Evolving notions on immune response in colorectal cancer and their implications for biomarker development.

INTRODUCTION: Colorectal cancer (CRC) still represents the third most commonly diagnosed type of cancer in men and women worldwide. CRC is acknowledged as a heterogeneous disease that develops through a multi-step sequence of events driven by clonal selections; this observation is sustained by the fact that histologically similar tumors may have completely different outcomes, including a varied response to therapy. METHODS: In "early" and "intermediate" stage of CRC (stages II and III, respectively) there is a compelling need for new biomarkers fit to assess the metastatic potential of their disease, selecting patients with aggressive disease that might benefit from adjuvant and targeted therapies. Therefore, we review the actual notions on immune response in colorectal cancer and their implications for biomarker development. RESULTS: The recognition of the key role of immune cells in human cancer progression has recently drawn attention on the tumor immune microenvironment, as a source of new indicators of tumor outcome and response to therapy. Thus, beside consolidated histopathological biomarkers, immune endpoints are now emerging as potential biomarkers. CONCLUSIONS: The introduction of immune signatures and cellular and molecular components of the immune system as biomarkers is particularly important considering the increasing use of immune-based cancer therapies as therapeutic strategies for cancer patients.

Alagille syndrome case report: implications for forensic pathology and anthropology.

This case report offers a multidisciplinary interpretation of the violent death of a 4-year-old girl suffering from Alagille syndrome who died after a low-height fall that resulted in temporal bone fracture and a large epidural hematoma. The article evidences the macroscopical and microscopical characteristics of the syndrome, focusing especially on the skeletal findings that emerged during autopsy. In the case report, distinction is made between a possible accidental or non-accidental nature of the injuries and the characteristics of the injury have been interpreted in the light of the existing data on Alagille syndrome. In conclusion, the death was documented as accidental since abnormalities in the skeletal system evidenced during autopsy have predisposed the death of the child albeit through a very mild head trauma. The case report evidences the importance of studying features of skull macro- and microstructure in patients with Alagille syndrome, which have been, until now, underreported in literature and which might contribute to fracture vulnerability in these patients. Although rare, Alagille syndrome is a condition that should be known to forensic medicine practitioners and whose features and peculiarities must be taken into consideration in pediatric autopsy and suspected child abuse cases.

The carcinoid tumour in Meckel's diverticulum: how to treat young adults with appendicitis?--Case report.

This case reports the concomitant and unexpected finding of carcinoid tumour within a Meckel's diverticulum presenting as an acute abdomen due to gangrenous appendicitis in a young adult male. Both Meckel's diverticula and carcinoid tumour are rare clinical entities, and carcinoid tumours occurring within a Meckel's diverticulum are even more uncommon. The initial clinical presentation of carcinoid tumours occurring in a Meckel's diverticulum is usually similar to that of appendicle carcinoids.Carcinoid tumours are the most common primary tumour of the small bowel. They resemble appendicle carcinoids to the extent that they are usually small, single, and asymptomatic. Surgical treatment of Meckel's diverticula is recommended procedure. By presenting this case we wished to stress the value of systematic identification and removal of the diverticulum during laparotomy and to underline the importance of exploration the Meckel's diverticulum particularly in case of young adult subjects.

Neuroendocrine small cell carcinoma of the breast--a case report.

Neuroendocrine tumors are very rare tumors that occur most commonly in the gastrointestinal tract. The occurrence of neuroendocrine tumors outside gastrointestinal tract is very rare but not unknown. Thus, neuroendocrine tumors and their primary seat can be found in the bronchi and lungs, as well as in the testicles, ovaries, prostate, etc. The occurrence of neuroendocrine tumors as a primary seat in the breast is extremely rare phenomenon that is described in literature. We present the case of 55-year old female in where routine mammographic examination found suspicious lesions that we recommended for further processing. The patient made a breast ultrasound examination in which tumor formation was found in size 27 x 19 mm and cytological puncture found breast adenocarcinoma. Further pathohystologic and immunohistochemical analysis set the diagnosis of neuroendocrine carcinoma, small cell type, second grade. Tumor formation by ultrasound initially sized 27 x 19 mm and pathohistologic diagnosis showed tumor size 26 x 20 x 20 mm. The axillary lymph node biopsy did not found distant metastases in lymph nodes as well as gatherings in other organs. Neuroendocrine small cell carcinomas are exceedingly rare phenomena in the literature. By the year 2009 in the USA there were described only 50 cases of this extremely rare tumor of the breast.

