Thermostable in vitro transcription-translation compatible with microfluidic droplets.

BACKGROUND: In vitro expression involves the utilization of the cellular transcription and translation machinery in an acellular context to produce one or more proteins of interest and has found widespread application in synthetic biology and in pharmaceutical biomanufacturing. Most in vitro expression systems available are active at moderate temperatures, but to screen large libraries of natural or artificial genetic diversity for highly thermostable enzymes or enzyme variants, it is instrumental to enable protein synthesis at high temperatures. OBJECTIVES: Develop an in vitro expression system operating at high temperatures compatible with enzymatic assays and with technologies that enable ultrahigh-throughput protein expression in reduced volumes, such as microfluidic water-in-oil (w/o) droplets. RESULTS: We produced cell-free extracts from Thermus thermophilus for in vitro translation including thermostable enzymatic cascades for energy regeneration and a moderately thermostable RNA polymerase for transcription, which ultimately limited the temperature of protein synthesis. The yield was comparable or superior to other thermostable in vitro expression systems, while the preparation procedure is much simpler and can be suited to different Thermus thermophilus strains. Furthermore, these extracts have enabled in vitro expression in microfluidic droplets at high temperatures for the first time. CONCLUSIONS: Cell-free extracts from Thermus thermophilus represent a simpler alternative to heavily optimized or pure component thermostable in vitro expression systems. Moreover, due to their compatibility with droplet microfluidics and enzyme assays at high temperatures, the reported system represents a convenient gateway for enzyme screening at higher temperatures with ultrahigh-throughput.

Wound closure after knee and hip replacement (TKA and THA): Survey results on the clinical practice in Spain. / Cierre de herida en cirugía protésica de rodilla y cadera (PTR y PTC): Resultados de una encuesta sobre la práctica clínica en España.

BACKGROUND AND OBJECTIVE: In orthopedic surgery, it is clear that an optimal standardized closure technique has not yet been developed. Locally, there are no objective data describing the standard surgical practice in wound closure. The aim of this study is to analyze the clinical practice of surgical wound closure in orthopedic surgery by means of a survey of a representative local sample and thus obtain information on the context of closure in Spain. METHOD: an ad hoc group of specialists in orthopedic surgery and traumatology was formed. The group of experts, after analyzing the literature, developed a questionnaire of 32 closed multiple-choice questions divided into the following blocks: hemostasis, surgical wound closure (deep, superficial, and cutaneous), and dressings. RESULTS: A total of 471 surgeons responded to the survey completely and with sufficient information to perform the descriptive analysis. 79% believe that ATX can influence the decrease in surgical site infection rate. 96% believe that the type of deep closure at the level of the arthrotomy could influence outcomes and complications after hip and/or knee replacements. 85% believe that the type of shallow closure at the subcutaneous level may influence outcomes and complications after hip and/or knee replacement. 64% of surgeons use single-use incisional negative pressure therapy for the treatment of surgical wound complications (seroma, prolonged drainage, dehiscence). CONCLUSIONS: There is a high level of variability in wound closure in our setting and a low level of training on the subject. The authors recommend that the different scientific societies invest resources to improve training in this field and reduce the percentage of surgeons who are considered inadequately trained, as well as adapting closure techniques to those considered gold standard according to the evidence.

[Translated article] Wound closure after knee and hip replacement (TKA and THA): Survey results on the clinical practice in Spain.

