Clinical Evolution Of Disease Research Articles

32P Can circulating immune checkpoints and KIT exon 11 mutations be prognostic factors in metastatic gastrointestinal stromal tumors?

32P Can circulating immune checkpoints and KIT exon 11 mutations be prognostic factors in metastatic gastrointestinal stromal tumors?

Vascular injury in macroscopically normal skin of patients with severe COVID-19 infection: clinical-pathologic correlations

Abstract Objectives: Taking into account that the documentation of the histopathological features in severe disease caused by SARS-CoV-2 has been scarce due to the avoidance of performing autopsies, the aim of the study was to detect the microscopic changes associated with severe COVID-19 infection in normal-appearing skin, without prominent dermatologic signs of a generalized microvascular thrombotic disorder, in accordance with the clinical evolution of disease. Methods: In this morphological and immunohistochemical study we included cutaneous biopsy samples from 12 symptomatic patients with severe and critical type SARS-CoV-2 infection (with the admission date between February and June 2020), treated in the Intensive Therapy Unit Care of Emergency County Hospital Targu-Mures, Romania. Results: The average age of our patients was 65.18 ± 14.21 years (range 41 to 83), and 66.67% of the patients were male. The histological and immunohistochemical assessment of cutaneous biopsies: in 4 cases the histological examination revealed small fibrin thrombi in deep-seated venules and small veins of subcuticular adipose tissue, and also 4 cutaneous biopsies showed occlusive vascular thrombosis in association with massive perivascular inflammatory infiltrate destroying and compromising the integrity of the vessel wall. The immunohistochemical examination of the composition of perivascular inflammatory infiltrate showed a predominance of CD3 positive lymphocytes, admixed with CD68 positive Mo/ MF, some of them activated with FXIII expression. In the perivascular infiltrate, the presence of granulocytes and B lymphocytes was not characteristic. Conclusion: According to our observations, in severe COVID-19, the cutaneous tissue is involved even in the absence of clinically obvious changes. Due to the relatively easy accessibility of skin samples, these could be applied to determine the severity of the patient’s clinical status, and to predict the necessity for anti-complement or anticoagulant treatments in the early stages of a severe SARS-CoV-2 infection.

Randomized, multicentre phase II trial of the sequencing of radium-223 and docetaxel plus prednisone in symptomatic bone-only metastatic castration-resistant prostate cancer (mCRPC).

TPS396 Background: Currently, the challenge is to reach a consensus on the best way to sequence different therapies for mCRPC patients (pts) in terms of efficacy and tolerability. Radium223 is a novel survival-prolonging targeted-α-therapy, but timing of its use in the treatment sequence of mCRPC remains an unanswered question, given the wide availability of therapeutic options Methods: This is a prospective, multicenter, randomized phase II study in symptomatic bone-only mCRPC pts who progressed after any androgen deprivation therapy, abiraterone and/or enzalutamide. Pts will be randomized 1:1 to receive radium223 initially followed by docetaxel+prednisone at the time of progression, or docetaxel+prednisone initially followed by radium223 at progression. In both treatment arms, the second step will be optional according to clinical evolution of disease (clinical deterioration and/or development of visceral metastases); however, each patient that will not enter in the second step will be still evaluable for the objectives of the study. No stratification factor will be used for randomization. Primary endpoint is to determine the better tolerated sequencing between radium223 and docetaxel in terms of health-related quality of life after completing the sequence and separately after each treatment. Based on primary endpoint, considering a type I error 0.1, type II error 0.2, proportion of responder pts in the standard arm 0.1 and in the experimental arm of 0.4, and assuming 10% loss to follow-up, a total of 70pts (35 for each arm) will be enrolled in the study. The study duration will be 36months; 15months of accrual, and 1 year of follow-up on the last participant enrolled. Secondary endpoints are to compare overall/progression-free/total progression free survival in pts treated with sequential therapy between radium223 and docetaxel. Additional secondary objective is to identify predictive factors for radium223 therapy, including potential circulating biomarkers through the plasma collection from all enrolled pts at different timepoints and the early prognostic role of functional imaging, such as PET with choline and/or PSMA. Clinical trial information: NCT03230734.

Evaluation of liquid biopsies for molecular profiling in patients with advanced non-small cell lung cancer (NSCLC) in the relapse treatment setting.