Primary testicular necrotizing vasculitis clinically presented as neoplasm of the testicle: a case report.

We present a case of necrotizing vasculitis with the testicle as the isolated affected organ. A 25-year-old man, pretreated for epididymo-orchitis, presented with a presumed testicular neoplasm. Radical orchiectomy was performed and diagnosis of necrotizing vasculitis was established. In the absence of any other sign of systemic disease, the diagnosis of isolated necrotizing vasculitis of the testis was confirmed. Two years after the operation, the patient showed no symptoms of systemic disease.

The relationship between histologic chorioamnionitis and decidual macrophage polarization and their influence on outcomes of neonates born before the 32nd gestational week.

PURPOSE: To explore the relationship between histologic chorioamnionitis (HCA) and decidual macrophage (DM) polarization and their influence on outcomes of neonates born before the 32nd gestational week. MATERIALS AND METHODS: Eighty-four neonates and their placentas were included in this retrospective case-control study and divided into two groups: with and without HCA present (HCA + and HCA-). Neonatal, maternal, and placental risk factors were explored and their influence on neonatal outcomes was examined. We used CD68 and iNOS as markers for polarized DMs type 1 (M1) and CD163, CD206 and arginase (Arg-1) for polarized DMs type 2 (M2). RESULTS: HCA was present in 47 (56%) cases, and 37 (44%) cases were without the present HCA. There was no statistically significant difference in neonatal risk factors between the two groups (HCA + and HCA-). Higher rates of HCA (p = .042) were observed in mothers who received antepartum corticosteroid therapy. The frequency of vaginal deliveries in HCA + pregnancies was significantly higher than in HCA- pregnancies where deliveries by cesarean section were more frequently observed (p < .001). M2 DM were more abundant in the HCA + group (p = .035). Multiple regression model assessed the association between the presence of HCA, M1, and M2 DM with ROP stages. It has been observed that HCA is a risk factor for ROP stages (ß coefficient = 0.34, rpartial = 0.246, p = .024). With the logistic regression model, the association between the presence of HCA, M1, and M2 DM with neonatal nCPAP respiratory support and necrotizing enterocolitis (NEC) was assessed. The presence of M2 macrophages in decidua is an independent risk factor for neonatal nCPAP respiratory support (coefficient -0.07, OR = 0.928, 95% CI 0.87-0.99, p = .024) and the presence of M1 macrophages in decidua increases the risk for NEC (coefficient 0.010, OR = 1.0108, 95% CI 1.00-1.02, p = .032). CONCLUSIONS: The significantly more abundant presence of M2 DM was detected in HCA + placentas and their association with the increased risk for neonatal nCPAP respiratory support was observed. On the contrary, the presence of M1 DM increases the risk for NEC. The presence of HCA is a risk factor for ROP stages.

Dacryocystitis provoked by recurrence of extramedullary plasmacytoma of the orbit: a case report.

Extramedullary plasmacytoma (EMP) of the orbit is an extremely rare tumor, which frequently manifests with nonspecific symptoms. In the case reported, the symptoms of relapsing dacryocystitis appeared before the diagnosis of orbital EMP in a 60-year-old man. Moreover, EMP of the right submandibular gland that had been excised and treated by radiotherapy preceded the orbital lesion by seven years. The present report emphasizes the importance of an extensive medical workup to rule out multiple myeloma or other malignant lymphoproliferative diseases, because their treatment and prognosis are very different. Additionally, it is necessary to consider all tumors that can provoke relapsing dacryocystits.

Trauma related fat embolism syndrome in forensic practice.

