Exploring kidney allograft rejection: A proof-of-concept study using spatial transcriptomics.

In this proof-of-concept study, spatial transcriptomics combined with public single-cell ribonucleic acid-sequencing data were used to explore the potential of this technology to study kidney allograft rejection. We aimed to map gene expression patterns within diverse pathologic states by examining biopsies classified across nonrejection, T cell-mediated acute rejection, interstitial fibrosis, and tubular atrophy. Our results revealed distinct immune cell signatures, including those of T and B lymphocytes, monocytes, mast cells, and plasma cells, and their spatial organization within the renal interstitium. We also mapped chemokine receptors and ligands to study immune cell migration and recruitment. Finally, our analysis demonstrated differential spatial enrichment of transcription signatures associated with kidney allograft rejection across various biopsy regions. Interstitium regions displayed higher enrichment scores for rejection-associated gene expression patterns than tubular areas, which had negative scores. This implies that these signatures are primarily driven by processes unfolding in the renal interstitium. Overall, this study highlights the value of spatial transcriptomics for revealing cellular heterogeneity and immune signatures in renal transplant biopsies and demonstrates its potential for studying the molecular and cellular mechanisms associated with rejection. However, certain limitations must be borne in mind regarding the development and future applications of this technology.

High-dimensional mass cytometry identified circulating natural killer T-cell subsets associated with protection from cytomegalovirus infection in kidney transplant recipients.

Cytomegalovirus (CMV) infection is associated with poor kidney transplant outcomes. While innate and adaptive immune cells have been implicated in its prevention, an in-depth characterization of the in vivo kinetics of multiple cell subsets and their role in protecting against CMV infection has not been achieved. Here, we performed high-dimensional immune phenotyping by mass cytometry, and functional assays, on 112 serially collected samples from CMV seropositive kidney transplant recipients. Advanced unsupervised deep learning analysis was used to assess immune cell populations that significantly correlated with prevention against CMV infection and anti-viral immune function. Prior to infection, kidney transplant recipients who developed CMV infection showed significantly lower CMV-specific cell-mediated immune (CMI) frequencies than those that did not. A broad diversity of circulating cell subsets within innate and adaptive immune compartments were associated with CMV infection or protective CMV-specific CMI. While percentages of CMV (tetramer-stained)-specific T cells associated with high CMI responses and clinical protection, circulating CD3+CD8midCD56+ NK-T cells overall strongly associated with low CMI and subsequent infection. However, three NK-T cell subsets sharing the CD11b surface marker associated with CMV protection and correlated with strong anti-viral CMI frequencies in vitro. These data were validated in two external independent cohorts of kidney transplant recipients. Thus, we newly describe the kinetics of a novel NK-T cell subset that may have a protective role in post-transplantation CMV infection. Our findings pave the way to more mechanistic studies aimed at understanding the function of these cells in protection against CMV infection.

Expression of Rejection-Associated Transcripts in Early Protocol Renal Transplant Biopsies Is Associated with Tacrolimus Exposure and Graft Outcome.

Subclinical inflammation in protocol biopsies relates to tacrolimus exposure and human leukocyte antigen (HLA) matching. We aimed to characterize transcripts associated with rejection and tacrolimus exposure and the latter's association with transplant outcomes. We tested whether gene expression is associated with rejection using strictly normal protocol biopsies (n = 17) and biopsies with T cell-mediated rejection (TCMR) or antibody-mediated rejection (ABMR) according to Banff criteria (n = 12). Subsequently, we analyzed these transcripts in a set of 4-month protocol biopsies (n = 137) to assess their association with donor and recipient characteristics, the intensity of immunosuppression, and the graft outcome. Differential expression (false discovery rate (FDR) < 0.01, fold (change (FC) > 3) between normal and rejection biopsies yielded a set of 111 genes. In the protocol biopsy cohort (n = 137), 19 out of these 111 genes correlated with tacrolimus trough levels at the time of biopsy (TAC-C0), and unsupervised analysis split this cohort into two clusters. The two clusters differed in donor age and tacrolimus trough levels. Subclinical rejection, including borderline lesions, tended to occur in the same cluster. Logistic regression analysis indicated that TAC-C0 at the time of biopsy (OR: 0.83, 95%CI:0.72-0.06, p = 0.0117) was associated with cluster 2. In a follow-up averaging 70 ± 30 months, this patient group displayed a significant decline in renal function (p = 0.0135). The expression of rejection-associated transcripts in early protocol biopsies is associated with tacrolimus exposure and a faster decline in renal function.

