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INTRODUCTION Radiotherapy is an area of medicine that is little known by patients and that requires a long time for the doctor to be able to explain and resolve the doubts that arise in the first consultation, which may cause anxiety and low adherence to treatment recommendations. A mobile app called Canswer was developed to address this problem in a novel way to help patients learn about radiation oncology in their free time. OBJECTIVE Developed a mobile app that provides oncology radiotherapy information to patients. MATERIAL AND METHODS The patients who attended the consultation for the first time were provided with information and obtained verbal consent, later they were provided with the Canswer application on their mobile device, and at the end, a Likert-type perception survey was applied and the Cancer Treatment Survey (CATS), which analyzed the information needs of patients. Anxiety and depression were analyzed through the Hospital Anxiety and Depression Scale (HADS). RESULTS Of the total number of patients recruited, 17 patients from northeast Mexico with breast cancer agreed that the use of the application would be useful to them and were interested in receiving additional information after their first-time radiotherapy consultation from March to June 2022. Average age of respondents was 45 years. A total of 7 patients (41%) had doubts about radiotherapy after consultation. When analyzing their knowledge, only 1 patient identified that radiotherapy works by DNA damage, the rest had wrong information, 14 patients thought that this treatment works by heating the cells; 2 thought that radiation therapy causes cells to burst. 8 patients (47%) understood how teletherapy works and 5 how brachytherapy works (19%). A question was included about the best skin care during breast radiotherapy in which 76% correctly answered that the skin should be washed daily with water and neutral soap, but 24% answered that hydrating creams should be applied just before stepping into the machine. Regarding the duration of the consultation, a total of 10 patients (60%) thought it was too short. 47% felt the limited time of the consultation did not allow an adequate evaluation. 100% of the respondents were satisfied with the information provided about the type of treatment, its benefits, and adverse effects, however, 47% persisted with doubts at the end of the consultation. 47% of patients (n=8) had some level of anxiety with 29% mild, 11% moderate and 6% severe. Incidence of depression was 24% with 18% having mild form and 6% severe. Regarding the information provided by the doctor during the consultation, 8 patients (47%) would like their doctor to have told them more about anxiety and depression, 7 about sexuality concerns (41%), and 6 (35%) about alternative medicine. 79% of patients wanted to know more about the side effects the treatment might cause after their visit, and 71% reported wanting to know more about how to prevent side effects. 66% also reported that they would like more information about how the treatment feels when it begins. DISCUSSION AND CONCLUSION This analysis revealed that most patients have doubts about radiotherapy, even after the information provided in their first consultation. This may be multifactorial but a key factor may be work overload at radiotherapy centers and a consultation of short duration. This application seems to be an effective, easily accessible, free tool that provides the necessary information, even on topics that are difficult to explain in a standard consultation. Important aspects that the app needs to focus on in future updates include side effects and their management, sexuality, nutrition, depression, and alternative medicine. This application also revealed misconceptions that should be routinely addressed, such as skin care during treatment. Link to app: https://www.canswer.info/?page_id=393 Citation Format: Jose F. Muñoz Lozano, Diana Cristina Pérez Ibave, Estefania Abundis Marquez, Fernando Alcorta Nuñez, Celia B. Gonzalez Alcorta, Carlos Salazar Mejia, Maria Fernanda Noriega, Omar Zayas Villanueva, Victor Oyervides Juarez, Larisa M. Renteria Garcia, Adelina Alcorta Garza, Juan Francisco Gonzalez Guerrero, Valeria Luna, David Hernandez, Rafael Piñeiro Retif, Oscar Vidal Gutiérrez. EXPLORATORY ANALYSIS OF A MOBILE APP THAT ADDRESSES RADIOTHERAPY INFORMATIONAL NEEDS FOR BREAST CANCER PATIENTS [abstract]. In: Proceedings of the 2022 San Antonio Breast Cancer Symposium; 2022 Dec 6-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2023;83(5 Suppl):Abstract nr P1-08-03.