The fat embolism syndrome (FES) in forensic practice is observed usually in cases of polytrauma related deaths. FES is rare, but serious complication after trauma. The most cases of post traumatic fat embolism are not fatal and it's very likely that many cases of mild fat embolism are overlooked. We describe a case of fat embolism syndrome in a young man after high energy injury. Upon concrete ceiling fell on his shoulders he had open fractures of both tibias and massive haematoma of the left hip joint. Transport immobilization of both legs was performed with admitting of analgesia during transportation to the hospital. Immediately after admission to hospital he underwent surgery. Several hours after osteosynthesis of both tibias, in ICU patient became hemodinamically and respiratory unstable. He developed global cyanosis and metabolic acidosis with significant decrease of oxygen pressure in the blood. Control chest radiograph showed typical "snow-storm" like pulmonary infiltrations. Deep bradicardy occurred followed by cardiopulmonary arrest. CPR was unsuccessful and the patient suddenly died within 12 hours from the incident. Histologic confirmation of fatty droplets the most commonly observed in the lung capillaries, brain capillaries or disseminated throughout the body remains diagnostic standard. We present one case of FES to emphasize the arising need of a quantitative analysis of both the size and localization of the fat emboli in order to grade the severity of FES and its relative contribution in pathophysiology of death. The postmortem diagnosis of fat embolism syndrome (FES), traditionally based on the histological demonstration of fat globules seems not to be enough, nowadays. A quantitative analysis of both the size and localization of the fat emboli has been discussed as reliable method of grading the pulmonary fat embolism in order to determine its relative functional contribution in death pathogenesis.

Atg5-Deficient Mice Infected with Francisella tularensis LVS Demonstrate Increased Survival and Less Severe Pathology in Internal Organs.

Francisella tularensis is a highly virulent intracellular pathogen that proliferates within various cell types and can infect a multitude of animal species. Francisella escapes the phagosome rapidly after infection and reaches the host cell cytosol where bacteria undergo extensive replication. Once cytosolic, Francisella becomes a target of an autophagy-mediated process. The mechanisms by which autophagy plays a role in replication of this cytosolic pathogen have not been fully elucidated. In vitro, F. tularensis avoids degradation via autophagy and the autophagy process provides nutrients that support its intracellular replication, but the role of autophagy in vivo is unknown. Here, we investigated the role of autophagy in the pathogenesis of tularemia by using transgenic mice deficient in Atg5 in the myeloid lineage. The infection of Atg5-deficient mice with Francisella tularensis subsp. holarctica live vaccine strain (LVS) resulted in increased survival, significantly reduced bacterial burden in the mouse organs, and less severe histopathological changes in the spleen, liver and lung tissues. The data highlight the contribution of Atg5 in the pathogenesis of tularemia in vivo.

Unveiling role of sphingosine-1-phosphate receptor 2 as a brake of epithelial stem cell proliferation and a tumor suppressor in colorectal cancer.

BACKGROUND: Sphingosine-1-phosphate receptor 2 (S1PR2) mediates pleiotropic functions encompassing cell proliferation, survival, and migration, which become collectively de-regulated in cancer. Information on whether S1PR2 participates in colorectal carcinogenesis/cancer is scanty, and we set out to fill the gap. METHODS: We screened expression changes of S1PR2 in human CRC and matched normal mucosa specimens [N = 76]. We compared CRC arising in inflammation-driven and genetically engineered models in wild-type (S1PR2+/+) and S1PR2 deficient (S1PR2-/-) mice. We reconstituted S1PR2 expression in RKO cells and assessed their growth in xenografts. Functionally, we mimicked the ablation of S1PR2 in normal mucosa by treating S1PR2+/+ organoids with JTE013 and characterized intestinal epithelial stem cells isolated from S1PR2-/-Lgr5-EGFP- mice. RESULTS: S1PR2 expression was lost in 33% of CRC; in 55%, it was significantly decreased, only 12% retaining expression comparable to normal mucosa. Both colitis-induced and genetic Apc+/min mouse models of CRC showed a higher incidence in size and number of carcinomas and/or high-grade adenomas, with increased cell proliferation in S1PR2-/- mice compared to S1PR2+/+ controls. Loss of S1PR2 impaired mucosal regeneration, ultimately promoting the expansion of intestinal stem cells. Whereas its overexpression attenuated cell cycle progression, it reduced the phosphorylation of AKT and augmented the levels of PTEN. CONCLUSIONS: In normal colonic crypts, S1PR2 gains expression along with intestinal epithelial cells differentiation, but not in intestinal stem cells, and contrasts intestinal tumorigenesis by promoting epithelial differentiation, preventing the expansion of stem cells and braking their malignant transformation. Targeting of S1PR2 may be of therapeutic benefit for CRC expressing high Lgr5.

