteroid dehydrogenase type 1 in non-small cell lung cancer: a potent prognostic factor.

BACKGROUND: Estrogens were recently demonstrated to be synthesized in non-small cell lung carcinomas (NSCLCs) via aromatase activity and aromatase inhibitor (AI) did suppressed estrogen receptor (ER) positive NSCLC growth. However, other enzymes involved in intratumoral production and metabolism of estrogens, i.e. 17ß-hydroxysteroid dehydrogenases (i.e. 17ßHSD1 and 17ßHSD2) and others have not been studied. Therefore, in this study, we examined the clinical/ biological significance of 17ß-hydroxysteroid dehydrogenases in NSCLCs. METHODOLOGY: Archival materials obtained from 103 NSCLC patients were immunohistochemically evaluated using anti-17ßHSD1 and anti-17ßHSD2 antibodies. The findings of immunohistochemistry were then correlated with intratumoral estrone (E1) and estradiol (E2) concentration, clinicopathological factors and overall survival of the patients. We further employed NSCLC cell lines, A549 and LK87 to study the functional significance of 17ßHSD1, in vitro. RESULTS: A higher 17ßHSD1 immunoreactivity tended to be positively associated with aromatase (p=0.057) and tumor stage (p=0.055) whereas a higher 17ßHSD2 immunoreactivity was positively associated with a squamous cell and adenosquamous cell carcinomas subtypes (p=0.031), tumor stage (p=0.004), T factor of TNM classification (p=0.010), maximum tumor diameter (p=0.002) and tended to be associated with N factor of TMN classification (p=0.065). A higher 17ßHSD1 immunoreactivity was also significantly associated with lower intratumoral E1 concentration (p=0.040) and a higher intratumoral E2/E1 concentration ratio (p=0.028). On the other hand a higher 17ßHSD2 immunoreactivity was significantly associated with higher intratumoral E1 concentration (p=0.035). Results of multivariate regression analysis demonstrated an increased 17ßHSD1 immunoreactivity in tumor cells as an independent negative prognostic factor (HR= 2.83, p=0.007). E1 treatment in 17ßHSD1 positive NSCLC cells, A549 and LK87, resulted in E2 production (p<0.0001) and enhanced cell proliferation, which was abrogated effectively by 17ßHSD1 siRNA knockdown (p<0.0001). In addition, aromatase inhibitor treatment resulted in 17ßHSD1 up regulation in both A549 and LK87 cells. CONCLUSION: Results of our present study suggest that 17ßHSD1 may be considered an important prognostic factor in NSCLC patients and targeting 17ßHSD1 activity may further improve the clinical response in estrogen responsive NSCLC patients.