Early stage breast cancer follow-up in real-world clinical practice: the added value of cell free circulating tumor DNA.

PURPOSE: Physical examinations and annual mammography (minimal follow-up) are as effective as laboratory/imaging tests (intensive follow-up) in detecting breast cancer (BC) recurrence. This statement is now challenged by the availability of new diagnostic tools for asymptomatic cases. Herein, we analyzed current practices and circulating tumor DNA (ctDNA) in monitoring high-risk BC patients treated with curative intent in a comprehensive cancer center. PATIENTS AND METHODS: Forty-two consecutive triple negative BC patients undergoing neoadjuvant therapy and surgery were prospectively enrolled. Data from plasma samples and surveillance procedures were analyzed to report the diagnostic pattern of relapsed cases, i.e., by symptoms, follow-up procedures and ctDNA. RESULTS: Besides minimal follow-up, 97% and 79% of patients had at least 1 non-recommended imaging and laboratory tests for surveillance purposes. During a median follow-up of 5.1(IQR, 4.1-5.9) years, 13 events occurred (1 contralateral BC, 1 loco-regional recurrence, 10 metastases, and 1 death). Five recurrent cases were diagnosed by intensive follow-up, 5 by symptoms, and 2 incidentally. ctDNA antedated disseminated disease in all evaluable cases excepted two with bone-only and single liver metastases. The mean time from ctDNA detection to suspicious findings at follow-up imaging was 3.81(SD, 2.68), and to definitive recurrence diagnosis 8(SD, 2.98) months. ctDNA was undetectable in the absence of disease and in two suspected cases not subsequently confirmed. CONCLUSIONS: Some relapses are still symptomatic despite the extensive use of intensive follow-up. ctDNA is a specific test, sensitive enough to detect recurrence before other methods, suitable for clarifying equivocal imaging, and exploitable for salvage therapy in asymptomatic BC survivors.

Blood-based genomics of triple-negative breast cancer progression in patients treated with neoadjuvant chemotherapy.

BACKGROUND: As neoadjuvant chemotherapy (NAC) is increasingly used in triple-negative breast cancer (TNBC), we investigated the value of circulating tumor DNA (ctDNA) for patient monitoring prior, during, and after NAC, and circulating tumor cells (CTCs) for disease characterization at clinical progression. MATERIALS AND METHODS: Forty-two TNBC patients undergoing NAC were prospectively enrolled. Primary tumor mutations identified by targeted-gene sequencing were validated and tracked in 168 plasma samples longitudinally collected at multiple time-points by droplet digital polymerase chain reaction. At progression, plasma DNA underwent direct targeted-gene assay, and CTCs were collected and analyzed for copy number alterations (CNAs) by low-pass whole genome sequencing. RESULTS: ctDNA detection after NAC was associated with increased risk of relapse, with 2-year event-free survival estimates being 44.4% [95% confidence interval (CI) 21.4%-92.3%] versus 77.4% (95% CI 57.8%-100%). ctDNA prognostic value remained worthy even after adjusting for age, residual disease, systemic inflammatory indices, and Ki-67 [hazard ratio (HR) 1.91; 95% CI 0.51-7.08]. During follow-up, ctDNA was undetectable in non-recurrent cases with the unique exception of one showing a temporary peak over eight samples. Conversely, ctDNA was detected in 8/11 recurrent cases, and predated the clinical diagnosis up to 13 months. Notably, recurrent cases without ctDNA developed locoregional, contralateral, and bone-only disease. At clinical progression, CTCs presented chromosome 10 and 21q CNAs whose network analysis showed connected modules including HER/PI3K/Ras/JAK signaling and immune response. CONCLUSION: ctDNA is not only associated with but is also predictive of prognosis in TNBC patients receiving NAC, and represents an exploitable tool, either alone or with CTCs, for personalized TNBC management.

Temporal relationship between successful pancreas transplantation and control of ocular complications in alloxan-induced diabetic rats.