Tocilizumab en el tratamiento del rechazo humoral crónico activo resistente a terapia estándar / Tocilizumab in the treatment of active chronic humoral rejection resistant to standard therapy

Introducción: No existe consenso sobre el tratamiento más adecuado para el rechazo humoral crónico activo (RHCa). Estudios recientes sugieren que el tratamiento con tocilizumab (TCZ) puede estabilizar la función del injerto, disminuir la intensidad de los anticuerpos anti-HLA donante-específicos (ADEs) y reducir la inflamación de la microcirculación. Pacientes y métodos: Estudio observacional con pacientes trasplantados renales diagnosticados de RHCa (n = 5) que no habían presentado respuesta al tratamiento tradicional basado en la combinación de recambios plasmáticos, inmunoglobulinas y rituximab. A los pacientes se les indicó tratamiento con TCZ como uso compasivo en seis dosis mensuales (8 mg/kg/mes). Durante el seguimiento se monitorizó la función renal, proteinuria y la intensidad de los ADEs. Resultados: Cinco pacientes, de edad media 60 ± 13 años, tres de género masculino y dos retrasplantes (cPRA medio 55%) con ADEs preformados. El tratamiento con TCZ se inició a los 47 ± 52 días de la biopsia. En dos casos se suspendió el tratamiento tras la primera dosis, por bicitopenia severa con viremia por citomegalovirus y por fracaso del injerto, respectivamente. En los tres pacientes que completaron el tratamiento no se observó estabilidad de la función renal (creatinina sérica [Cr-s] de 1,73 ± 0,70 a 2,04 ± 0,52 mg/dL, filtrado glomerular estimado [FGRe] de 46 ± 15 a 36 ± 16 mL/min), presentaron aumento de la proteinuria (3,2 ± 4,0 a 6,9 ± 11,0 g/g) y la intensidad de los ADEs se mantuvo estable. No se observaron cambios en el grado de inflamación de la microcirculación (glomerulitis y capilaritis peritubular [g+cpt] 4,2 ± 0,8 vs. 4,3 ± 1,0), ni en el grado de glomerulopatía del trasplante (glomerulopatía crónica [cg] 1,2 ± 0,4 vs. 1,8 ± 1,0). (AU)

Introduction: There is no consensus on the most appropriate treatment for chronic active antibody-mediated rejection (cAMR). Recent studies suggest that treatment with tocilizumab (TCZ) may stabilize graft function, decrease the intensity of donor-specific HLA antibodies (DSAs) and reduce inflammation of microcirculation. Patients and methods: Observational study with renal allograft recipients diagnosed with cAMR (n = 5) who had not submitted a response to traditional treatment based on the combination of plasma replacements, immunoglobulins, and rituximab. Patients were told to be treated with TCZ as compassionate use in six doses per month (8 mg/kg/month). Renal function, proteinuria, and the intensity of DSAs were monitored during follow-up. Results: Five patients, average age 60 ± 13 years, three male and two retrasplants (cPRA average 55%) with preformed DSAs. Treatment with TCZ was initiated within 47 ± 52 days of biopsy. In two cases treatment was discontinued after the first dose, by severe bicitopenia with cytomegalovirus viremia and by graft failure, respectively. In the three patients who completed treatment, no stability of renal function (serum creatinine from 1.73 ± 0.70 to 2.04 ± 0.52 mg/dL, e-FGR 4 6 ± 15 to 36 ± 16 mL/min), showed increased proteinuria (3.2 ± 4.0 to 6.9 ± 11.0 g/g) and the intensity of DSAs maintain stable. No changes were observed in the degree of inflammation of microcirculation (g + pt 4.2 ± 0.8 vs. 4.3 ± 1.0) or in the degree of transplant glomerulopathy (cg 1.2 ± 0.4 vs. 1.8 ± 1.0). (AU)

Corrigendum: Enteric Budesonide in Transplant and Native IgA Nephropathy: Real-World Clinical Practice.

[This corrects the article DOI: 10.3389/ti.2022.10693.].