11532 Background: Molecular profiling is limited by access to sufficient tumor tissue for comprehensive analysis and due to tumor heterogeneity, the complete range of tumor DNA abnormalities may not be represented nor accurately reflect the clinical evolution of disease. Circulating tumor DNA (ctDNA) can be used as a minimally-invasive liquid biopsy for the detection, quantification and monitoring of molecular abnormalities for personalized treatment strategies. Methods: In a prospectively designed program, to date, we have recruited 227 advanced NSCLC patients having received prior therapy, with unknown molecular profile at time of blood collection. Blood collections (10ml K2-EDTA) were performed to assess molecular profile prior to or at time of relapse. Repeat samples were performed on patients initiated on treatment and followed for up to 18months. Patient samples were analyzed with InVision (enhanced tagged-amplicon sequencing) using a 34 gene panel. Interim analysis performed with full descriptive summary statistical analyses to be presented at conference. Results: ctDNA profiling detected somatic mutations in 182pts (80.2%), predominantly located in TP53 (46%), EGFR (28%), KRAS (11%) and STK11 (7%, half of which had concurrent KRAS). Of note, clinically actionable mutations were detected: T790M (25pts, median 1.4% AF), ERBB2 (8 pts), MET (8pts) and BRAF (4pts) providing eligibility for new therapy options. 20pts including 12 EGFR/T790M+ve were evaluated for ctDNA monitoring up to 18 months (median 10m); correlation between dynamic change in mutation allele fraction and clinical response was observed, especially predictive of relapse to treatment. 10 patients demonstrated SD/PR response to osimertinib treatment with T790M detection at low allele fraction (7/10 < 1% AF with 1pt at 0.08% AF). Conclusions: ctDNA can be used as a non-invasive ‘liquid biopsy’ for molecular profiling of NSCLC patients to detect clinically relevant and actionable mutations when tissue biopsy is unavailable. Liquid biopsies can be repeated as needed where tissue is not feasible, providing real-time information to support personalised treatment.

Evaluation of stored liquid biopsies for molecular profiling in patients with non-small cell lung cancer (NSCLC).

9024 Background: Molecular profiling is often limited by access to sufficient tumour tissue for comprehensive analysis and due to tumour heterogeneity, the complete range of tumor DNA abnormalities may not be represented nor accurately reflect the clinical evolution of disease. Circulating tumour DNA (ctDNA) can be used as a liquid biopsy for molecular abnormalities detection, quantification and monitoring for personalised treatment strategies. Methods: Plasma was collected at baseline (BL) and during study therapy from advanced NSCLC patients (pts) enrolled in a placebo controlled phase III trial of a novel irrerversible EGFR inhibitor; all patients had received standard therapy with chemotherapy and gefitinib or erlotinib. Archival tissue was collected when available but biopsy was not required prior to enrolment. BL Plasma ( < 3ml), stored for ~8 years was used to extract DNA and analysed using InVision (enhanced tagged-amplicon sequencing). Results: BL plasma from 387 pts was tested; 289 pts had available tissue results (from archival tissue collected at diagnosis) for EGFR (174WT/115Mut) and 243 for KRAS (205WT/38Mut). Despite age of plasma samples, ctDNA analysis detected cancer mutations in 310 pts (82%): TP53 (45%), KRAS (15%), and EGFR (43%; 56% were EGFR del19 and 29% L858R); T790M was detected in 80 patients. EGFR mutations were identified in 29 patients and KRAS in 10 patients with unknown tissue status. Also of note, STK11 (32 pts, 12 with KRAS), BRAF (5pts, 3 with V600E), MET(7 pts, 4 with METamp), ERBB2 (16pts, 10 with ERBB2amp) were identified in ctDNA analysis. Median time and median number of lines of systemic therapy between tissue biopsy and blood was 714 days and 3 lines respectively. Further analyses of ctDNA analyses in context of patient and trial outcomes are in progress. Conclusions: Liquid biopsies provide opportunity to evaluate molecular mutation profile in NSCLC patients. We demonstrate a highly sensitive method for ctDNA analysis which is complementary to tissue molecular analysis.

Biomarkers for Hepatocellular Apoptosis in the Management of Liver Diseases

Apoptosis is a rare event in normal hepatocytes. However, multiple signals can trigger apoptosis in hepatocytes and it plays a role in the pathogenesis of many liver diseases. This review summarizes the mechanisms of hepatocellular apoptosis and the importance of apoptosis in the pathological processes of liver disease. The potential for non-invasive biomarkers of apoptosis to gauge the extent and follow the evolution of clinical disease is emphasized.