Immunohistochemical surfactant protein-A expression: fatal drowning vs. postmortem immersion.

The postmortem diagnosis of drowning continues to be one of the most difficult in forensic pathology because of unspecific autopsy findings. It must be always remembered that disposal of a victim body in water is not unknown in homicide. The most important physiological consequence in fatal drowning is hypoxemia. The air-liquid interface of alveoli and distal airways of the mature lung are lined with a thin layer of lung surfactant, composed of phospholipids, proteins and neutral lipids. Surfactant components are synthesized and/or incorporated into lung surfactant in alveolar type II cells, and secreted to form an airspace lining film. The composition and function of lung surfactant is disturbed in cases od acute lung injury (ALI) including drowning. Surfactant protein-A (SP-A) is the most abundant surfactant protein. Surfactant protein-A (SP-A) is secreted by type II alveolar cells and cells. It's immunohistochemical distribution is observed in two different pathways: a linear membranous staining and a granular intra-alveolar staining. We hypothesize the significance of immunohistochemical detection of SP-A and its help in determination of the time of death, and possibly distinguishing of death by immersion vs. postmortem immersion using the morphological analysis applied on SP-A immunohistochemical stained lung tissue samples. We also argue in favor of routine use of SP-A staining in selected forensic cases where pathogenesis includes mechanical asphyxia and lung pathology. Although some studies reached conclusions to define the mechanisms involved in pathogenesis of mechanical asphyxia and aspiration necessity of additional studies arose. The mechanism of the production of massive aggregates remains to be determined. In the mean time the detection of SP-A (immunohistochemical) as well as biochemical is potentially useful tool in the forensic practice with possible application in daily practice.

Synergistic potential of Juniperus communis and Helichrysum italicum essential oils against nontuberculous mycobacteria.

OBJECTIVE: The present study evaluated the possible synergistic antimycobacterial interactions of Juniperus communis and Helichrysum italicum essential oils (EO). METHODS: Antimycobacterial potential was tested against Mycobacterium avium and Mycobacterium intracellulare using broth and water dilution method and checkerboard synergy method. Antiadhesion and antibiofilm effect of EOs was evaluated on biotic (HeLa cells) and abiotic surface (polystyrene). To evaluate the possible mechanisms of action, cellular leakage of proteins and DNA was tested and structural changes were visualized with a transmission electron microscope. RESULTS: MIC, minimum bactericidal concentration (MBC) and minimal effective concentration (MEC) were 1.6 mg ml-1 for J. communis EO and 3.2 mg ml-1 for H. italicum EO against both mycobacteria. All combinations of EOs in checkerboard synergy method produced fractional inhibitory concentration index values ranging from 0.501 to 1.5, corresponding to synergistic, additive or indifferent effects. Mycobacterium avium showed a greater tendency to create biofilm but these EOs at subinhibitory concentrations (sMIC) effectively blocked the adhesion and the establishment of biofilm. The exposure of both mycobacteria to MICs and sMICs lead to significant morphological changes: acquired a swollen form, ghost-like cell, disorganized cytoplasm detached from the cell wall. OD value of supernatant for both mycobacteria exposed to EOs have confirmed that there is a leakage of cellular material. CONCLUSION: The leakage of the cellular material is noticeably higher in sMIC, which is probably due to cell wall damage. sMIC of both EOs have an additive or synergistic effect, reducing MICs, limiting adhesion and preventing the formation of biofilms.

Severe Traumatic Brain Injury Induces Early Changes in the Physical Properties and Protein Composition of Intracranial Extracellular Vesicles.