BACKGROUND AND OBJECTIVE: In orthopaedic surgery, it is clear that an optimal standardised closure technique has not yet been developed. Locally, there are no objective data describing the standard surgical practice in wound closure. The aim of this study is to analyse the clinical practice of surgical wound closure in orthopaedic surgery by means of a survey of a representative local sample and thus obtain information on the context of closure in Spain. MATERIAL AND METHOD: An ad hoc group of specialists in orthopaedic surgery and traumatology was formed. The group of experts, after analyzing the literature, developed a questionnaire of 32 closed multiple-choice questions divided into the following blocks: hemostasis, surgical wound closure (deep, superficial, and cutaneous), and dressings. RESULTS: A total of 471 surgeons responded to the survey completely and with sufficient information to perform the descriptive analysis. 79% believe that ATX can influence the decrease in surgical site infection rate. 96% believe that the type of deep closure at the level of the arthrotomy could influence outcomes and complications after hip and/or knee replacements. 85% believe that the type of shallow closure at the subcutaneous level may influence outcomes and complications after hip and/or knee replacement. 64% of surgeons use single-use incisional negative pressure therapy for the treatment of surgical wound complications (seroma, prolonged drainage, dehiscence). CONCLUSIONS: There is a high level of variability in wound closure in our setting and a low level of training on the subject. The authors recommend that the different scientific societies invest resources to improve training in this field and reduce the percentage of surgeons who are considered inadequately trained, as well as adapting closure techniques to those considered gold standard according to the evidence.

Reporting serious incidents in medical devices used in intraocular surgery: Proposing ideas.

The entry into force of the regulation on medical devices obliges clinicians to identify and report to the Health Authorities possible serious incidents arising from their use. In view of the doubts that may arise as to whether or not it may be considered a serious incident, a working group, set up by members of the Spanish Society of Retina and Vitreo and the cluster of ophthalmology and vision sciences (Cluster4Eye) have prepared a document that aims to guide ophthalmologists about some of the incidents that, in the experience of the work team, are not common or can cause serious damage to the patient's function.

Quantification of Microvascular Lesions in the Central Retinal Field: Could It Predict the Severity of Diabetic Retinopathy?

Diabetic retinopathy (DR) is a neurodegenerative disease characterized by the presence of microcirculatory lesions. Among them, microaneurysms (MAs) are the first observable hallmark of early ophthalmological changes. The present work aims to study whether the quantification of MAs, hemorrhages (Hmas) and hard exudates (HEs) in the central retinal field could have a predictive value on DR severity. These retinal lesions were quantified in a single field NM-1 of 160 retinographies of diabetic patients from the IOBA's reading center. Samples included different disease severity levels and excluded proliferating forms: no DR (n = 30), mild non-proliferative (n = 30), moderate (n = 50) and severe (n = 50). Quantification of MAs, Hmas, and HEs revealed an increasing trend as DR severity progresses. Differences between severity levels were statistically significant, suggesting that the analysis of the central field provides valuable information on severity level and could be used as a clinical tool to assess DR grading in the eyecare routine. Even though further validation is needed, counting microvascular lesions in a single retinal field can be proposed as a rapid screening system to classify DR patients with different stages of severity according to the international classification.

[Translated article] Overview of systematic reviews of risk factors for prosthetic joint infection.

BACKGROUND: Prosthetic joint infection is one of the most serious complications in orthopedics. Prognostic systematic reviews (SRs) detecting and assessing factors related to prosthetic joint infection, allow better prediction of risk and implementation of preventive measures. Although prognostic SRs are increasingly frequent, their methodological field presents some knowledge gaps. PURPOSE: To carry out an overview of SR assessing risk factors for prosthetic joint infection, describing and synthesizing their evidence. Secondarily, to assess the risk of bias and methodological quality. MATERIAL AND METHODS: We conducted a bibliographic search in 4 databases (May 2021) to identify prognostic SR evaluating any risk factor for prosthetic joint infection. We evaluated risk of bias with the ROBIS tool, and methodological quality with a modified AMSTAR-2 tool. We computed the overlap degree study between included SR. RESULTS: Twenty-three SRs were included, studying 15 factors for prosthetic joint infection, of which, 13 had significant association. The most frequently studied risk factors were obesity, intra-articular corticosteroids, smoking and uncontrolled diabetes. Overlapping between SR was high for obesity and very high for intra-articular corticoid injection, smoking and uncontrolled diabetes. Risk of bias was considered low in 8 SRs (34.7%). The modified AMSTAR-2 tool showed important methodological gaps. CONCLUSIONS: Identification of procedural-modifiable factors, such as intra-articular corticoids use, can give patients better results. Overlapping between SR was very high, meaning that some SRs are redundant. The evidence on risk factors for prosthetic joint infection is weak due to high risk of bias and limited methodological quality.