PURPOSE: The impact of pancreas transplantation (PT) on the progression of eye disease is still controversial. This study evaluated the course of retinopathy in transplanted rats in two different diabetic stages. METHODS: Sixty inbred male Lewis rats were assigned to four experimental groups: NC-15 nondiabetic control rats; DC-15 untreated diabetic control rats; PT1-15 diabetic rats that received syngeneic pancreas transplants 2 weeks after alloxan diabetes induction; PT2-15 diabetic rats that received pancreas transplants 12 weeks after diabetes onset. Clinical and laboratory parameters and lens opacity were examined in all rats prior to treatment and at 1-, 6-, and 12-months follow-up. Nucleated eyes from five rats in each group processed for ultrastructural study of the retinal at 6 and 12 months after PT or at follow-up. RESULTS: Cataracts were observed in 20%, 60%, and 100% of DC rats at 1-, 6-, and 12-months follow-up, respectively. Early PT (2 weeks) significantly reduced the prevalence of this complication but not late (12 weeks) PT. PT1 rats also showed improved ultrastructure of the superficial and deep capillary plexuses of the retina, and of Müller cells, compared with DC and PT2. In the last group, retinopathy continued to evolve despite successful PT. CONCLUSION: Our results suggested that prevention of diabetic ocular lesions by PT was closely dependent on earlier performance of the procedure.

Pancreas transplantation prevents cellular oxidative stress in kidneys of alloxan-induced diabetic rats.

PURPOSE: Oxidative stress is one of the most important mechanisms to explain genesis of the complications in the chronic progression of diabetes. In this investigation we studied the effects of pancreas transplantation (PT) on the imbalance caused by excessive production of free oxygen radicals by antioxidant defenses of rats with serious chronic hyperglycemia induced by alloxan. METHODS: Ninety inbred male Lewis rats were randomly distributed into three groups: NC-30 nondiabetic controls; DC-30 diabetic controls without any treatment; PT-30 diabetic rats undergoing syngeneic PT from normal donor Lewis rats. Each experimental group was then split into three subgroups of 10 animals for sacrifice after 1, 3, or 6 months. Clinical and laboratory parameters from all rats as well as lipid hydroperoxide (LPO) concentrations and renal tissue enzyme activities of superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GSH-Px) were recorded for all rats. RESULTS: Successful PT corrected clinical and laboratory alterations in diabetic rats with sustained normoglycemia throughout the study. A significant increase in LPO concentration and a marked reduction in SOD and CAT enzyme activity were observed in DC rats; there was no significant variation in renal tissue GSH-Px in this group. However, alterations in DC rats were completely restored from 1st month after PT; all evaluated enzyme levels did not significantly differ (P < .01) from those in NC controls. CONCLUSION: Successful PT controlled cellular oxidative stress in diabetic kidneys, which may prevent chronic lesions.

Microscopic, morphometric and ultrastructural analysis of anastomotic healing in the intestine of normal and diabetic rats.

The purpose of this study was to investigate if experimental alloxanic diabetes could cause qualitative changes in intestinal anastomoses of the terminal ileum and distal colon in rats, as compared to controls. 192 male Wistar rats, weighing +/-300 g were split into four experimental groups of 48 animals each, after 3 months of follow-up: a control group with ileum anastomoses (G1), a control group with colon anastomoses (G2), a diabetic group with ileum anastomoses (G3) and a diabetic group with colon anastomoses (G4). Animals were evaluated and sacrificed on days 4, 14, 21 and 30 after surgery, and fragments of the small and large intestine where the anastomoses were performed were removed. Samples from 6 animals from each sacrifice moment were submitted to ultrastructural analysis of the collagen fibers using a scanning electron microscope and samples from another 6 animals were submitted to histopathology and optical microscopy studies using picrosirius red-staining. Histopathological analysis of picrosirius red-stained anastomosis slides using an optical microscope at 40x magnification showed that the distribution of collagen fibers was disarranged and also revealed a delay in scar tissue retraction. The morphometric study revealed differences in the collagen filled area for the ileum anastomoses 14 days post surgery whereas, in the case of colon anastomoses, differences were observed at days 4 and 30 post surgery, with higher values in the diabetic animals. Ultrastructure analysis of the ileum and colon anastomoses using a scanning electron microscope revealed fewer wide collagen fibers, the presence of narrower fibers and a disarranged distribution of the collagen fibers. We conclude that diabetes caused qualitative changes in scar tissue as well as in the structural arrangement of collagen fibers, what could explain the reduced wound strength in the anastomosis of diabetic animals.

