Intrapatient variation in PD-L1 expression and tumor mutational burden and the impact on outcomes to immune checkpoint inhibitor therapy in patients with non-small cell lung cancer.

BACKGROUND: PD-L1 tumor proportion score (TPS) and tumor mutational burden (TMB) are key predictive biomarkers for immune checkpoint inhibitors (ICI) efficacy in non-small cell lung cancer (NSCLC). Data on their variation across multiple samples are limited. METHODS: Patients with NSCLC and multiple PD-L1 TPS and/or TMB assessments were included. Clinicopathologic and genomic data were analyzed according to PD-L1 and TMB variation. RESULTS: In total, 402 PD-L1 sample pairs and 413 TMB sample pairs were included. Concordance between pairs was moderate for PD-L1 (ρ=0.53, P<0.0001) and high for TMB (ρ=0.80, P<0.0001). Shorter time between biopsies correlated with higher concordance in PD-L1, but not in TMB. Major increases (ΔTPS≥+50%) and decreases (ΔTPS≤-50%) in PD-L1 were observed in 9.7% and 8.0% of cases, respectively. PD-L1, but not TMB, decreased with intervening ICI (P=0.02). Acquired copy number loss of CD274, PDCD1LG2, and JAK2 were associated with major decrease in PD-L1 (q<0.05). Among patients with multiple PD-L1 assessments before ICI, cases where all samples had a PD-L1 ≥1%, compared to cases with at least one sample with PD-L1 <1% and another with PD-L1 ≥1%, achieved improved objective response rate and progression-free survival (PFS). Among patients with at least one PD-L1 <1% and one ≥1% before ICI, cases where the most proximal sample was PD-L1 ≥1% had longer median PFS compared to cases where the most proximal PD-L1 was <1%. Among patients with multiple TMB assessments before ICI, patients with a TMB ≥10 mut/Mb based on the most recent assessment, as compared to those with a TMB <10 mut/Mb, achieved improved PFS and OS to ICI; instead, no differences were observed when patients were categorized using the oldest TMB assessment. CONCLUSION: Despite intrapatient concordance in PD-L1 and TMB, variation in these biomarkers can influence ICI outcomes, warranting consideration for reassessment prior to ICI initiation when feasible.

Dissecting the clinicopathologic, genomic, and immunophenotypic correlates of KRASG12D-mutated non-small-cell lung cancer.

BACKGROUND: Allele-specific KRAS inhibitors are an emerging class of cancer therapies. KRAS-mutant (KRASMUT) non-small-cell lung cancers (NSCLCs) exhibit heterogeneous outcomes, driven by differences in underlying biology shaped by co-mutations. In contrast to KRASG12C NSCLC, KRASG12D NSCLC is associated with low/never-smoking status and is largely uncharacterized. PATIENTS AND METHODS: Clinicopathologic and genomic information were collected from patients with NSCLCs harboring a KRAS mutation at the Dana-Farber Cancer Institute (DFCI), Memorial Sloan Kettering Cancer Center, MD Anderson Cancer Center, and Imperial College of London. Multiplexed immunofluorescence for CK7, programmed cell death protein 1 (PD-1), programmed death-ligand 1 (PD-L1), Foxp3, and CD8 was carried out on a subset of samples with available tissue at the DFCI. Clinical outcomes to PD-(L)1 inhibition ± chemotherapy were analyzed according to KRAS mutation subtype. RESULTS: Of 2327 patients with KRAS-mutated (KRASMUT) NSCLC, 15% (n = 354) harbored KRASG12D. Compared to KRASnon-G12D NSCLC, KRASG12D NSCLC had a lower pack-year (py) smoking history (median 22.5 py versus 30.0 py, P < 0.0001) and was enriched in never smokers (22% versus 5%, P < 0.0001). KRASG12D had lower PD-L1 tumor proportion score (TPS) (median 1% versus 5%, P < 0.01) and lower tumor mutation burden (TMB) compared to KRASnon-G12D (median 8.4 versus 9.9 mt/Mb, P < 0.0001). Of the samples which underwent multiplexed immunofluorescence, KRASG12D had lower intratumoral and total CD8+PD1+ T cells (P < 0.05). Among 850 patients with advanced KRASMUT NSCLC who received PD-(L)1-based therapies, KRASG12D was associated with a worse objective response rate (ORR) (15.8% versus 28.4%, P = 0.03), progression-free survival (PFS) [hazard ratio (HR) 1.51, 95% confidence interval (CI) 1.45-2.00, P = 0.003], and overall survival (OS; HR 1.45, 1.05-1.99, P = 0.02) to PD-(L)1 inhibition alone but not to chemo-immunotherapy combinations [ORR 30.6% versus 35.7%, P = 0.51; PFS HR 1.28 (95%CI 0.92-1.77), P = 0.13; OS HR 1.36 (95%CI 0.95-1.96), P = 0.09] compared to KRASnon-G12D. CONCLUSIONS: KRASG12D lung cancers harbor distinct clinical, genomic, and immunologic features compared to other KRAS-mutated lung cancers and worse outcomes to PD-(L)1 blockade. Drug development for KRASG12D lung cancers will have to take these differences into account.