Mercury intoxication and ophthalmic involvement: An update review.

Human intoxication after mercury exposure is a rare condition that can cause severe damage to the central nervous, respiratory, cardiovascular, renal, gastrointestinal, skin, and visual systems and represents a major public health concern. Ophthalmic involvement includes impaired function of the extraocular muscles and the eyelids, as well as structural changes in the ocular surface, lens, retina, and optic nerve causing a potential irreversible damage to the visual system. Although, there are many pathways for poisoning depending on the mercury form, it has been suggested that tissue distribution does not differ in experimental animals when administered as mercury vapor, organic mercury, or inorganic mercury. Additionally, visual function alterations regarding central visual acuity, color discrimination, contrast sensitivity, visual field and electroretinogram responses have also been described widely. Nevertheless, there is still controversy about whether visual manifestations occur secondary to brain damage or as a direct affectation, and which ocular structure is primarily affected. Despite the use of some imaging techniques such as in vivo confocal microscopy of the cornea, optical coherence tomography (OCT) of the retina and optic nerve, and functional tests such as electroretinography has helped to solve in part this debate, further studies incorporating other imaging modalities such as autofluorescence, OCT angiography or adaptive optics retinal imaging are needed. This review aims to summarize the published structural and functional alterations found in the visual system of patients suffering from mercury intoxication.

Overview of systematic reviews of risk factors for prosthetic joint infection. / Revisión de revisiones sistemáticas de factores pronóstico para infección protésica.

BACKGROUND: Prosthetic joint infection is one of the most serious complications in orthopedics. Prognostic systematic reviews (SR) detecting and assessing factors related to prosthetic joint infection, allow better prediction of risk and implementation of preventive measures. Although prognostic SR are increasingly frequent, their methodological field presents some knowledge gaps. PURPOSE: To carry out an overview of SR assessing risk factors for prosthetic joint infection, describing and synthesizing their evidence. Secondarily, to assess the risk of bias and methodological quality. MATERIAL AND METHODS: We conducted a bibliographic search in 4databases (May 2021) to identify prognostic SR evaluating any risk factor for prosthetic joint infection. We evaluated risk of bias with the ROBIS tool, and methodological quality with a modified AMSTAR-2 tool. We computed the overlap degree study between included SR. RESULTS: Twenty-three SR were included, studying 15 factors for prosthetic joint infection, of which, 13 had significant association. The most frequently studied risk factors were obesity, intra-articular corticosteroids, smoking and uncontrolled diabetes. Overlapping between SR was high for obesity and very high for intra-articular corticoid injection, smoking and uncontrolled diabetes. Risk of bias was considered low in 8SRs (34.7%). The modified AMSTAR-2 tool showed important methodological gaps. CONCLUSIONS: Identification of procedural-modifiable factors, such as intra-articular corticoids use, can give patients better results. Overlapping between SR was very high, meaning that some SR are redundant. The evidence on risk factors for prosthetic joint infection is weak due to high risk of bias and limited methodological quality.

INTRAOCULAR EMULSION OF SILICONE OIL (ITEMS) GRADING SYSTEM: An Evidence-Based Expert-Led Consensus.

