Damiano Caputo

Size and charge of nanoparticles following incubation with human plasma of healthy and pancreatic cancer patients

When nanoparticles (NPs) enter a biological environment, proteins bind to their surface forming a protein coating, which alters NP features giving it a biological identity, which controls its physiological response. The NP biological identity (size, charge and aggregation state) does strictly correlate with its physicochemical properties and the nature of the biological environment. While the former relationship has been extensively investigated, whether and how alterations in the physiological environment affect the biological identity of the NPs remains unclear. In this work we enrolled healthy and histologically proven pancreatic cancer patients. A statistically significant reduction in the level of clinically relevant proteins in cancer patients occurred. Positively and negatively charged lipid nanoparticles with two different surface chemistries (plain and PEGylated) were incubated with human plasma from both groups and characterized thoroughly by dynamic light scattering and zeta potential measurements. Only when plain positively charged NPs were tested, significant difference in zeta-potential between healthy and pancreatic cancer groups was found. This result implies that pooling human plasma from healthy volunteers might lead to a bias and thus unpredictable consequences in regard to previously optimized targeting profile.

Antimicrobial Susceptibility, Biochemical and Genetic Profiles of Staphylococcus haemolyticus Strains Isolated from the Bloodstream of Patients Hospitalized in Critical Care Units

Abstract Staphylococcus haemolyticus strains (n=20), responsible of blood stream infections, were consecutively isolated from patients hospitalized in two different wards at high risk of infection. Strains displayed high rate of resistance to oxacillin (90%). All strains but two with decreased susceptibility (MIC = 4 μg/mL), were sensitive to vancomycin. Ten strains were resistant to teicoplanin. Among the strains susceptible to glycopeptides, three displayed heteroresistance to vancomycin and seven to teicoplanin, when tested by Etest technique with 2 x McFarland inoculum. Biochemical reactions allowed to assign strains to eight biotypes, with 11 strains clustering under two main biotype A and biotype B. Pulsed-field-gel-electrophoresis (PFGE) identified 11 different PFGE-types. Seven strains grouping under the major PFGE-type 1 and three strains clustering in PFGE-type 2, closely correlated to biotype A and biotype B respectively. Seven teicoplanin-resistant isolates clustered in the PFGE-type 1, two in the PFGE-type 2 and one in PFGE-type 5. Therefore, teicoplanin-resistant strains were biochemically and genetically related and clonally distributed, despite different clones of S. haemolyticus circulated in the units during the study period.

Aberrant promoter methylation of beta-1,4 galactosyltransferase 1 as potential cancer-specific biomarker of colorectal tumors

Epigenetic alterations, such as CpG islands methylation and histone modifications, are recognized key characteristics of cancer. Glycogenes are a group of genes which epigenetic status was found to be changed in several tumors. In this study, we determined promoter methylation status of the glycogene beta‐1,4‐galactosyltransferase 1 (B4GALT1) in colorectal cancer patients. Methylation status of B4GALT1 was assessed in 130 colorectal adenocarcinomas, 13 adenomas, and in paired normal tissue using quantitative methylation specific PCR (QMSP). B4GALT1 mRNA expression was evaluated in methylated/unmethylated tumor and normal specimens. We also investigated microsatellite stability and microsatellite instability status and KRAS/BRAF mutations. Discriminatory power of QMSP was assessed by receiving operating curve (ROC) analysis on a training set of 24 colorectal cancers and paired mucosa. The area under the ROC curve (AUC) was 0.737 (95% confidence interval [CI]:0.591–0.881, P = 0.005) with an optimal cutoff value of 2.07 yielding a 54% sensitivity (95% CI: 35.1%–72.1%) and a specificity of 91.7% (95% CI: 74.1%–97.7%). These results were confirmed in an independent validation set where B4GALT1 methylation was detected in 52/106 patients. An inverse correlation was observed between methylation and B4GALT1 mRNA expression levels (r = −0.482, P = 0.037). Significant differences in methylation levels and frequencies was demonstrated in invasive lesions as compared with normal mucosa (P = 0.0001) and in carcinoma samples as compared with adenoma (P = 0.009). B4GALT1 methylation is a frequent and specific event in colorectal cancer and correlates with downregulation of mRNA expression. These results suggest that the glycogene B4GALT1 represent a valuable candidate biomarker of invasive phenotype of colorectal cancer.