Antibiotic-exposed patients with non-small-cell lung cancer preserve efficacy outcomes following first-line chemo-immunotherapy.

BACKGROUND: Prior antibiotic therapy (pATB) is known to impair efficacy of single-agent immune checkpoint inhibitors (ICIs), potentially through the induction of gut dysbiosis. Whether ATB also affects outcomes to chemo-immunotherapy combinations is still unknown. PATIENTS AND METHODS: In this international multicentre study, we evaluated the association between pATB, concurrent ATB (cATB) and overall survival (OS), progression-free survival (PFS) and objective response rate (ORR) in patients with non-small-cell lung cancer (NSCLC) treated with first-line chemo-immunotherapy at eight referral institutions. RESULTS: Among 302 patients with stage IV NSCLC, 216 (71.5%) and 61 (20.2%) patients were former and current smokers, respectively. Programmed death-ligand 1 tumour expression in assessable patients (274, 90.7%) was ≥50% in 76 (25.2%), 1%-49% in 84 (27.9%) and <1% in 113 (37.5%). Multivariable analysis showed pATB-exposed patients to have similar OS {hazard ratio (HR) = 1.42 [95% confidence interval (CI): 0.91-2.22]; P = 0.1207} and PFS [HR = 1.12 (95% CI: 0.76-1.63); P = 0.5552], compared to unexposed patients, regardless of performance status. Similarly, no difference with respect to ORR was found across pATB exposure groups (42.6% versus 57.4%, P = 0.1794). No differential effect was found depending on pATB exposure duration (≥7 versus <7 days) and route of administration (intravenous versus oral). Similarly, cATB was not associated with OS [HR = 1.29 (95% CI: 0.91-1.84); P = 0.149] and PFS [HR = 1.20 (95% CI: 0.89-1.63); P = 0.222] when evaluated as time-varying covariate in multivariable analysis. CONCLUSIONS: In contrast to what has been reported in patients receiving single-agent ICIs, pATB does not impair clinical outcomes to first-line chemo-immunotherapy of patients with NSCLC. pATB status should integrate currently available clinico-pathologic factors for guiding first-line treatment decisions, whilst there should be no concern in offering cATB during chemo-immunotherapy when needed.

Reproducibility of peak power output during a 10-s cycling maximal effort using different sampling rates.

The study was aimed to investigate the reproducibility of performance parameters obtained from 10-s maximal cycling effort against different braking forces in young adult athletes. The sample (n = 48) included male athletes aged 18.9-29.9 years (175.5 ± 6.9 cm, 76.2 ± 10.1 kg). The exercise protocol was performed in a cycle-ergometer against a random braking force (4% to 11% of body mass). Intra-individual variation was examined from repeated tests within one week. Descriptive statistics were computed and differences between sessions were tested using paired t-test. The coefficient of correlation between repeated measures, technical error of measurement (TEM), coefficient of variation and ICC were calculated. Agreement between trials was examined using the Bland-Altman procedure. Mean values of peak power were relatively stable when obtained from sampling rates of 50 Hz and ranged between 1068 watt and 1082 watt (t(47) = 1.149, p = 0.256, ES-r = 0.165) or while corresponding to a sampling rate of 1 Hz (t(47) = 0.742, p = 0.462, ES-r = 0.107). Correlations between repeated measures were high (+0.907, 95% CI: +0.839 to +0.947) and TEM about 59.3 watt (%CV = 5.52%; ICC = 0.951, 95% CI: 0.912 to 0.972). The present study suggests that reproducibility of peak power in male adult athletes tended to be acceptable and within individual error appeared unrelated to braking force.

Maturity-associated variation in change of direction and dribbling speed in early pubertal years and 5-year developmental changes in young soccer players.