Hereditary cancer syndromes (HCS) are genetic diseases with an increased risk of developing cancer. This research describes the implementation of a cancer prevention model, genetic counseling, and germline variants testing in an oncologic center in Mexico. A total of 315 patients received genetic counseling, genetic testing was offered, and 205 individuals were tested for HCS. In 6 years, 131 (63.90%) probands and 74 (36.09%) relatives were tested. Among the probands, we found that 85 (63.9%) had at least one germline variant. We identified founder mutations in BRCA1 and a novel variant in APC that led to the creation of an in-house detection process for the whole family. The most frequent syndrome was hereditary breast and ovarian cancer syndrome (HBOC) (41 cases with BRCA1 germline variants in most of the cases), followed by eight cases of hereditary non-polyposic cancer syndrome (HNPCC or Lynch syndrome) (with MLH1 as the primarily responsible gene), and other high cancer risk syndromes. Genetic counseling in HCS is still a global challenge. Multigene panels are an essential tool to detect the variants frequency. Our program has a high detection rate of probands with HCS and pathogenic variants (40%), compared with other reports that detect 10% in other populations.

Introduction A minority of women with breast cancer in Mexico are diagnosed through a screening program with a low rate of national coverage of about 20%, which translates into late diagnosis and worse outcomes. The detection of high-risk groups through easy-access interventions is essential to increase the screening rate. OBJECTIVE To identify and analyze patients at high risk of breast cancer, through a web app specially designed for this purpose. MATERIAL AND METHODS The web app stratified respondents according to breast cancer risk into 4 categories: very high risk (symptomatic), high lifetime risk, average risk (usual screening recommendations), or low risk. The app was programmed to guide patients to a risk-based information page or urge them to seek medical advice if indicated (https://cuccuanl.com/tamiz_mama/). The web app also provided the contact information of our center, or an appointment could also be scheduled within the app. Distribution was made via social media. An upgrade of the app was launched, which expanded the functionality with the inclusion of the Gail model to identify women with a high lifetime risk for developing breast cancer and the option for a simple interpretation of mammographic results. In version 2.0, the patients were divided into 2 populations, the first one which already had a breast imaging test but wished to know what the BIRADS score obtained meant and the other group were those who answered a simple survey similar to the one available on version 1.0, with added questions to calculate the Gail model risk. RESULTS The web app was originally released in October 2021 with a total number of 1,012 women answering the survey. The update was released in June 2022 with 406 new subjects. 281 patients wished for a risk calculation with no prior breast imaging tests and 124 patients already had a breast image diagnosis test that wanted a simple interpretation. Table1 depicts the answers given between both versions. Among the group of patients that had a mammography but had doubts about the result 12.9% (n=16) reported a BIRADS 0, 12.1% (n=15) BIRADS 1, 46% (n=57) BIRADS 2, 14.5% (n=18) BIRADS 3, and 14.4% (n=18) BIRADS 4 or 5. To date 41 patients have booked an appointment at the cancer prevention clinic directly within the webapp for further evaluation. DISCUSSION AND CONCLUSION The changes in the criteria of high risk did not translate into changes in the proportion of patients considered at high lifetime risk of breast cancer, but these changes made might be more specific for subsequent screening and chemoprevention strategies. The app persisted with a high proportion of symptomatic patients. The results of integration of the simple mammography interpretation were surprising, with a high proportion of subjects with indication for a biopsy founded. In the Mexican population there is a significant gap between the screening mammography and the first consultation with a specialist, averaging 113 days from an abnormal screening test to a diagnosis of breast cancer (1), which delays treatment initiation resulting on a worst outcome. These kinds of apps may empower patients to seek earlier consultation if warranted or educate patients on their personal risk for developing cancer so they may have a closer adherence to early detection programs. These results and subsequent updates might help evolve the app into something more akin to a digital navigator for patients. Bibliography 1)Unger-Saldaña K, Cedano Guadiamos M. Delays to diagnosis and barriers to care for breast cancer in Mexico and Peru: a cross sectional study. The Lancet Global Health. 2020 Apr 1;8:S16. Table 1 Results stratified by risk groups between version 1.0 and 2.0 Citation Format: Jose F. Muñoz Lozano, Omar Zayas Villanueva, Estefania Abundis Marquez, Fernando Alcorta Nuñez, Maria Fernanda Noriega, Carlos Salazar Mejia, Celia B. Gonzalez Alcorta, Diana Cristina Pérez Ibave, Victor Oyervides Juarez, Larisa M. Renteria Garcia, Adelina Alcorta Garza, Juan Francisco Gonzalez Guerrero, David Hernandez, Rafael Piñeiro Retif, Oscar Vidal Gutiérrez. Analysis of an update to a novel breast cancer screening web-app [abstract]. In: Proceedings of the 2022 San Antonio Breast Cancer Symposium; 2022 Dec 6-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2023;83(5 Suppl):Abstract nr P5-04-09.