Tocilizumab in the treatment of active chronic humoral rejection resistant to standard therapy.

INTRODUCTION: There is no consensus on the most appropriate treatment for chronic active antibody-mediated rejection (cAMR). Recent studies suggest that treatment with tocilizumab (TCZ) may stabilize graft function, decrease the intensity of donor-specific HLA antibodies (DSAs) and reduce inflammation of microcirculation. PATIENTS AND METHODS: Observational study with renal allograft recipients diagnosed with cAMR (n = 5) who had not submitted a response to traditional treatment based on the combination of plasma replacements, immunoglobulins, and rituximab. Patients were told to be treated with TCZ as compassionate use in six doses per month (8 mg/kg/month). Renal function, proteinuria, and the intensity of DSAs were monitored during follow-up. RESULTS: Five patients, average age 60 ± 13 years, three male and two retrasplants (cPRA average 55%) with preformed DSAs. Treatment with TCZ was initiated within 47 ± 52 days of biopsy. In two cases treatment was discontinued after the first dose, by severe bicitopenia with cytomegalovirus viremia and by graft failure, respectively. In the three patients who completed treatment, no stability of renal function (serum creatinine from 1.73 ± 0.70 to 2.04 ± 0.52 mg/dL, e-FGR 4 6 ± 15 to 36 ± 16 mL/min), showed increased proteinuria (3.2 ± 4.0 to 6.9 ± 11.0 g/g) and the intensity of DSAs maintain stable. No changes were observed in the degree of inflammation of microcirculation (g+pt 4.2 ± 0.8 vs. 4.3 ± 1.0) or in the degree of transplant glomerulopathy (cg 1.2 ± 0.4 vs. 1.8 ± 1.0). CONCLUSIONS: TCZ therapy does not appear to be effective in modifying the natural history of chronic active antibody-mediated rejection, does not improve the degree of inflammation of microcirculation and does not reduces the intensity of DSAs.

Deciphering transplant outcomes of expanded kidney allografts donated after controlled circulatory death in the current transplant era. A call for caution.

Outcomes of kidney transplantation (KT) after controlled circulatory death (cDCD) with highly expanded criteria donors (ECD) and recipients have not been thoroughly evaluated. We analyzed in a multicenter cohort of 1161 consecutive KT, granular baseline donor and recipient factors predicting transplant outcomes, selected by bootstrapping and Cox proportional hazards, and were validated in a contemporaneous European KT cohort (n = 1585). 74.3% were DBD and 25.7% cDCD-KT. ECD-KT showed the poorest graft survival rates, irrespective of cDCD or DBD (log-rank < 0.001). Besides standard ECD classification, dialysis vintage, older age, and previous cardiovascular recipient events together with low class-II-HLA match, long cold ischemia time and combining a diabetic donor with a cDCD predicted graft loss (C-Index 0.715, 95% CI 0.675-0.755). External validation showed good prediction accuracy (C-Index 0.697, 95%CI 0.643-0.741). Recipient older age, male gender, dialysis vintage, previous cardiovascular events, and receiving a cDCD independently predicted patient death. Benefit/risk assessment of undergoing KT was compared with concurrent waitlisted candidates, and despite the fact that undergoing KT outperformed remaining waitlisted, remarkably high mortality rates were predicted if KT was undertaken under the worst risk-prediction model. Strategies to increase the donor pool, including cDCD transplants with highly expanded donor and recipient candidates, should be performed with caution.

Fraud Detection in Batches of Sweet Almonds by Portable Near-Infrared Spectral Devices.

One of the key challenges for the almond industry is how to detect the presence of bitter almonds in commercial batches of sweet almonds. The main aim of this research is to assess the potential of near-infrared spectroscopy (NIRS) by means of using portable instruments in the industry to detect batches of sweet almonds which have been adulterated with bitter almonds. To achieve this, sweet almonds and non-sweet almonds (bitter almonds and mixtures of sweet almonds with different percentages (from 5% to 20%) of bitter almonds) were analysed using a new generation of portable spectrophotometers. Three strategies (only bitter almonds, bitter almonds and mixtures, and only mixtures) were used to optimise the construction of the non-sweet almond training set. Models developed using partial least squares-discriminant analysis (PLS-DA) correctly classified 86-100% of samples, depending on the instrument used and the strategy followed for constructing the non-sweet almond training set. These results confirm that NIR spectroscopy provides a reliable, accurate method for detecting the presence of bitter almonds in batches of sweet almonds, with up to 5% adulteration levels (lower levels should be tested in future studies), and that this technology can be readily used at the main steps of the production chain.