Extracellular vesicles (EVs) are membranous nanostructures that can indicate undergoing processes in organs and thus help in diagnostics and prognostics. They are secreted by all cells, contained in body fluids, and able to transfer proteins, lipids and nucleic acids to distant cells. Intracranial EVs were shown to change their composition after severe traumatic brain injury (TBI) and therefore to have biomarker potential to evaluate brain events. Properties of intracranial EVs early after TBI, however, have not been characterized. Here, we assessed cerebrospinal fluid (CSF) up to seven days after isolated severe TBI for physical properties of EVs and their proteins associated with neuroregeneration. These findings were compared with healthy controls and correlated to patient outcome. The study included 17 patients with TBI and 18 healthy controls. EVs in TBI-CSF were visualized by electron microscopy and confirmed by immunoblotting for membrane associated Flotillin-1 and Flotillin-2. Using nanoparticle tracking analysis, we detected the highest range in EV concentration at day 1 after injury and significantly increased EV size at days 4-7. CSF concentrations of neuroregeneration associated proteins Flotillin-1, ADP-ribosylation Factor 6 (Arf6), and Ras-related protein Rab7a (Rab7a) were monitored by enzyme-linked immunosorbent assays. Flotillin-1 was detected solely in TBI-CSF in about one third of tested patients. Unfavorable outcomes included decreasing Arf6 concentrations and a delayed Rab7a concentration increase in CSF. CSF concentrations of Arf6 and Rab7a were negatively correlated. Our data suggest that the brain response within several days after severe TBI includes shedding of EVs associated with neuroplasticity. Extended studies with a larger number of participants and CSF collected at shorter intervals are necessary to further evaluate neuroregeneration biomarker potential of Rab7a, Arf6, and Flotillin-1.

Serum IL-6, IL-8, IL-10 and beta2-microglobulin in association with International Prognostic Index in diffuse large B cell lymphoma.

AIMS AND BACKGROUND: Diffuse large B-cell lymphoma displays striking heterogeneity at clinical, genetic and molecular levels. The International Prognostic Index is useful to predict the outcome of diffuse large B-cell lymphoma patients. However, patients with identical International Prognostic Index values in clinical practice exhibit marked variability in survival, suggesting the presence of significant residual heterogeneity within each category. Since cytokines such as interleukin-6, -8 and -10 play important roles in the pathogenesis of lymphomas, and plasma level of beta2-microglobulin is associated with the outcome of patients with diffuse large B-cell lymphoma, the aim of the present study was to determine whether these parameters combined with the International Prognostic Index would better stratify these patients to predict their prognosis. PATIENTS AND METHODS: The study included 46 untreated diffuse large B-cell lymphoma patients. RESULTS: All study parameters (International Prognostic Index, Ann Arbor stage, extranodal involvement, performance status, lactate dehydrogenase, beta2-microglobulin, interleukin-6 and -10, and response to therapy) except for patient age and serum interleukin-8 level were associated with overall survival. In addition, the International Prognostic Index was strongly correlated with beta2-microglobulin, interleukin-6, -8 and -10, and when combined these parameters significantly better stratified patients according to survival. On multivariate analysis, therapeutic response to the primary treatment, elevated interleukin-6 and -10 levels, and the International Prognostic Index were significant predictors of overall survival. CONCLUSIONS: Our data imply that interleukins and beta2-microglobulin evaluation should be used in association with the International Prognostic Index to define prognostic subgroups in diffuse large B-cell lymphoma patients.

Antimycobacterial potential of the juniper berry essential oil in tap water.

Mycobacterium avium complex-related diseases are often associated with poorly maintained hot water systems. This calls for the development of new control strategies. The aim of this study was to investigate the activity of essential oils (EOs) from the Mediterranean plants, common juniper, immortelle, sage, lavandin, laurel, and white cedar against Mycobacterium avium ssp. avium, Mycobacterium intracellulare, and Mycobacterium gordonae in culturing broth and freshwater as their most common habitat. To do that, we developed a new method of water microdilution to determine their minimal effective concentrations (MEC). The most active EO was the one from the common juniper with the MEC of 1.6 mg mL-1. Gas chromatography / mass spectrometry the juniper EO identified monoterpenes (70.54 %) and sesquiterpenes (25.9 %) as dominant component groups. The main monoterpene hydrocarbons were α-pinene, sabinene, and ß-pinene. The juniper EO significantly reduced the cell viability of M. intracellulare and M. gordonae at MEC, and of M. avium at 2xMEC. Microscopic analysis confirmed its inhibitory effect by revealing significant morphological changes in the cell membrane and cytoplasm of all three bacteria. The mode of action of the juniper EO on the cell membrane was confirmed by a marked leakage of intracellular material. Juniper EO has a great practical potential as a complementary or alternative water disinfectant in hot water systems such as baths, swimming pools, spa pools, hot tubs, or even foot baths/whirlpools.