CD157 is part of a supramolecular complex with CD11b/CD18 on the human neutrophil cell surface.

CD157 is a GPI-anchored cell surface glycoprotein expressed by human peripheral blood neutrophils. Cross-linking of CD157 induces intracellular Ca2+ mobilization and re-shaping in neutrophils, thus regulating their adhesive and migratory properties. Results obtained by immunolocalization and confocal microscopy indicate that CD157 lies in close proximity to the CD11b/CD18 complex which is strongly expressed on the activated neutrophil cell membrane where it plays a predominant role in adhesion. This study analyses the physical association between CD157 and CD18 in human neutrophils by co-immunoprecipitation experiments. The anti-CD157 monoclonal antibody RF3 co-precipitates CD18, and the anti-CD18 antibody TS1/18 co-precipitates CD157 from human neutrophil lysates. These results confirm that CD157 physically interacts with CD11b/CD18 complex in human neutrophils.

Demographic of short gut syndrome: increasing demand is not followed by referral of potential candidates for small bowel transplantation.

BACKGROUND: Despite improvements in small bowel transplantation (SBTx), early referral of patients with irreversible intestinal failure (IF) remains a major obstacle. In this study we evaluated the demand for SBTx among seven surgical pediatric centers located at least 200 km from our center. METHODS: From 1997 to 2001, 640 patients have been treated for neonatal diseases, including 248 who underwent a minor or major intestinal resection. Twenty-four patients with major resections presented with short gut syndrome, requiring total parenteral nutrition (TPN). The greatest demand was in postsurgical neonates with necrotizing enterocolitis, gastroschiesis, onphalocoeles, or midgut volvulus, and in three adults with postradiotherapy arteritis (n = 2) and mesenteric vein thromboses (n = 1). The median length of residual bowel after resection was 20 to 30 cm, without an ileocecal valve. Four patients were referred for SBTx evaluation; three died while awaiting a donor; 20 were not referred, among whom 14 died of TPN complications. RESULTS: Approximately 62 children per year require nutritional support for IF, most of whom develop complications related to TPN. Because many patients who are TPN-dependent develop complications, we believe that early referral would reduce mortality. CONCLUSIONS: Greater medical awareness about the feasibility of SBTx procedures and earlier referral may improve results and quality of life after transplant.

Anti-CD38 autoantibodies: characterisation in new-onset type I diabetes and latent autoimmune diabetes of the adult (LADA) and comparison with other islet autoantibodies.

AIMS/HYPOTHESIS: Serum anti-CD38 autoantibodies (aAbs) have been reported in 17 to 19% of patients with long-standing Type I (insulin-dependent) diabetes mellitus and Type II (non-insulin-dependent) diabetes mellitus. Whether these aAbs are also found in new-onset Type I diabetes and in Latent Autoimmune Diabetes in Adults (LADA) is not known, as is their relationship with conventional islet aAbs. METHODS: These issues were addressed by studying new-onset Type I and LADA diabetic cohorts with a recently developed anti-CD38 enzymatic immuno-assay. RESULTS: Anti-CD38 aAb prevalence among new-onset Type I patients (3.8%) was lower than previously found in long-standing Type I diabetes (11.7%, as defined with the 97.5 percentile cutoff; p=0.01), suggesting a late appearance of these aAbs. Among LADA patients, 14.9% were anti-CD38(+). Anti-CD38 were only associated with anti-GAD aAbs in new-onset Type I diabetes. Although the CD38 target molecule was expressed in human pancreatic islets, anti-CD38 aAbs did not contribute to the islet cell antibody (ICA) immunofluorescence reactivity. All the positive sera analysed for Ca(2+) release were found to mobilise it. In agreement with these agonistic features, anti-CD38(+) new-onset Type I patients showed higher fasting C-peptide values as compared to negative counterparts; the association was stronger when the analysis was limited to the agonistic anti-CD38(+) sera. A similar trend was found among LADA patients. CONCLUSION/INTERPRETATION: Anti-CD38 aAbs are distinct markers of islet autoimmunity which are more prevalent in long-standing disease, as opposed to the other known islet aAbs. Their in vitro agonistic properties could be operating in vivo as well, as they identify sub-groups of patients with higher residual beta-cell function.