COVID-19 in CKD Patients: Lessons from 553 CKD Patients with Biopsy-Proven Kidney Disease.

INTRODUCTION: Chronic kidney disease (CKD) patients infected with COVID-19 are at risk of serious complications such as hospitalization and death. The prognosis and lethality of COVID-19 infection in patients with established kidney disease has not been widely studied. METHODS: Data included patients who underwent kidney biopsy at the Vall d'Hebron Hospital between January 2013 and February 2020 with COVID-19 diagnosis during the period from March 1 to May 15, 2020. RESULTS: Thirty-nine (7%) patients were diagnosed with COVID-19 infection. Mean age was 63 ± 15 years and 48.7% were male. Hypertension was present in 79.5%, CKD without renal replacement therapy in 76.9%, and cardiovascular disease in 64.1%. Nasopharyngeal swab was performed in 26 patients; older (p = 0.01), hypertensive (p = 0.005), and immunosuppressed (p = 0.01) patients, those using RAS-blocking drugs (p = 0.04), and those with gastrointestinal symptoms (p = 0.02) were more likely to be tested for CO-VID-19. Twenty-two patients required hospitalization and 15.4% died. In bivariate analysis, mortality was associated with older age (p = 0.03), cardiovascular disease (p = 0.05), chronic obstructive pulmonary disease (p = 0.05), and low hemoglobin levels (p = 0.006). Adjusted Cox regression showed that low hemoglobin levels at admission had 1.81 greater risk of mortality. CONCLUSIONS: Patients with CO-VID-19 infection and kidney disease confirmed by kidney biopsy presented a mortality of 15.4%. Swab test for COVID-19 was more likely to be performed in older, hypertensive, and immunosuppressed patients, those using RAS-blocking drugs, and those with gastrointestinal symptoms. Low hemoglobin is a risk factor for mortality.

Kidney transplantation and COVID-19 renal and patient prognosis.

Coronavirus disease 2019 (COVD-19) emerged as a pandemic in December 2019. Infection has spread quickly and renal transplant recipients receiving chronic immunosuppression have been considered a population at high risk of infection, complications and infection-related death. During this year a large amount of information from nationwide registries, multicentre and single-centre studies have been reported. The number of renal transplant patients diagnosed with COVID-19 was higher than in the general population, but the lower threshold for testing may have contributed to its better identification. Major complications such as acute kidney injury and acute respiratory distress syndrome were very frequent in renal transplant patients, with a high comorbidity burden, but further studies are needed to support that organ transplant recipients receiving chronic immunosuppression are more prone to develop these complications than the general population. Kidney transplant recipients experience a high mortality rate compared with the general population, especially during the very early post-transplant period. Despite the fact that some studies report more favourable outcomes in patients with a kidney transplant than in patients on the kidney waiting list, the higher mortality described in the very early post-transplant period would advise against performing a kidney transplant in areas where the spread of infection is high, especially in recipients >60 years of age. Management of transplant recipients has been challenging for clinicians and strategies such as less use of lymphocyte-depleting agents for new transplants or anti-metabolite withdrawal and calcineurin inhibitor reduction for transplant patients with COVID-19 are not based on high-quality evidence.

An innovative non-targeted control system based on NIR spectral information for detecting non-compliant batches of sweet almonds.