Breast cancer (BC) has one of the highest incidences and mortality worldwide. Single nucleotide polymorphisms (SNPs) in

1567 Background: A minority of women with breast cancer in Mexico are being diagnosed through a screening program, which translates into late diagnosis and a worse prognosis. It is imperative to develop easy access and low-cost interventions to increase the screening rate. Methods: We developed a web app tailored to guide patients to seek medical consultation if they were at high risk for breast cancer or standard screening recommendations based on the predicted risk for age (https://cuccuanl.com/calcula-tu-riesgo-de-cancer-de-mama/). The web app consisted of an 8-question survey designed using object-based programming with HTML5, the additional logic was programmed on JavaScript. The program logic automatically guided patients to medical consultation if they were at high risk for breast cancer, or standard screening recommendations based on their predicted risk for age. On each confirmation page, the contact information of the hospital appeared, also, consultation at the clinic could be scheduled within the web app. The web app was distributed via social media on International Breast Cancer Awareness Day. Results: A total of 1,012 persons answered the survey after a follow-up period of two weeks. The median age of respondents was 34 years. Among participants, 10.8% were considered at very high risk for breast cancer due to symptoms, 22% were classified as high risk based on family history or more than 5 years of contraceptive use, and 19% were considered as average-risk population for whom age-based screening tests were recommended. The remaining 48% of participants were considered at low risk for breast cancer development and were directed to educational information about breast cancer awareness. Among all persons that answered our survey, 21 requested a specialized medical appointment within the web app. Conclusions: This pilot study showed a high response of Mexican population seeking breast cancer risk evaluation, 11% of women that responded to our survey had symptoms highly suggestive of breast cancer. The use of web apps can result in mass diffusion which will help reach people with less medical access, a common scenario in many developing countries. Further analysis should be made to measure the real impact on breast cancer diagnosis and oncological outcomes. This study shows how effective social media is as a means of diffusion on health topics. [Table: see text]

1548 Background: Lung cancer screening continues to be an area of great opportunity in public health, especially in developing countries. In Mexico, about 10,000 new cases of lung cancer are detected annually, of which less than 5% are diagnosed in early stages. There are no national programs for timely detection of lung cancer in our country, so it is essential to seek accessible measures to prioritize resources for high-risk people. Methods: We developed a web app that consisted of a short survey to stratify patients according to their risk of lung cancer (https://cuccuanl.com/tamiz_pulmon/). The questions were based on Nelson's criteria to categorize high-risk subjects who are deemed candidates for screening. The program contained automated logic programmed using JavaScript to guide people to the low or high-risk page if they met standard risk criteria in those age 50 and older. The high-risk page alerted people of their risk, displaying general information about lung cancer as well as contact information to make an appointment at our cancer center. An appointment could be also scheduled within the app if the person so wished. The web app was launched and distributed through social media. Results: After a period of 2 months, 939 people completed the survey. The median age of the responders was 40 years, and 61% were men. Of the total, 185 participants were 50 years of age or older. 268 people (29% of the total) were sent to the high-risk page, including persons under 50 years of age with symptoms highly suggestive of lung cancer. According to their smoking status, 80% of the subjects reported active smoking, while 9% reported heavy smoking, considered as more than 20 pack-years. Among all the people evaluated, 44 high-risk subjects scheduled a specialized medical appointment within the web app. Conclusions: This pilot study showed a high response of Mexican population seeking lung cancer risk evaluation, especially among the persons who smoke regularly. The use of web apps can result in mass diffusion which will help reach people with less medical access, a common scenario in many developing countries. Further analysis should be made to measure the real impact on lung cancer diagnosis and oncological outcomes. Our study shows how effective social media is as a means of diffusion on health topics. [Table: see text][Table: see text]