PURPOSE: To propose the InTraocular EMulsion of Silicone oil (ITEMS) grading system for the assessment of silicone oil (SiO) emulsion, applicable in a routine clinical setting and validated through an expert-led consensus procedure. METHODS: Seven experts on intraocular liquid tamponades, led by a facilitator, performed a literature review on the detection of SiO emulsion. Based on the proposed ideas, a questionnaire was developed and submitted to the experts on the methods to detect SiO emulsion and the items to grade. After 2 rounds of individual ranking using a 9-point scale and related discussion, the final grading system was developed including items that reached consensus (score ≥7 from ≥75% of members). RESULTS: The agreed ITEMS grading system includes the identification of SiO microbubbles and large SiO bubbles through slit-lamp biomicroscopy, gonioscopy, fundus examination under mydriasis, or ultra-wide-field fundus photography. Moreover, macular and disk optical coherence tomography are used to detect SiO-associated hyperreflective dots. CONCLUSION: An evidence-based expert-led consensus was conducted to develop grading system of SiO emulsion, allowing, for the first time, homogenous collection of data on SiO emulsion. This has the potential to improve the understanding of the role and clinical relevance of SiO emulsion, allowing comparisons between different studies.

[Translated article] Current situation of robotics in knee prosthetic surgery: A technology that has come to stay?

Robotic surgery is a surgical technique that is on the rise. The goal of robotic-assisted total knee arthroplasty (RA-TKA) is to provide the surgeon with a tool to accurately execute bone cuts according to previous surgical planning to restore knee kinematics and balance of soft tissue, being able to precisely apply the type of alignment that we choose. In addition, RA-TKA is a very useful tool for training. Within the limitations, there is the learning curve, the need for specific equipment, the high cost of the devices, the increase in radiation in some systems and that each robot is linked to a specific type of implant. Current studies show, with RA-TKA, variations in the alignment of the mechanical axis are reduced, postoperative pain is improved and earlier discharge is facilitated. On the other hand, there are no differences in terms of range of motion, alignment, gap balance, complications, surgical time or functional results.

Oncological monitoring after transanal total mesorectal excision (TaTME) for rectal neoplasia.

BACKGROUND: The surgical treatment of choice for rectal neoplasia is total mesorectal excision (TME). The transanal approach enables a better approach in male and obese patients and/or those with a narrow pelvis and in patients with small tumors. Short-term results are comparable with those for laparoscopy or the open approach, but the medium- and long-term oncological data are sparse. The aim of the present study was to evaluate our early experience with transanal TME (TaTME). METHODS: This was a retrospective study conducted on patients who underwent TaTME at our center between August 2013 and April 2017 with a follow-up ≥ 3 years. Histopathology, complications, mortality, neoplastic recurrence and disease-free survival were analyzed. RESULTS: One hundred patients (68 men and 32 women,, median age 66.8 years [range 29.6-91.2 years]) were included. There were 67 T3 cases (67%) with 74 N0 cases (74%), the mesorectal quality was graded optimal for 87.6% and only 2 cases of radial margin involvement were detected (2%). The median follow-up period was 47.6 months (range 11.8-78.9 months). Eighteen cases of recurrence were diagnosed, of which 3 (3%) recurred locally with an average disease-free period of 43.1 months. Overall survival was 80% and mortality due to progression of disease was 13%. CONCLUSIONS: TaTME is a safe surgical procedure with surgical, anatomopathological and oncological results at 3 years (medium-term) comparable with those for the laparoscopic and open approaches. Better monitoring is required with studies of the long-term functional and quality of life outcomes, i.e., at 5 or 10 years.

Current situation of robotics in knee prosthetic surgery, a technology that has come to stay? / Situación actual de la robótica en cirugía protésica de rodilla, ¿una tecnología que ha venido para quedarse?

Robotic surgery is a surgical technique that is on the rise. The goal of robotic-assisted total knee arthroplasty (RA-TKA) is to provide the surgeon with a tool to accurately execute bone cuts according to previous surgical planning to restore knee kinematics and balance of soft tissue, being able to precisely apply the type of alignment that we choose. In addition, RA-TKA is a very useful tool for training. Within the limitations, there is the learning curve, the need for specific equipment, the high cost of the devices, the increase in radiation in some systems and that each robot is linked to a specific type of implant. Current studies show, with RA-TKA, variations in the alignment of the mechanical axis are reduced, postoperative pain is improved and earlier discharge is facilitated. On the other hand, there are no differences in terms of range of motion, alignment, gap balance, complications, surgical time or functional results.