Nowadays, there is growing awareness about the need to develop new methodologies to fight against deliberate fraud. This study explored the use of near infrared spectroscopy (NIRS) as an instantaneous, non-targeted method for detecting non-compliant products; in this case, when used to detect sweet almond batches adulterated with bitter almonds. For this purpose, we simulated the adulteration of batches by preparing four different types of mixed samples which contained 5%, 10%, 15% and 20% of bitter almonds, respectively, using 90 samples of sweet almonds and 50 samples of bitter almonds. For each of the adulteration percentages, 21 samples were produced. The samples were analysed using the Aurora and the Matrix-F spectrophotometers. The procedure initially constructed the desired standard or target using only the spectral information provided by the sweet almond population (control population). To achieve this, after principal components analysis, the spectral warning and action limits were calculated using the n-dimensional statistic Mahalanobis global distance. Next, the spectral distances from the product standard defined for those samples not belonging to the control population, including the adulterated sweet almonds, were calculated and represented as Shewhart control charts. The implementation of NIRS technology throughout the almond supply chain enabled to identify 87% (73/84) of the adulterated sweet almond batches. These findings suggest that NIRS technology and the use of spectral distances could enable to establish an innovative, non-targeted control system based only on spectral information to assess almond batches. This system allows to carry out conformity tests both in situ and online of the batches of almonds received and processed in the industry, as well as establishing fast, cost-efficient anti-fraud alert systems, which would help to reduce the number of batches to be analysed by expensive and time-consuming confirmatory methods.

Progression of Interstitial Fibrosis and Tubular Atrophy in Low Immunological Risk Renal Transplants Monitored by Sequential Surveillance Biopsies: The Influence of TAC Exposure and Metabolism.

The combination of tacrolimus (TAC) and mycophenolate is the most widely employed maintenance immunosuppression in renal transplants. Different surrogates of tacrolimus exposure or metabolism such as tacrolimus trough levels (TAC-C0), coefficient of variation of tacrolimus (CV-TAC-C0), time in therapeutic range (TTR), and tacrolimus concentration dose ratio (C/D) have been associated with graft outcomes. We explore in a cohort of low immunological risk renal transplants (n = 85) treated with TAC, mycophenolate mofetil (MMF), and steroids and then monitored by paired surveillance biopsies the association between histological lesions and TAC-C0 at the time of biopsy as well as CV-TAC-C0, TTR, and C/D during follow up. Interstitial inflammation (i-Banff score ≥ 1) in the first surveillance biopsy was associated with TAC-C0 (odds ratio (OR): 0.69, 95% confidence interval (CI): 0.50-0.96; p = 0.027). In the second surveillance biopsy, inflammation was associated with time below the therapeutic range (OR: 1.05 and 95% CI: 1.01-1.10; p = 0.023). Interstitial inflammation in scarred areas (i-IFTA score ≥ 1) was not associated with surrogates of TAC exposure/metabolism. Progression of interstitial fibrosis/tubular atrophy (IF/TA) was observed in 35 cases (41.2%). Multivariate regression logistic analysis showed that mean C/D (OR: 0.48; 95% CI: 0.25-0.92; p = 0.026) and IF/TA in the first biopsy (OR: 0.43, 95% CI: 0.24-0.77, p = 0.005) were associated with IF/TA progression between biopsies. A low C/D ratio is associated with IF/TA progression, suggesting that TAC nephrotoxicity may contribute to fibrosis progression in well immunosuppressed patients. Our data support that TAC exposure is associated with inflammation in healthy kidney areas but not in scarred tissue.

Routine NIRS analysis methodology to predict quality and safety indexes in spinach plants during their growing season in the field.

Cultural practices and harvesting in spinach plants should be based not only on subjective indexes such as freshness and green colour, which are both related with the visual appearance of the plants, but also on objective indexes that can be quantified non-destructively. The aim of this research was to develop a methodology based on the use of near infrared spectroscopy to monitor routinely the growth process of the spinach plants in the field. Using the MicroNIR™ OnSite-W spectrophotometer, which is ideally suited for in situ analysis, 261 spinach plants were analysed. Initially, calibration models for dry matter, soluble solid and nitrate contents were developed using 1 spectrum per plant for dry matter content, and nine spectra per plant for the other two parameters. These models were then validated using the same number of spectra per plant as for calibration purposes. After that, to establish a procedure more suitable to routine analysis in the field, the models were validated with only one spectrum per plant and the suitability of the predictions was measured considering the global and neighbourhood Mahalanobis distances, whose control limit values were defined as inferior to 4.0 and 1.0, respectively. The results showed that once the calibration models were developed, only one spectrum per plant was enough to predict dry matter and nitrate contents successfully. Therefore, the methodology developed will allow us to monitor in real time the complete growth process and to take decisions about spinach cultivation based on internal quality and safety indexes.