Intérêt de la tomoscintigraphie cardiaque à la 123I-mIBG couplée à la perfusion myocardique dans le diagnostic de l’atrophie multisystématisée

Objective The aim of this prospective study is to assess the pertinence of using 123I-mIBG myocardial tomoscintigraphy coupled with perfusion scintigraphy as a diagnostic tool, to discriminate between multiple system atrophy (MSA) and idiopathic Parkinson's disease (PD) at first guided by clinical data and L-DOPA tests. Material and methods Forty patients, aged from 43 to 78 years (median 62 years) with Parkinson's syndrome were studied. Nineteen had a diagnosis of PD (criteria of brain bank) and 21 AMS (Gibbs criteria). All were given test to acute L-DOPA. Chest-centered planar imaging (128 × 128 matrix, 5 minutes of duration) is performed at 1 hour and 4 hours after injection of 220 MBq of 123I-mIBG, in addition a non-synchronised tomoscintigraphy (64 × 64 matrix, 32 images of 50 seconds , zoom 1.45) was performed after the 4th hour and 15 minutes after injection of 200 to 400 MBq of 99mTc-tetrofosmin. Besides neurological data, the parameters retained for comparison purposes with 123I-mIBG cardiac tomoscintigraphy were patients’ age, duration of disease and L-DOPA test results. Two regions of interest (ROI) identical in size and in shape are used for 123I-mIBG uptake quantifications (H/M and washout [Wo]). The first one was placed in projection of mediastinum (M) and the other one in projection of heart (H). Results We found an overall decreased uptake of the myocardial 123I-mIBG without perfusion abnormality in 15 of 19 patients with PD and 11 among them were L-DOPA sensitive (L-DOPA test greater than 30%). Normal tracer uptake with 123I-mIBG associated with an almost quite normal perfusion was seen in 15 of 21 patients with MSA and they were little or not L-DOPA sensitive (L-DOPA test less than 30%). Therefore, 10 discordant cases (25%) between cardiac scintigraphy and clinical evolution of disease with also discordant L-DOPA tests were observed. In the PD group, quantification of data enhanced the diagnostic decision with low heart to mediastinum ratio (H/M) (1.32 ± 0.15 at the early stage and 1.25 ± 0.13 at the later stage). In the MSA group, the uptake of 123I-mIBG (1.66 ± 0.43 at the early stage and 1.72 ± 0.42 at the later stage) was comparable to literature data, however, with significant inter-individual variations. The association of data of scintigraphy with L-DOPA test allows to improve sensitivity in 84% and specificity in 90.5%. Conclusion Our prospective study of 40 cases shows the relevance of cardiac sympathetic postganglionic imaging with 123I-mIBG coupled with myocardial perfusion scintigraphy to discriminate between MSA and PD with a higher sensitivity (71.4%) compared to the test with L-DOPA but a lower specificity (78.9%) than the L-DOPA. The difficulty of diagnosis is firstly linked to damage occurring to both the pre- and postganglionic sympathetic systems in patients with MSA and secondly to the integrity of the sympathetic nerve endings in patients with PD. However, the association of data of scintigraphy with L-DOPA test show a significant improvement of sensibility (84%).

ANTIRETROVIRAL THERAPY, VIRAL LOAD, AND T LYMPHOCYTES RATE IN HIV-1 INFECTED CHILDREN AFTER GENOTYPING DRUG RESISTANCE ASSESSMENT

Studies related to Human Immunodeficiency Virus type 1-infected children are of a special meaning due to multiple covariates such as timing of transmission, viral phenotypes, immunological patterns, viral dynamics progression and clinical evolution of disease. With antiretroviral therapy becoming more widely available, HIV resistance identification and monitoring of disease remains of great importance in infected children. The major HIV-1 infection markers usually used for monitoring viral infection and disease course are CD4+ T cell counts or percentages and HIV viral load. Both of them are helpful indicating when to start therapy and evaluating its efficacy. Also, their association with genotyping tests identifying viral resistant mutations may help clinicians for the most adequate clinical conduct. In the present study, we assessed HIV-1 viral load and CD4+ and CD8+ T lymphocyte rates for the immunological status evaluation of 25 antiretroviral-treated children or at the beginning of therapy, managing therapeutic regimens according to genotyping results. The management of highly active antiretroviral therapy (HAART) according to viral resistance in our group of pediatric patients allowed an increase in CD4+ T cell counts and/or percentage in almost all children, showing an improvement in their HIV-associated immunodeficiency status. Important viral burden declines were observed in 24 children, most of them multi-drug resistant, with HIV RNA undetectable levels reached in 12 of them. In particular, HAART introduction allowed a more significant viral load reduction for those pediatric patients who were drug treatment-naive, initiating antiretroviral therapy as they were enrolled at this study. DOI: http://dx.doi.org/10.17525/vrr.v14i1.23