Ala®sil chemical characterization and toxicity evaluation: an example of the need for the Medical Device Regulation 2017/745.

Introduction: Ala®sil infusion was on the market for clinical use under the Medical Devices Directive (MDD) 93/42/EEC as an irrigating solution based on polydimethylsiloxane (PDMS). The product was withdrawn in 2016, and to the best of our knowledge, it did not cause any health damage. Methods: A bibliographic review and experimental analysis were conducted to evaluate whether this CE-marked product could have been used in patients under the current Medical Device Regulation (MDR) 2017/745. Analytical results from gas chromatography-mass spectrometry (GC-MS) and matrixassisted laser desorption ionization (MALDI) were performed. Citotoxicity studies were also carried out. Results: Only one study related to Ala®sil clinical use was found, describing a pilot series of five patients. The authors rated the product as not helpful in three out of the five cases for internal searching of retinal breaks and in four out of the five cases for drainage of subretinal fluid. No other scientific papers or documentation was found regarding Ala®sil's safety. Nevertheless, the product was introduced in the market after achieving the CE marking. GC-MS and MALDI showed that the polymer has a low molecular weight of 1,000 g/mol. Several linear and cyclic low-molecular-weight components (LMWCs) were identified as impurities ranging from L3 to D8, with a molecular weight below 600 g/mol. The Ala®sil sample was found to be cytotoxic after 24 h of cell culture but non-cytotoxic after 72 h, probably due to the cellular regeneration capacity of an immortalized cell line. Tissular cytotoxicity revealed an increased apoptosis rate but without morphological modifications. Discussion: Although Ala®sil cannot be classified as cytotoxic, this substance appears to increase retinal cell death processes. This study supports the notion that the MDDwas not functioning adequately to ensure the safety of medical devices. However, the current MDR 2017/745 imposes stricter standards to prevent the commercialization of medical devices without high-quality preclinical and clinical information, as well as precise clinical verification for their use, information not available for Ala®sil infusion.

[Towards a positive physiological definition of the deep brain nuclei in humans]. / Hacia una definición fisiológica positiva de los núcleos cerebrales profundos en humanos.

INTRODUCTION: Using microelectrodes for recording purposes in deep brain stimulation (DBS) has proven to be very useful. Their efficiency can be improved by characterising the properties of extracellular action potentials (EAPs). PATIENTS AND METHODS: We analysed the records of nine patients who underwent surgery for epilepsy or aggressiveness under general anaesthesia. The properties of the EAPs of the centromedian, ventral intermediate, ventrocaudal and posteromedial hypothalamic nuclei of the thalamus have been determined. RESULTS: We have analysed 706 thalamic and 142 hypothalamic cells. The proportion of cell types was found to be specific to each cell nucleus. The most frequent cell type was P1P2N1 (59.5%), followed by N1P1N2 (23.1%). The first phase of the EAP is highly variable. The properties of the EAP phases of the same morphology differ greatly from one nucleus to another. CONCLUSIONS: We have shown that several deep brain nuclei have properties that are specific to the morphology of the EAPs. This will allow for improved localisation of these nuclei during DBS.