A Rejection Gene Expression Score in Indication and Surveillance Biopsies Is Associated with Graft Outcome.

Rejection-associated gene expression has been characterized in renal allograft biopsies for cause. The aim is to evaluate rejection gene expression in subclinical rejection and in biopsies with borderline changes or interstitial fibrosis and tubular atrophy (IFTA). We included 96 biopsies. Most differentially expressed genes between normal surveillance biopsies (n = 17) and clinical rejection (n = 12) were obtained. A rejection-associated gene (RAG) score was defined as its geometric mean. The following groups were considered: (a) subclinical rejection (REJ-S, n = 6); (b) borderline changes in biopsies for cause (BL-C, n = 13); (c) borderline changes in surveillance biopsies (BL-S, n = 12); (d) IFTA in biopsies for cause (IFTA-C, n = 20); and (e) IFTA in surveillance biopsies (IFTA-S, n = 16). The outcome variable was death-censored graft loss or glomerular filtration rate decline ≥ 30 % at 2 years. A RAG score containing 109 genes derived from normal and clinical rejection (area under the curve, AUC = 1) was employed to classify the study groups. A positive RAG score was observed in 83% REJ-S, 38% BL-C, 17% BL-S, 25% IFTA-C, and 5% IFTA-S. A positive RAG score was an independent predictor of graft outcome from histological diagnosis (hazard ratio: 3.5 and 95% confidence interval: 1.1-10.9; p = 0.031). A positive RAG score predicts graft outcome in surveillance and for cause biopsies with a less severe phenotype than clinical rejection.

Clinical characteristics and risk factors for severe COVID-19 in hospitalized kidney transplant recipients: A multicentric cohort study.

Kidney transplant recipients might be at higher risk for severe coronavirus disease 2019 (COVID-19). However, risk factors for relevant outcomes remain uncertain in this population. This is a multicentric kidney transplant cohort including 104 hospitalized patients between March 4 and April 17, 2020. Risk factors for death and acute respiratory distress syndrome (ARDS) were investigated, and clinical and laboratory data were analyzed. The mean age was 60 years. Forty-seven patients (54.8%) developed ARDS. Obesity was associated to ARDS development (OR 2.63; P = .04). Significant age differences were not found among patients developing and not developing ARDS (61.3 vs 57.8 years, P = .16). Seventy-six (73%) patients were discharged, and 28 (27%) died. Death was more common among the elderly (55 and 70.8 years, P < .001) and those with preexisting pulmonary disease (OR 2.89, P = .009). At admission, higher baseline lactate dehydrogenase (257 vs 358 IU/mL, P = .001) or ARDS conferred higher risk of death (HR 2.09, P = .044). In our cohort, ARDS was equally present among young and old kidney recipients. However, the elderly might be at higher risk of death, along with those showing higher baseline LDH at admission.

In situ ripening stages monitoring of Lamuyo pepper using a new-generation near-infrared spectroscopy sensor.

BACKGROUND: Near-infrared spectroscopy (NIRS) was used as a nondestructive sensor to assess the quality of freshly harvested Lamuyo peppers. One hundred and forty-four Lamuyo peppers, which were in a range of colors (green, chocolate, orange, and red) when harvested, were analyzed. In this study, the evolution of the main quality parameters during the harvest period was analyzed. Additionally, NIRS predictive models using a portable manual spectrophotometer to evaluate quality parameters together with color index were developed. Moreover, two procedures for taking near-infrared spectra were tested: (1) static, where point spectral readings were taken of around the equator of the fruit; (2) dynamic, where spectra were taken by scanning the entire length of the pepper. RESULTS: Green peppers and those harvested at the beginning of the campaign presented significantly lower values (P < 0.05) of dry matter, soluble solid contents, and titratable acidity, whereas those with red coloration and those harvested at the end of the campaign showed significantly higher values of these three quality parameters (P < 0.05). The predictive capacity of the NIRS models showed that the static mode proved to be the most suitable for measuring the quality of Lamuyo peppers. CONCLUSIONS: The viability of NIRS for measuring dry matter content and soluble solid contents in situ, using a new-generation NIRS sensor, was demonstrated. However, the high water content, the irregular shape of the fruit, and the fact that it is hollow inside all point to the need for using larger samples sets so as to increase the robustness of the models obtained. © 2019 Society of Chemical Industry.