Autoantibody response to CD38 in Caucasian patients with type 1 and type 2 diabetes: immunological and genetic characterization.

Insulin secretion is one of the functions mediated by CD38, a nonlineage pleiotropic cell surface receptor. The molecule is the target of an autoimmune response, because serum autoantibodies (aAbs) to CD38 have been detected in diabetic patients. In the healthy Caucasian population, the CD38 gene is bi-allelic (86% CD38*B and 14% CD38*A), whereas an Arg140Trp mutation has been identified in Japanese diabetic patients. We investigated the relationship between CD38 and diabetes in Caucasian patients by characterizing anti-CD38 aAbs in terms of prevalence and function (agonistic/nonagonistic activity) and by exploring the potential influence of the CD38 genetic background. A novel enzymatic immunoassay, using recombinant soluble CD38 as the target antigen, was developed for the analysis of anti-CD38 aAb titers. Sera from 19.15% of type 1 and 16.67% of type 2 diabetic patients were positive. The majority of anti-CD38 aAbs (57.14%) displayed agonistic properties, i.e., they demonstrated the capability to trigger Ca2+ release in lymphocytic cell lines. In agreement with these functional features, the presence of anti-CD38 aAbs in type 2 diabetic patients was associated with significantly higher levels of fasting plasma C-peptide and insulin, as compared with anti-CD38-counterparts. No diabetic subject carrying the Arg140Trp mutation and no preferential association between diabetes or aAb status and the CD38*A allele was found in the study population. These results show the significance of anti-CD38 aAbs as a new diagnostic marker of beta-cell autoimmunity in diabetes. Moreover, the prevalent agonistic activity of these aAbs suggests that they could mediate relevant effects on target cells by means of Ca2+ mobilization.

Identificación inmunohistoquímica de los fenómenos degenerativos en la hernia de disco lumbar: expresión de ácida hialurónico, metaloproteasas 1 y 3, factor de crecimiento fibroblástico básico y CD34 / Immunohistochemical detection of degenerative phenomea in lumbar disc herniations

Los objetivos de este trabajo son identificar los niveles de expresión y distribución de las metaloproteasas (MMP) 1 y 3, observar su efecto sobre el ácido hialurónico (AH), estudiar la respuesta inflamatoria, analizar los procesos de angiogénesis en el disco detectando la proteína CD34 y el factor de crecimiento fibroblástico básico (FCF-b) y relacionar los hallazgos con parámetros clínicos y epidemiológicos de los enfermos. Se analizaron discos intervertebrales obtenidos en 34 discectomías por hernia de disco y de 10 discos control con técnica histológica de rutina y técnicas inmunohistoquímicas contra MMP-1 y MMP-3, AH, CD34 y FCF-b. Se observó aumento significativo del porcentaje de degeneración, vascularización, detección de AH, MMP-1, MMP-3, CD34 y FCF-b en los discos extruidos y secuestros, lo que sugiere una relación entre las etapas más avanzadas de herniación y las más intensas de degeneración. Los enfermos con antecedentes de tabaquismo presentaron hernias con mayores niveles de expresión de FCF-b, o que sugiere una asociación entre el hábito de fumar y la intensidad de la respuesta tisular. Se discuten los hallazgos observados con una revisión de la literatura sobre el tema. La degeneración discal estaría producida por fenómenos biológicos que representan una falla en la capacidad de reparación del tejido conectivo especializado

Overdistraction of cervical spine injuries with the use of skull traction: a report of two cases.