Left ventricular hypertrophy: beyond the image and defining the human cardiac phenotype in hypertension

IntroductionLeft ventricular hypertrophy (LVH) has proven to be a very useful risk marker in hypertension. It indicates a higher risk of cardiovascular events and particularly sudden death [1,2]. It defines the effects of hypertension on an important target organ, the heart, and is one step along th

Influence of heme oxygenase 1 modulation on the progression of murine collagen‐induced arthritis

Heme oxygenase 1 (HO-1) can be induced by inflammatory mediators as an adaptive response. The objective of the present study was to determine the consequences of HO-1 modulation in the murine collagen-induced arthritis (CIA) model. DBA/1J mice were treated with an inhibitor of HO-1, tin protoporphyrin IX (SnPP), or with an inducer of HO-1, cobalt protoporphyrin IX (CoPP), from day 22 to day 29 after CIA induction. The clinical evolution of disease was monitored visually. At the end of the experiment, joints were examined for histopathologic changes. Cytokine levels in paws were measured by enzyme-linked immunosorbent assay. Levels of HO-1, cyclooxygenase 2 (COX-2), and prostaglandin E2 (PGE2) were determined. Effects of treatments on the early phase of disease and after prophylactic administration were also assessed. CoPP strongly induced HO-1, resulting in the inhibition of cartilage erosion accompanied by extensive fibrosis in the joint. Levels of tumor necrosis factor alpha (TNFalpha), interleukin-2 (IL-2), and IL-10 were inhibited by CoPP, whereas levels of vascular endothelial growth factor were increased. Treatment with SnPP significantly reduced the severity of CIA, with inhibition of joint inflammation and cartilage destruction. The levels of PGE2, IL-1beta, and TNFalpha were also significantly reduced by SnPP treatment, which did not modify COX-2 protein expression. SnPP was more effective than CoPP in preventing the development of CIA (prophylactic administration). HO-1 is induced during CIA. Although overexpression of this protein causes some beneficial effects, strategies aimed at HO-1 overexpression cannot slow the progression of the chronic inflammatory disease, whereas treatment with SnPP, which inhibits HO-1, exerts prophylactic and therapeutic effects.

Chronic hypoxia: common traits between chronic obstructive pulmonary disease and altitude.

Loss of body mass and exercise intolerance are common findings in chronic obstructive pulmonary disease and are often difficult to reverse despite optimal nutritional intake. Similar findings have been reported in healthy individuals during high-altitude exposure. The role of hypoxia in modulating metabolism has been largely investigated in vitro and in animal studies. More fragmentary is the knowledge regarding hypoxia effects on in-vivo human metabolism. This paper reviews recent literature regarding the effects of chronic exposure to hypoxia on metabolism, particularly comparing chronic obstructive pulmonary disease patients with humans exposed to high altitude. Hypoxia has important metabolic effects. Many oxygen-sensitive regulatory mechanisms work through hypoxia inducible factor 1, and recent literature regarding the hypoxic stimulus and its pathological implications deals largely with hypoxia inducible factor 1-related findings. Hypoxia inducible factor 1 is pivotal in the adaptation to chronic hypoxia: it induces gene expression for fructose-2-6-biphosphatase, an enzyme switching glucose metabolism towards glycolysis, allowing energy production in anaerobic conditions. Hypoxia inducible factor 1 is also involved in the development of anorexia because it induces the promoter of the leptin gene. Particularly important for future therapeutic implications are findings related to hypoxia inducible factor 1 polymorphism and interaction with other molecules, especially estrogens, in the clinical evolution of disease. Malnutrition is a worsening factor in chronic obstructive pulmonary disease. Similarities between chronic obstructive pulmonary disease and altitude exposure point to the importance of hypoxia in this regard. A better understanding of the underlying mechanisms will help to find alternative therapeutic approaches.