Human papillomavirus (HPV) DNA integration is a crucial event in cervical carcinogenesis. However, scarce studies have focused on studying HPV integration (HPVint) in early-stage cervical lesions. Using HPV capture followed by sequencing, we investigated HPVint in pre-tumor cervical lesions. Employing a novel pipeline, we analyzed reads containing direct evidence of the integration breakpoint. We observed multiple HPV infections in most of the samples (92%) with a median integration rate of 0.06% relative to HPV mapped reads corresponding to two or more sequence breakages. Unlike cancer studies, most integrations events were unique (supported by one read), consistent with the lack of clonal selection. Congruent to other studies, we found that breakpoints could occur, practically, in any part of the viral genome. We noted that L1 had a higher frequency of rupture integration (25%). Based on host genome integration frequencies, we found previously reported integration sites in cancer for genes like FHIT, CSMD1, and LRP1B and putatively many new ones such as those exemplified in CSMD3, ROBO2, and SETD3. Similar host integrations regions and genes were observed in diverse HPV types within many genes and even equivalent integration positions in different samples and HPV types. Interestingly, we noted an enrichment of integrations in most centromeres, suggesting a possible mechanism where HPV exploits this structural machinery to facilitate integration. Supported by previous findings, overall, our analysis provides novel information and insights about HPVint.

Familial adenomatous polyposis (FAP) is an autosomal-dominant condition characterized by the presence of multiple colorectal adenomas, caused by germline variants in the adenomatous polyposis coli (APC) gene. More than 300 germline variants have been characterized. The detection of novel variants is important to understand the mechanisms of pathophysiology. We identified a novel pathogenic germline variant using next-generation sequencing (NGS) in a proband patient. The variant is a complex rearrangement (c.422+1123_532-577 del ins 423-1933_423-1687 inv) that generates a complete deletion of exon 5 of the APC gene. To study the variant in other family members, we designed an endpoint PCR method followed by Sanger sequencing. The variant was identified in the proband patient’s mother, one daughter, her brother, two cousins, a niece, and a second nephew. In patients where the variant was identified, we found atypical clinical symptoms, including mandibular, ovarian, breast, pancreatic, and gastric cancer. Genetic counseling and cancer prevention strategies were provided for the family. According to the American College of Medical Genetics (ACMG) guidelines, this novel variant is considered a PVS1 variant (very strong evidence of pathogenicity), and it can be useful in association with clinical data for early surveillance and suitable treatment.

The southern house mosquito, Culex quinquefasciatus Say (Diptera: Culicidae), transmits pathogens and is one of the most important vectors of human disease. The carboxylesterase mosquito genes encoded esterase enzyme involved in the mechanism that provides resistance to insecticides. We amplified by RTPCR four alternative transcripts from the esterase gene not reported previously (A, B, C, and D) from Cx. quinquefasciatus larvae from Monterrey, N.L., northeastern Mexico. Through cDNA sequences, we predicted amino acid sequences used to model the three-dimensional structure of protein isoforms. We determined dN and dS ratios to identify evolutionary forces which lead the family. PCR products were obtained from 5A esterases (1839, 1898, 1894, and 1953 bp). The translation sequence of the PCR products showed polypeptides of 612, 473, 282, and 282 amino acids in length for isoforms A, B, C, and D, respectively. Three exons and two introns were obtained in the analyzed sequences. The study sequences were compared with those previously published showing 98.7, 98.5, 91.5, and 91.5% amino acid similarity for isoforms A, B, C, and D, respectively. The threedimensional models showed that only ORF-D had the three amino acids corresponding to the catalytic triad, while ORF-A and ORF-C lacked one or two of the three amino acids. Esterase genes are under purifying of selection; a clue the members are functional. The processes might be useful in monitoring evolutionary dynamics of known carboxylesterase genes and identifying new carboxylesterase splicing.