Two cases in which cervical spine overdistraction occurred with the use of skull traction are described. A summary of the clinical presentations and definitive treatment together with some bibliographic references are discussed. Finally, suggestions regarding how to avoid overdistraction when using skull traction are given.

Giant sacral schwannoma. A case report.

STUDY DESIGN: Case report. OBJECTIVES: To present a rare case of a previously operated giant schwannoma located in the sacrum, and to make some considerations regarding diagnostic modalities and treatment options. SUMMARY OF BACKGROUND DATA: Large sacral schwannomas with anterior cortex erosion and associated intrapelvic extension are uncommon. Only a few case reports and small series have been published. There is no established consensus regarding diagnostic modalities, necessity for histologic diagnosis before surgery, or best surgical option. METHODS: The patient presented with a 2-month history of right sciatica and severe low back pain. After a histopathologic diagnosis and a complete set of image studies, the resection of the tumoral mass was planned posteriorly. RESULTS: Seventeen months after tumor resection, the patient has a good clinical outcome, and there are no radiologic signs of instability or recurrence. CONCLUSIONS: Considering the experience of the few cases reported in the world literature, the management of this tumor appears to grant favorable results, recurrence being the most frequent complication.

Canal estrecho lumbar / Lumbar spinal stenosis: diagnostic imaging

El proceso degenerativo del raquis lumbar es la causa principal de un trastorno clínico denominado Síndrome del canal estrecho lumbar. Varias hipótesis tratan de explicar este síndrome: la teoría postural y la teoría vascular-isquémica, aunque ambas pueden correlacionarse. Los síntomas y signos del canal estrecho son más frecuentes entre la sexta y octava década de vida, afectando a ambos sexos. El diagnóstico por imágenes juega un importante rol e incluye radiografías simples estáticas, TC, RM y la radiculografía dinámica. El canal estrecho puede ser central o de los recesos laterales y afecta con mayor frecuencia al cuarto espacio. Puede ser monofocal o polifocal. Su tratamiento es sintomático o quirúrgico, dependiendo de la gravedad de la estenosis y del cuadro neurológico. La casuística que presentamos corresponde a un período de 20 años con 150 pacientes intervenidos quirúrgicamente

La tomografía computada en los traumatismos vertebrales / The tomography computed in the vertebral injury

Durante mucho tiempo las radiografías simples y contrastadas y la tomografía lineal fueron los exámenes utilizados para el estudio de las fracturas vertebrales. En la actualidad la TC se ha constituido en un método esencial para juzgar el compromiso vertebral en los traumatismos que involucran el raquis, especialmente en aquellos casos que cursan un déficit neurológico. El uso de la TC requiere un conocimiento profundo por parte del radiólogo de la biomecánica de la columna, de los mecanismos y tipos de fracturas y de la técnica a aplicar. Presentamos nuestra experiencia sobre una población de 50 individuos (29 varones y 21 mujeres) con fracturas vertebrales. La edad promedio del conjunto fue 37,12 años con un rango etario comprendido entre 14 y 73 años. El grado de afectación neurológica medular se estableció según la grilla de Frankel. Entre las causas de los traumatismos tuvieron una alta incidencia los accidentes automovilísticos (37/50). Se identificaron 66 fracturas vertebrales (18 pacientes con 2 o mas fracturas), 14 luxaciones aisladas o combinadas con fracturas y 13 localizaciones de traumatismos extravertebrales. El mayor porcentaje de lesiones se produjo alrededor de la unión dorsolumbar y entre C4 y C7. La TC fue mas eficaz que las radiografías simples especialmente en las fracturas con trazo vertical y múltiples fragmentos y en el estudio del arco posterior incluyendo las facetas articulares y conducto raquídeo. Fueron de gran ayuda las reconstrucciones multiplanares