AIM: The purpose of the current study was to assess the developmental changes in change of direction and dribbling speed in youth soccer players taking into account skeletal age (SA), maturity status, body size, estimated fat mass, aerobic endurance, lower limb explosive strength and annual volume of training. METHODS: Eighty-three male soccer players aged 10-15 years (SA) at baseline were annually followed over 5 years, resulting in an average 4.4 observations per player. After testing for multicollinearity, multi-level regression modeling was used to examine the longitudinal developmental changes on change of direction and dribbling speed. RESULTS: Maturity-associated variability was significant in change of direction and also dribbling speed among young soccer players aged 12-14 years with better scores being performed by late maturers. Moreover, the predicted longitudinal scores for change of direction and dribbling speed improved with SA (P<0.01), SA2 (P<0.01) and skeletal maturity status entered as an additional developmental predictor (P<0.05). Estimated fat-free mass (P<0.01), aerobic endurance (P<0.01) and lower limb strength (P<0.01) were additional predictors in both models. The soccer-specific skill, dibbling speed, was also explained by annual volume of training (P<0.05). CONCLUSION: Skeletal maturity status explains inter-individual variability on maximal short-term run performances with and without the ball possession at early ages of participation in competitive soccer. The effects tend to persist across ages combined with longitudinal changes in body composition and functional fitness. In the particular case of the ball test, annual volume of training was also a longitudinal performance predictor.

Effects of habitat fragmentation on wild mammal infection by Trypanosoma cruzi.

Expansion of human activities frequently results in habitat fragmentation, a phenomenon that has been widely recognized in the last decades as one of the major threats to world's biodiversity. The transformation of a continuous forest into a fragmented area results in a hyper-dynamic landscape with unpredictable consequences to overall ecosystem health. The effect of the fragmentation process on Trypanosoma cruzi infection among small wild mammals was studied in an Atlantic Rain Forest landscape. Comparing continous forest to fragmented habitat, marsupials were less abundant than rodents in the continuous landscape. An overall decrease in small wild mammal richness was observed in the smaller fragments. An anti-T. cruzi seroprevalence of 18% (82/440) was deteced by immunofluorescence assay. Moreover, this seroprevalence was higher in the fragmented habitat than in the continuous forest. According to the collected data, 3 main factors seem to modulate infection by T. cruzi in small wild mammals: (i) habitat fragmentation; (ii) biodiversity loss; (iii) increase of marsupial abundance in mammal communities. Furthermore, an extremely mild controlled infection by T. cruzi was detected since no patent parasitaemia could be detected in fresh blood samples, and no parasites were isolated by haemoculture.

Mapping of the distribution of Trypanosoma cruzi infection among small wild mammals in a conservation unit and its surroundings (Northeast-Brazil).

Maps are a useful tool that permits correlation of landscapes with hotspots of parasite transmission. Here, they were used as a tool for geovisualization to evaluate variables involved in the transmission of Trypanosoma cruzi among small wild mammals in an area endemic for Chagas disease, the "Serra da Capivara" National Park (PARNA) and its surroundings in Piauí State, Northeast Brazil. The implementation of a Geographical Information System (GIS) allowed the observation that a previously noted aggregated distribution of Triatoma sordida and Triatoma brasiliensis, T. cruzi prevalence and infection pattern of small wild mammals was directly or indirectly influenced by the local relief and human action. Small mammalian species diversity was higher in mesic refugia inside the park and in its buffer zone and lower in the disturbed area by anthropic activities. Didelphis albiventris was more abundant in the areas affected by human action. Thrichomys laurentius demonstrated to be an eclectic species and a competent reservoir of T. cruzi, being infected in all study areas. Small wild mammals infected with the TCII genotype of T. cruzi were localized only in the buffer zone of PARNA while TCI infected specimens were found in both areas, inside the PARNA and its buffer zone. The impact of biodiversity loss on the transmission cycle of T. cruzi in the wild environment was discussed.

Clinical classification of tetanus patients.

The authors propose a clinical classification to monitor the evolution of tetanus patients, ranging from grade I to IV according to severity. It was applied on admission and repeated on alternate days up to the 10th day to patients aged > or = 12 years admitted to the State University Hospital, Recife, Brazil. Patients were also classified upon admission according to three prognostic indicators to determine if the proposed classification is in agreement with the traditionally used indicators. Upon admission, the distribution of the 64 patients among the different levels of the proposed classification was similar for the groups of better and worse prognosis according to the three indicators (P > 0.05), most of the patients belonging to grades I and II of the proposed classification. In the later reclassifications, severe forms of tetanus (grades III and IV) were more frequent in the categories of worse prognosis and these differences were statistically significant. There was a reduction in the proportion of mild forms (grades I and II) of tetanus with time for the categories of worse prognostic indicators (chi-square for trend: P = 0.00006, 0.03, and 0.00000) whereas no such trend was observed for the categories of better prognosis (grades I and II). This serially used classification reflected the prognosis of the traditional indicators and permitted the comparison of the dynamics of the disease in different groups. Thus, it becomes a useful tool for monitoring patients by determining clinical category changes with time, and for assessing responses to different therapeutic measures.