Aspergillus is an opportunistic fungus present in humid environments, whose natural environment is in soil, hay and compost. It is a frequent contaminant in the clinical laboratory. Because of this, the fungus is often inhaled, affecting those with an underlying pulmonary disease or immune deficiency. Fungal genitourinary tract infections are relatively common. A rare Aspergillus spp cervical infection diagnosed via liquid-based cytology is presented in the current study. The 57-year-old woman attended her annual check-up without any relevant medical history. The result of a gynecological examination by Papanicolaou smear was normal and routine liquid-based cytology was performed. The specimen exhibited fungal organisms characterized by septate hyphae branching at acute angles, most consistent with the Aspergillus species. Subsequent cytology demonstrated the same results. Antifungal treatment was initiated and a second post-treatment smear only exhibited atrophy. The cytomorphological features of Aspergillus spp. are discussed in the current study and a brief review of the few reported cases of a primary cervical infection in the literature is provided. In addition, the liquid-based cytology was established as a tool to diagnose the rare Aspergillus infection.

Olfactomedin-like (OLFML) proteins are members of the olfactomedin domain-containing secreted glycoprotein (OLF) family. OLFML2A and OLFML2B are representative molecules of these glycoproteins. Olfactomedins are critical for the development and functional organization of the nervous system and retina, which is a highly conserved structure in vertebrates, having almost identical anatomical and physiological characteristics in multiple taxa. Spotted gar, a member of the Lepisosteidae family, is a freshwater fish that inhabits rivers, bayous, swamps, and brackish waters. Recently, the complete genome has been sequenced, providing a unique bridge between fish medical models to human biology, making it an excellent animal model. This study was aimed to understanding the evolution OLFML2A and OLFML2B in the retina of spotted gar through looking for the expression of these genes. Spotted gar retina was analyzed with hematoxylin-eosin staining assays to provide an overall view of the retina structure and an immunofluorescence assay to identify OLFML2A and OLFML2B protein expression. A phylogenetic tree was created using the neighbor-joining method. Forces that direct the evolution of the fish genes were tested. Spotted gar retina, as in other vertebrates, is made of several layers. OLFML2A and OLFML2B proteins were detected in the rod and cone photoreceptor layer (PRL), outer nuclear layer (ONL), and inner nuclear layer (INL). Phylogenetic tree analysis confirms the orthology within the OLFML2A gene. Purifying selection is the evolutionary force that directs the OLFML2A genes. OLFML2A genes have a well-conserved function over time and species.

In humans, the polygenic growth hormone (GH) locus is located on chromosome 17 and contributes with three types of proteins: pituitary GH which consists of at least two isoforms one of 22 kDa and the other of 20 kDa, placental GH, which also exhibits isoforms, and chorionic somatomammotropin hormone (CSH). While pituitary GH results from the expression of the GH-1 (GH-N) gene, placental GH is produced by the expression of the GH-2 (GH-V) gene and CSH is contributed by expression of the CSH-1 and CSH-2 genes. The location where GH-1 is expressed is the anterior pituitary and the rest of the genes in the locus are expressed in placenta. On the other hand, expression and synthesis of GH in extra-pituitary tissues, including the eye, has been recently described. However, the physiological role of GH in the eye has not yet been elucidated, although a possible neuroprotective role has been hypothesized. Thus, we analyzed GH-1, GH-2, CSH1/2, Pit-1, GHR, GHRH, GHRHR, SST, SSTR1, SSTR2, SSTR3, SSTR4, and SSTR5 to elucidate the expression and regulation of the GH locus in the human eye. Through qPCR analysis, we only found evidence of GH-1 expression in retina, choroid and trabecular meshwork; its transcript turned out to be the same as pituitary GH mRNA found in major species, and no splicing variants were detected. PIT1 was absent in all the ocular tissues implying an independent GH-1 expression mechanism. We found evidence of GHR in the cornea, choroid coat and retina. These results suggest autocrine and/or paracrine regulation, possibly exerted by GHRH and SSTs (since their mRNAs and receptors were found predominantly in retinal, choroidal and corneal tissues) since expression of both molecules was detected in different ocular tissues, as well as in the same tissues where GH-1 expression was confirmed. Our results add solid evidence about the existence of a regulatory local system for GH expression and release in the human eye.

Prostate-specific antigen (PSA) is a serine protease produced by epithelial prostatic cells and its main function is to liquefy seminal coagulum. Currently, PSA is a biomarker for the diagnosis and screening of prostate cancer and it was the first cancer biomarker approved by the FDA. The quantity and serum isoforms of male PSA, allows distinguishing between carcinoma and benign inflammatory disease of the prostate. Initially, it was thought that PSA was produced only by the prostate, and thus, a protein that was expressed exclusively in men. However, several authors report that PSA is a protein that is expressed by multiple non-prostatic tissues not only in men but also in women. Some authors also report that in women, the expression of this protein is highly related to breast and colon cancer and therefore can act as a possible biomarker for early detection, diagnosis and prognosis of these cancers in women. In this review, we will focus on the characteristics of the PSA at a molecular level, its current clinical implications, the expression of this protein in non-prostatic tissues, and its relationship with cancer, especially in women.