TITLE: Hacia una definición fisiológica positiva de los núcleos cerebrales profundos en humanos.Introducción. El registro con microelectrodos en la estimulación cerebral profunda (ECP) ha demostrado una gran utilidad. Es posible mejorar su eficiencia caracterizando las propiedades de los potenciales de acción extracelulares (PAE). Pacientes y métodos. Hemos analizado registros de nueve pacientes operados por epilepsia o agresividad bajo anestesia general. Se han determinado las propiedades de los PAE de los núcleos talámicos centromediano, ventral intermedio, ventrocaudal e hipotalámico posteromedial. Resultados. Hemos analizado 706 células talámicas y 142 hipotalámicas. La proporción de tipos celulares resultó específica de cada núcleo celular. El tipo celular más frecuente fue P1P2N1 (59,5%), seguido por N1P1N2 (23,1%). La primera fase del PAE es altamente variable. Las propiedades de las fases del PAE de la misma morfología difieren altamente entre núcleos. Conclusiones. Hemos demostrado que diversos núcleos cerebrales profundos tienen propiedades específicas de la morfología de los PAE. Esto permitirá una mejora en la localización de estos núcleos durante la ECP.

Spray-driven halide exchange in solid-state CsPbX3 nanocrystal films.

CsPbI3 perovskite nanocrystals (NCs) are promising building blocks for photovoltaics and optoelectronics. However, they exhibit an essential drawback in the form of phase stability: α-phase, with a ∼1.80 eV bandgap, can easily experience a phase transition to a non-radiative orthorhombic δ-phase in an ambient environment. This leads to the need to carry out the CsPbI3-based device fabrication in an inert atmosphere, which is technologically inconvenient and expensive. One of the most successful approaches proposed to overcome this problem is synthesizing mixed halide CsPbBr3-xIx NCs to improve the stability of the α-phase perovskite structure. However, the formation of high-quality thin films of CsPbBr3-xIx NCs with high PLQY is challenging owing to the degradation of their optical properties after deposition on a substrate. This work presents spray coating to carry out a solid-state anion exchange in CsPbBr3 NCs thin films at ambient conditions with low-demanding reaction conditions. This constitutes a novel open-air and annealing-free technology to manufacture CsPbBr3-xIx NC thin films with high optical quality and record high photoluminescence quantum yields (PLQY) based on spray-driven halide (Br- to I-) anion exchange in a solid-state phase. Besides, tunable emission wavelengths between 520 and 670 nm can be obtained from CsPbBr3-xIx NC films using accurate tuning volumes of HI solution sprayed over the initial surface of CsPbBr3 film to provide the halide exchange. The optical quality of the halide-exchanged PNCs films remains practically identical to that of initial Br-containing layers, with a remarkable PLQY enhancement after anion exchange, from ∼61% for CsPbBr3 thin films emitting at 520 nm to ∼84% for mixed halide CsPbBr3-xIx film emitting at 640 nm. The huge potential of the system is confirmed by demonstrating a low-threshold amplified spontaneous emission.

Perioperative medicine role in painful knee prosthesis prevention.

Total knee arthroplasty is one of the most frequently performed orthopaedic surgeries. However, up to 20% of patients develop persistent postoperative pain. Persistent postoperative pain may be an extension of acute postoperative pain, but can also occur after more than 3 months without symptoms. Risk factors associated with persistent postoperative pain after arthroplasty have now been characterised within the patient's perioperative context (preoperative, intraoperative and postoperative), and can be grouped under genetic, demographic, clinical, surgical, analgesic, inflammatory and psychological factors. Identification and prevention of persistent postoperative pain through a multimodal and biopsychosocial approach is essential in the context of perioperative medicine, and has been shown to prevent or ameliorate postoperative pain.

Nanoplastics are bioaccumulated in fish liver and muscle and cause DNA damage after a chronic exposure.