Clinical classification of tetanus patients

The authors propose a clinical classification to monitor the evolution of tetanus patients, ranging from grade I to IV according to severity. It was applied on admission and repeated on alternate days up to the 10th day to patients aged > or = 12 years admitted to the State University Hospital, Recife, Brazil. Patients were also classified upon admission according to three prognostic indicators to determine if the proposed classification is in agreement with the traditionally used indicators. Upon admission, the distribution of the 64 patients among the different levels of the proposed classification was similar for the groups of better and worse prognosis according to the three indicators (P > 0.05), most of the patients belonging to grades I and II of the proposed classification. In the later reclassifications, severe forms of tetanus (grades III and IV) were more frequent in the categories of worse prognosis and these differences were statistically significant. There was a reduction in the proportion of mild forms (grades I and II) of tetanus with time for the categories of worse prognostic indicators (chi-square for trend: P = 0.00006, 0.03, and 0.00000) whereas no such trend was observed for the categories of better prognosis (grades I and II). This serially used classification reflected the prognosis of the traditional indicators and permitted the comparison of the dynamics of the disease in different groups. Thus, it becomes a useful tool for monitoring patients by determining clinical category changes with time, and for assessing responses to different therapeutic measures.

Enzymatic determination of choline in milk using a FIA system with potentiometric detection.

The development of a FIA system for the determination of total choline content in several types of milk is described. The samples were submitted to hydrochloric acid digestion before injection into the system and passed through an enzymatic reactor containing choline oxidase immobilised on glass beads. This enzymatic reaction releases hydrogen peroxide which then reacts with a solution of iodide. The decrease in the concentration of iodide ion is quantified using an iodide ion selective tubular electrode based on a homogeneous crystalline membrane. Validation of the results obtained with this system was performed by comparison with results from a method described in the literature and applied to the determination of total choline in milks. The relative deviation was always < 5%. The repeatability of the method developed was assessed by calculation of the relative standard deviation (RSD) for 12 consecutive injections of one sample. The RSD obtained was < 0.6%.

Epidemiologia de las enfermedades del ganado bovino en Suriname / Epidemiology of cattle diseases in Suriname

The prevalence of certain cattle diseases in Suriname was studied over the period June-August 1985. The diseases were selected for their economic importance and association observed in the cattle with respiratory diseases, reproductive inefficiency, and neonatal mortality. Random samples were taken at two cattle-raising ranches, a dairy farm, and an abattoir in the District of Suriname. The study included a total of 478 animals, which were tested by indirect immunofluorescence for each of the diseases selected. Basically the serologic study revealed the presence of brucellosis, bovine viral diarrhea, parainfluenza 3, bovine infectious rhinotracheitis, and bovine coronavirus but not respiratory syncytial virus. The prevalence of parainfluenza 3 and bovine viral diarrhea was low compared with rates in other countries. It is recommended that a system, specially designed to meet the needs of this country, be established for the ongoing surveillance of data on animal health

Epidemiologic survey of bovine diseases in Suriname.

A seroepidemiologic survey of cattle diseases was undertaken in Suriname in 1985 to help assess the livestock disease situation in that country. The six diseases covered by the survey were bovine coronavirus infection, bovine rhinotracheitis, bovine virus diarrhea, brucellosis, parainfluenza-3 infection, and respiratory syncytial virus infection. The results indicated relatively low prevalences of these diseases compared to the prevalences found in most developed countries. The reasons for this are uncertain, but the finding suggests that the cattle population in Suriname could lack extensive exposure to these diseases and so could be highly susceptible to them. In addition, the evident need for more thoroughgoing survey data points up the need to establish a continuous animal data health monitoring system in Suriname--as well as in other developing countries where there is a need to objectively assess the livestock disease picture.

Epidemiologia de las enfermedades del ganado bovino en Suriname / Epidemiology of cattle diseases in Suriname

The prevalence of certain cattle diseases in Suriname was studied over the period June-August 1985. The diseases were selected for their economic importance and association observed in the cattle with respiratory diseases, reproductive inefficiency, and neonatal mortality. Random samples were taken at two cattle-raising ranches, a dairy farm, and an abattoir in the District of Suriname. The study included a total of 478 animals, which were tested by indirect immunofluorescence for each of the diseases selected. Basically the serologic study revealed the presence of brucellosis, bovine viral diarrhea, parainfluenza 3, bovine infectious rhinotracheitis, and bovine coronavirus but not respiratory syncytial virus. The prevalence of parainfluenza 3 and bovine viral diarrhea was low compared with rates in other countries. It is recommended that a system, specially designed to meet the needs of this country, be established for the ongoing surveillance of data on animal health