Safety and quality issues in summer squashes using handheld portable NIRS sensors for real-time decision making and for on-vine monitoring.

BACKGROUND: Portable handheld near infrared spectroscopy (NIRS) instruments currently present enormous advantages in terms of size, weight, and robustness. They also provide fast, precise information that can be obtained in situ, and they represent a viable option for controlling vegetable safety and quality during the growth period. The aim of this research was to evaluate three handheld portable NIRS instruments for in situ and real-time analysis of intact summer squashes. Traditional methods were used to analyze 221 summer squashes, and this work was used to develop calibration models for morphological, safety, and quality parameters. The longitudinal distribution of nitrate content in summer squashes weighing over 400 g was also studied, and the evolution of this parameter during the harvest period was tracked to determine which summer squashes and which zones of the vegetables (peduncle, equatorial, or stylar) could be earmarked for baby-food production. RESULTS: The robustness of the calibration models confirmed the expectations raised by NIRS technology for morphological, safety, and quality control of individual summer squashes, and the models developed with the MicroNIR-1700 instrument were those that provided more accuracy and precision, being the peduncle zone the part with higher nitrate content. CONCLUSIONS: It is in the peduncle zone, therefore, where measurements of this parameter must be carried out to decide on the destination of the harvested product. Summer squashes picked at the end of the harvest are those that must be used for baby-food production. © 2019 Society of Chemical Industry.

LOCAL regression applied to a citrus multispecies library to assess chemical quality parameters using near infrared spectroscopy.

The non-destructive on-tree measurement of the chemical quality attributes of fruits belonging to the Citrus genus using rapid spectral sensors is of vital interest to citrus growers, allowing them to carry out a selective harvest of any species of Citrus fruit. With this objective, the viability of using of a handheld portable near infrared spectroscopy (NIRS) instrument to predict soluble solid content (SSC), pH, titratable acidity (TA), maturity index and BrimA, in order to measure the optimum harvest time in a group made up of 608 samples belonging to the Citrus genus (378 oranges and 230 mandarins) was evaluated. For each of the parameters analysed, both non-linear regression (LOCAL algorithm) and linear regression (Modified Partial Least Squares, MPLS) strategies were designed and compared. The use of the LOCAL algorithm in the sample group of oranges and mandarins for all the parameters analysed allowed to obtain more robust models than those obtained with MPLS regression, and it could also be extended more easily when routinely applied. The results confirm that NIRS technology combined with non-linear regression strategies such as the LOCAL algorithm can indeed respond to the needs of the Citrus growers and help them to set the optimum harvest time, in this case of oranges and mandarins, by predicting the chemical quality parameters in situ.

Rapid, simultaneous, and in situ authentication and quality assessment of intact bell peppers using near-infrared spectroscopy technology.

BACKGROUND: The ability of near-infrared (NIR) spectroscopy to authenticate individual bell peppers as a function of the growing system (outdoor or greenhouse) was tested using partial least squares discriminant analysis. Bell peppers grown outdoors (130 samples) or in a greenhouse (264 samples) during the 2015 and 2016 seasons were selected for this purpose and analysed using a portable, handheld, microelectromechanical system (MEMS) instrument MicroPhazir (spectral range 1600-2400 nm), working in reflectance. Subsequently, the potential of NIR spectroscopy as a non-destructive sensor for in situ quality (dry matter and soluble solid content) measurements, was investigated. RESULTS: The models correctly classified 89.73% and 88.00% of the samples by growing system, when trained with unbalanced and balanced sets respectively, mainly due to the differences in physical-chemical attributes between bell peppers cultivated in the two growing systems. Separate classification models for bell peppers grouped by ripeness (judged by the colour), allowed the classification of 88.28-91.37% of the samples correctly. The standard error of cross-validation values for the quantitative models were 0.66% fresh weight and 0.75 °Brix for dry matter and soluble solid content, respectively. CONCLUSIONS: The results showed that NIR spectroscopy can be used successfully for predicting the growing systems used in bell pepper production, which is of particular value to guarantee the authentication of outdoor-grown peppers. Additionally, the results showed that NIR spectroscopy can be used simultaneously as a rapid preliminary screening technique to measure quality. © 2018 Society of Chemical Industry.