Celiac disease: From pathophysiology to treatment.

Celiac disease, also known as "celiac sprue", is a chronic inflammatory disorder of the small intestine, produced by the ingestion of dietary gluten products in susceptible people. It is a multifactorial disease, including genetic and environmental factors. Environmental trigger is represented by gluten while the genetic predisposition has been identified in the major histocompatibility complex region. Celiac disease is not a rare disorder like previously thought, with a global prevalence around 1%. The reason of its under-recognition is mainly referable to the fact that about half of affected people do not have the classic gastrointestinal symptoms, but they present nonspecific manifestations of nutritional deficiency or have no symptoms at all. Here we review the most recent data concerning epidemiology, pathogenesis, clinical presentation, available diagnostic tests and therapeutic management of celiac disease.

VEGF expression is associated with negative estrogen receptor status in patients with breast cancer.

The aim of this study was to analyze the association between vascular endothelial growth factor (VEGF) expression on tumor cells and other clinicopathologic parameters in breast cancer that could give additional information on its prognostic significance. Immunohistochemical analysis of expression of VEGF, estrogen (ER) and progesterone receptor (PR), HER-2/neu, and Ki67 was performed in 233 breast cancers. VEGF expression estimated semiquantitatively was correlated with all the above-mentioned parameters as well as with clinicopathologic characteristics of breast cancer such as menopausal status of patients, tumor size, histologic and nuclear grade, vascular invasion, and lymph node status. Most of the tumor cells and some stromal components expressed VEGF. A higher percentage of VEGF-positive tumor cells was present in premenopausal patients and in ER-negative tumors. In postmenopausal patients tumors with a higher expression of VEGF were associated not only with ER-negative but also with HER-2/neu-positive tumor cells. These ER-negative tumors were characterized by a higher proliferative activity. Angiogenic switch as well as proliferative activity of breast cancer cells probably are unfavorably dependent on estrogen activity. This negative correlation between VEGF expression and ER status may not only shed more light on tumor biology but may also have future therapeutic implications.

The role of nuclear factor kappaB on angiogenesis regulation through monocyte chemotactic protein-1 in myeloma.

Multiple myeloma is malignant proliferation of plasma cells and plasmacytoid cells. Vascular endothelial growth factor (VEGF) is known to be one of the most important if not the main regulator of physiologic and pathologic angiogenesis which triggers growth, survival and migration of myeloma cells. It has been shown that circulating mature or bone marrow driven endothelial precursor cells play an important role in neovascularisation. In accordance with these observations, current therapeutic approaches to myeloma include VEGF inhibitors. Since angiogenesis inhibitors are heterogeneous in origin and potency, and their growing list includes many products with a different function it would be of benefit to determine the key molecule produced by transformed plasma cells which stimulates bone marrow environment to produce their homing "milieu" secreting different cytokines such as VEGF, IL-6, and monocyte chemotactic protein-1 (MCP-1). This molecule could be nuclear factor kappa B (NF-kappaB). It has been confirmed that myeloma cells express and produce NF-kappaB. It has been established recently that by blocking NF-kappaB production MCP-1 secretion is reduced up to 60%. If so, this would also reduce production of IL-6 and VEGF, since MCP-1 upregulates VEGF and IL-6 production. This way one could make bone marrow bad environment for myeloma cells to settle, followed with no disease progression. Targeting to NF-kappaB intended to inhibits its activation with receptor antagonist would possibly significantly inhibit lipopolysaccharide-induced IL-6, MCP-1 and TNF-alpha. All of them being stimulators for VEGF secretion and indirectly activation of angiogenesis. To conclude, angiogenesis could be induced by myeloma cells themselves through NF-kappaB activation pathway and by inhibiting its activation we might prevent myeloma expansion in bone marrow and progression of the disease by decreased MCP-1 secretion.