The extent of the widespread, planetary contamination by plastic waste is difficult to fully capture. Nanoplastics (NPs) are currently in the center of research concerning plastic litter, both for the analytical challenges they pose and for their potential to provoke hazardous effects in organisms. However, there are still many unanswered questions in this multidisciplinary field, with a crucial missing piece being the quantification of NPs in fish tissues after in vivo exposures. Another relevant question that is still greatly unexplored is how a chronic exposure to NPs will affect fish health. This study aims to provide answers to both of these relevant knowledge gaps. To this end, goldfish (Carassius auratus) were exposed to 44 nm polystyrene (PS)-NPs via water for 30 days. Following the exposure, gastrointestinal tract, liver and muscle were sampled for PS-NPs analysis by means of size exclusion chromatography coupled to high resolution mass spectrometry. PS-NPs were detected in all liver and muscle samples of exposed fish, with higher concentrations in liver than in muscle, whereas no PS-NPs were detected in the gastrointestinal tract. Nevertheless, exposure to PS-NPs did not induce changes in hematology parameters nor in cortisol and glucose levels in plasma. On the other hand, even a relatively low concentration of PS-NPs was able to cause DNA damage, measured by an increase in erythrocyte nuclear abnormalities, suggesting that PS-NPs can reach the cell nucleus and cause genotoxicity. These results show for the first time that PS-NPs find their way to fish muscle after chronic exposure, where they bioaccumulate, but do not alter fish survival nor hematological or physiological stress indicators. The accumulation of PS-NPs in fish muscle can represent a threat to human health as a possible route of exposure to small-sized plastics. The present results in a model fish species open windows for future studies in edible fish species.

PREVENCOT project: Do we follow international guidelines to prevent surgical site infection in orthopedic elective surgery? / Proyecto PREVENCOT: ¿Seguimos las recomendaciones internacionales para la prevención de la infección del sitio quirúrgico en cirugía ortopédica programada?

BACKGROUND AND OBJECTIVE: Prevention of postoperative surgical site infections is indeed feasible. The aim of this work is to analyses adherence to international guidelines for the prevention of infections in elective orthopedic prosthetic surgery by means of a survey of a representative sample of Spanish orthopedic surgeons, with the purpose to establish general recommendations. MATERIAL AND METHOD: A population survey was conducted in online format consisting of 78 questions to analyze the usual clinical practice of Spanish orthopedic surgeons in the face of periprosthetic infections of the hip and knee, and their adherence to international guidelines. RESULTS: The results of the survey (n=138) show in clinical practice of Spanish orthopedic surgeons a high adherence to most of the international recommendations. CONCLUSIONS: The integration high adherence of individual clinical practice with the best available scientific evidence based on the recommendations of international guidelines is the best way to adequately manage the prevention of periprosthetic infection in elective surgery.

Hair cortisol level as a molecular biomarker in retinitis pigmentosa patients.

PURPOSE: Retinitis pigmentosa (RP) patients commonly experience negative psychological states due to their progressive and unpredictable loss of vision and visual variations related to stress. The aim of this study was to examine hair cortisol concentrations (HCCs), which is usually associated with chronic stress, pretending to unveil possible associations between underlying psychological factors and disease severity in RP patients. METHODS: Seventy-eight RP patients and 148 healthy controls were included in this study. A complete ophthalmological exam was performed in all patients to grade into severity disease groups. Perceived stress and trait-anxiety were measured by the State-Trait Anxiety Inventory (STAI) questionnaire. RESULTS: Fifty-two (67%) patients had severe RP and 26 (33%) mild-moderate RP. Fifty-eight (58,9%) patients reported severely levels of stress and 18 (23.,1%) highly levels assessed by STAI questionnaire. RP patients exhibited higher HCCs (500.04 ± 120.99 pg/mg) than in controls (136.17 ± 60.51 pg/mg; p < 0.001). Severe RP patients had significant higher HCCs than mild-moderate patients differing in 274.27 pg/mg (p < 0.001). RP severity grade and perceived anxiety levels in the questionaries were not associated. Group differences were not affected by relevant covariates (age, grade of severity, stress status, and gender). CONCLUSIONS: HCC seems an effective biomarker associated with chronic stress in RP patients. This study shows that HCC in patients with RP are elevated compared to population-based controls, and association between HCC and RP severity was found. Future research is needed to characterize the effect of untreated negative psychological states on progression of the disease if any.