Three novel mutations reflect the variety of defects causing phenotypically diverse X-linked hyper-IgM syndrome.

X-linked hyper-IgM syndrome (HIGM1) (MIM musical sharp 308230), is a severe primary immunodeficiency caused by mutations in the gene coding for CD40 ligand (CD40L or CD154), a member of the tumour necrosis factor (TNF) superfamily. The interaction of this protein with its ligand, CD40, mediates crucial processes in the immune response. The variety of defects that have been described in HIGM1 patients range from a complete lack of CD40L protein expression to missense mutations that interfere with its interaction with CD40L. In this study we describe three families - a total of seven HIGM1 patients and carriers, presenting a spectrum of severity in clinical evolution. In two of these families, patient DNA samples were available for genetic studies. In the third, carrier detection was performed on female family members. The results of immunological studies - the different patterns of CD40L expression and binding capacity as measured by flow cytometry - and molecular diagnosis are presented. Three novel mutations were identified: an intron mutation that partially interferes with the splicing process (intron 3, position + 5 G/T); a missense mutation (Ser222 Phe) located in the molecular region which interacts with the receptor and which abrogates binding capacity; and a 14 base pair deletion leading to a frameshift and a premature truncated mutation (del I 171 X 195). An attempt to correlate protein expression and function of the CD40L mutants with clinical disease evolution is described.

Six years of experience in laparoscopic surgery of esophageal achalasia.

We show the experience of 6 years in laparoscopic surgical therapy of esophageal achalasia, performing of the Heller-Dor or Heller-Toupet operation, with particular regard to the technical aspect. One hundred and ten laparoscopic interventions were done between November 1995 and December 2001. We studied operative time in hiatus approach, esophagocardiomyotomy, transoperative endoscopy, and anti-reflux procedure. We also analyzed the relation between complications and clinical evolution of disease with clinical stage. The mean surgical time of the intervention is 138 min, and it is significantly increased by transoperative endoscopic control; conversion to open surgery was not necessary. Clinical results are classified as excellent in 103 patients. Morbidity was 6%. This surgical procedure is a first line in the treatment of esophageal achalasia. It is necessary to have special care in early diagnostic cases to avoid electrosurgical injury.

Vaccine protection against functional CTL abnormalities in simian human immunodeficiency virus-infected rhesus monkeys.

Accumulating evidence suggests that HIV-specific CD8(+) CTL are dysfunctional in HIV-infected individuals with progressive clinical disease. In the present studies, cytokine production by virus-specific CTL was assessed in the rhesus monkey model for AIDS to determine its contribution to the functional impairment of CTL. CTL from monkeys infected with nonpathogenic isolates of simian and simian-human immunodeficiency virus expressed high levels of IFN-gamma, TNF-alpha, and IL-2 after in vitro exposure to a nonspecific mitogen or the optimal peptide representing a dominant virus-specific CTL epitope. However, similarly performed studies assessing these capabilities in CTL from monkeys infected with pathogenic immunodeficiency virus isolates demonstrated a significant dysfunction in the ability of the CTL to produce IL-2 and TNF-alpha. Importantly, CTL from vaccinated monkeys that effectively controlled the replication of a highly pathogenic simian-human immunodeficiency virus isolate following challenge demonstrated a preserved capacity to produce these cytokines. These experiments suggest that defects in cytokine production may contribute to CTL dysfunction in chronic HIV or SIV infection. Moreover, an AIDS vaccine that confers protection against clinical disease evolution in this experimental model also preserves the functional capacity of these CTL to produce both IL-2 and TNF-alpha.

Evidence of increased class I MHC expression on human peripheral blood lymphocytes during acute ethanol intoxication.

Certain ethanol-related diseases in humans have been linked to disorders of immunity. Although humoral and cellular immunity have been studied, the precise mechanisms whereby ethanol use leads to tissue damage remain unknown. In order to explore the hypothesis that ethanol may lead to alteration in expression of tissue Class I major histocompatibility antigen causing an autoimmune phenomenon, a population of acutely ethanol-intoxicated patients was studied. Measurement of Class I major histocompatibility antigen on peripheral blood lymphocytes in this population showed a highly significant (p less than 0.01) increase over controls. The role that this increased antigenicity may play in the evolution of clinical disease is discussed.