The human growth hormone (GH) locus is comprised by two GH (GH1 and GH2) genes and three chorionic somatomammotropin (CSH1, CSH2 and CSH-L) genes. While GH1 is expressed in the pituitary gland, the rest are expressed in the placenta. However, GH1 is also expressed in several extrapituitary tissues, including the eye. So to understand the role of this hormone in the eye we used the baboon (Papio hamadryas), that like humans has a multigenic GH locus; we set up to investigate the expression and regulation of GH locus in adult and fetal baboon ocular tissues. We searched in baboon ocular tissues the expression of GH1, GH2, CSH1/2, Pit1 (pituitary transcription factor 1), GHR (growth hormone receptor), GHRH (growth hormone releasing hormone), GHRHR (growth hormone releasing hormone receptor), SST (somatostatin), SSTR1 (somatostatin receptor 1), SSTR2 (somatostatin receptor 2), SSTR3 (somatostatin receptor 3), SSTR4 (somatostatin receptor 4), and SSTR5 (somatostatin receptor 5) mRNA transcripts and derived proteins, by qPCR and immunofluorescence assays, respectively. The transcripts found were characterized by cDNA cloning and sequencing, having found only the one belonging to GH1 gene, mainly in the retina/choroid tissues. Through immunofluorescence assays the presence of GH1 and GHR proteins was confirmed in several retinal cell layers. Among the possible neuroendocrine regulators that may control local GH1 expression are GHRH and SST, since their mRNAs and proteins were found mainly in the retina/choroid tissues, as well as their corresponding receptors (GHRH and SSTR1-SSTR5). None of the ocular tissues express Pit1, so gene expression of GH1 in baboon eye could be independent of Pit1. We conclude that to understand the regulation of GH in the human eye, the baboon offers a very good experimental model.

We evaluated the participation of oxidative stress in the negative regulation of hepatitis C virus (HCV)-RNA induced by acetylsalicylic acid (ASA). We used the HCV subgenomic replicon cell system that stably expresses HCV-nonstructural proteins (Huh7 HCV replicon cells) and the parental cell line. Cells were exposed to 4 mM ASA at different times (12–72 h), and pyrrolidine dithiocarbamate (PDTC) was used as an antioxidant control. Reactive oxygen species (ROS) production, oxidized protein levels, cytosolic superoxide dismutase (Cu/Zn-SOD), and glutathione peroxidase (GPx) activity were measured to evaluate oxidative stress. In addition, viral RNA and prostaglandin (PGE2) levels were determined. We observed that ASA treatment decreased ROS production and oxidized protein levels in a time-dependent fashion in both parental and HCV replicon cells with a greater extent in the latter. Similar results were found with PDTC exposure. Average GPx activity was decreased, whereas a striking increase was observed in average cytosolic SOD activity at 48 and 72 h in both cells exposed to ASA, compared with untreated cells. HCV replicon cells showed higher levels of Cu/Zn-SOD expression (mRNA and protein) with ASA treatment (48 and 72 h), whereas NS5A protein levels showed decreased expression. In addition, we found that inhibition of SOD1 expression reversed the effect of ASA. Interestingly, PDTC downregulated HCV-RNA expression (55%) and PGE2 (60%) levels, imitating ASA exposure. These results suggest that ASA treatment could reduce cellular oxidative stress markers and modify Cu/Zn-SOD expression, a phenomenon that may contribute to the mechanisms involved in HCV downregulation.

Background: Our aim was to develop a quantitative real-time polymerase chain reaction (qPCR) assay for the quantitation of hepatitis C virus (HCV) RNA samples from patients infected with different HCV genotypes. Methods: A standard curve was generated by amplification of serial dilutions of HCV-plasmid (pFKI389-NS3-3′) harboring genotype 1b HCV-subgenomic replicon. Samples from 15 HCV-infected patients (genotypes 1, 2, and 3) were analyzed to quantify HCV-RNA by qPCR with primers and probes specific for the 5′-UTR viral region. Results: The HCV qPCR assay had a sensitivity of 100 copies/reaction with a dynamic range of detection between 102–20 × 106 HCV copies. The assay was highly reproducible with a low coefficient of variation. We observed that the HCV genotypes included could be identified by our method. Conclusions: Our results showed that this modified qPCR assay provides a valid platform for quantifying HCV-RNA, combining good analytical sensitivity with a wide dynamic range and high reproducibility. * qPCR : quantitative real-time polymerase chain reaction; HCV : hepatitis C virus; PEG-IFN-α : pegylated interferon alpha; SVR : sustained virological response; IU : international units; ART : Abbott RealTime HCV assay; CAP/CTM : Roche Cobas AmpliPrep/Cobas TaqMan HCV assay; RT-PCR : reverse transcription polymerase chain reaction; DEPC : diethylpyrocarbonate; Ct : threshold cycle; SD : standard deviations; CV : coefficients of variation

Background Olfactomedin-like is a family of polyfunctional polymeric glycoproteins. This family has at least four members. One member of this family is OLFML3, which is preferentially expressed in placenta but is also detected in other adult tissues including the liver and heart. However, its orthologous rat gene is expressed in the iris, sclera, trabecular meshwork, retina, and optic nerve. Methods OLFML3 messenger amplification was performed by RT-PCR from human and baboon ocular tissues. The products were cloned and sequenced. Results We report OLFML3 expression in human and baboon eye. The full coding DNA sequence has 1221 bp, from which an open reading frame of 406 amino acid was obtained. The baboon OLFML3 gene nucleotidic sequence has 98% and amino acidic 99% similarity with humans. Conclusions OLFML3 gene expression in human and baboon ocular tissues and its high similarity make the baboon a powerful model to deduce the physiological and/or metabolic function of this protein in the eye.

Background The olfactomedin-like domain (OLFML) is present in at least four families of proteins, including OLFML2A and OLFML2B, which are expressed in adult rat retina cells. However, no expression of their orthologous has ever been reported in human and baboon. Objective The aim of this study was to investigate the expression of OLFML2A and OLFML2B in ocular tissues of baboons (Papio hamadryas) and humans, as a key to elucidate OLFML function in eye physiology. Methods OLFML2A and OLFML2B cDNA detection in ocular tissues of these species was performed by RT-PCR. The amplicons were cloned and sequenced, phylogenetically analyzed and their proteins products were confirmed by immunofluorescence assays. Results OLFML2A and OLFML2B transcripts were found in human cornea, lens and retina and in baboon cornea, lens, iris and retina. The baboon OLFML2A and OLFML2B ORF sequences have 96% similarity with their human’s orthologous. OLFML2A and OLFML2B evolution fits the hypothesis of purifying selection. Phylogenetic analysis shows clear orthology in OLFML2A genes, while OLFML2B orthology is not clear. Conclusions Expression of OLFML2A and OLFML2B in human and baboon ocular tissues, including their high similarity, make the baboon a powerful model to deduce the physiological and/or metabolic function of these proteins in the eye.

The gene for pituitary growth hormone (GH-N) in man belongs to a multigene locus located at chromosome 17q24.2, which also harbors four additional genes: one for a placental variant of GH-N (named GH-V) and three of chorionic somatommamotropin (CSH) type. Their tandem arrangement from 5' to 3' is: GH-N, CSH-L, CSH-1, GH-V and CSH-2. GH-N is mainly expressed in the pituitary from birth throughout life, while the remaining genes are expressed in the placenta of pregnant women. Pituitary somatotrophs secrete GH into the bloodstream to act at receptor sites in most tissues. GH participates in the regulation of several complex physiological processes, including growth and metabolism. Recently, the presence of GH has been described in several extrapituitary sites, such as neural, ocular, reproductive, immune, cardiovascular, muscular, dermal and skeletal tissues. It has been proposed that GH has an autocrine action in these tissues. While the body of evidence for its presence is constantly growing, research of its possible function and implications lag behind. In this review we highlight the evidence of extrapituitary synthesis of GH in humans.