Experience with two cases of intestinal tuberculosis: utility of the QuantiFERON-TB Gold test for diagnosis.

BACKGROUND Intestinal tuberculosis is rare in Western countries, with incidence rates of 35.7 and 0.43 per 100,000 per year for the immigrant and native populations, respectively. Despite a clear increase in the frequency of extrapulmonary tuberculosis in immunosuppressed patients, the clinical features of intestinal tuberculosis are seen rarely. A typical clinical presentation includes abdominal pain, weight loss, fever, weakness, nausea, vomiting, obstruction, and bleeding. Intestinal tuberculosis often mimics inflammatory bowel disease or malignant neoplasia, and its preoperative diagnosis is a challenge. Microbiologic diagnosis often is difficult because the causative microorganism requires a long incubation period. METHODS Two case reports and review of the pertinent literature. RESULTS We report two cases of colonic tuberculosis mimicking cecal carcinoma in one patient and periappendiceal abscess in the other. A 75 year-old man underwent right hemicolectomy for a right colon mass. Preoperative laboratory, radiologic, and endoscopic evaluations were negative for tuberculosis and carcinoma. The QuantiFERON-TB Gold test was positive. Surgical specimen histologic review showed non-caseating granulomas and rare Ziehl-Neelsen-positive bacilli. A 35 year-old man, born in Sri Lanka but living in Italy for 10 years, came to our attention for a periappendiceal abscess. Multiple peritoneal micro-nodules were found at laparotomy. Their extemporaneous histologic examination showed granulomas and giant-cell inflammation. A right hemicolectomy was performed. The QuantiFERON-TB Gold test, performed on peritoneal fluid and blood, was positive in both. CONCLUSIONS The QuantiFERON-TB Gold test may hold promise for use in intestinal inflammatory diseases when tuberculosis is suspected but conventional workup is not diagnostic.

Conversion in mini-invasive colorectal surgery: The effect of timing on short term outcome

INTRODUCTION Different results have been reported about postoperative outcomes of conversion during laparoscopic colorectal surgery. We aimed to detect the effect of conversion on postoperative outcome and to identify features associated to better outcome after conversion. METHODS Two hundred-fourteen mini-invasive left colonic and rectal resections were retrospectively analysed. Two groups were identified: mini-invasive colorectal surgery (MI) that includes both laparoscopic and robotic resections, and conversion to open surgery. RESULTS Among 214 colorectal procedures, 189 were MI. Conversion rate was 11.7%. Operating time was shorter for MI at overall analysis (p 0.003) and sub-analysis of left colectomies (p 0.001). MI procedures had shorter hospital stay (p 0.000) both in left colectomy and rectal resection (p 0.008 and p 0.001 respectively). A shorter time to first flatus emission was detected in MI group in both overall analysis (p 0.003) and procedures sub-analysis (left colectomy p 0.032; anterior rectal resection p 0.040). Oral feeding was resumed earlier after mini-invasive rectal resections (p 0.014). Converted procedures required more blood transfusions (p 0.000) and grade II complication rate was lower after MI procedures (p 0.013). Conversion presented higher anastomotic leakage and reoperation rates (p 0.035 and p 0.006 respectively). Conversion before 105 min (early conversion) had a significant lower number of blood transfusions (p 0.047). CONCLUSIONS Conversion is associated to higher rate of blood transfusions, grade II complication and slower recovery. Earlier conversion has better outcomes. Colorectal surgeons should identify any critical aspects that could avoid late conversion allowing reducing negative effects of conversion.

Neutrophil to Lymphocyte Ratio (NLR) and Derived Neutrophil to Lymphocyte Ratio (d-NLR) Predict Non-Responders and Postoperative Complications in Patients Undergoing Radical Surgery After Neo-Adjuvant Radio-Chemotherapy for Rectal Adenocarcinoma

ABSTRACT In order to evaluate neutrophil-to-lymphocyte ratio (NLR) and derived neutrophil-to-lymphocyte ratio (d-NLR) in predicting response and complications in rectal cancer patients who underwent surgery after neo-adjuvant radio-chemotherapy, 87 patients were evaluated. Cutoffs before and after radio-chemotherapy were respectively 2.8 and 3.8 for NLR, and 1.4 and 2.3 for d-NLR. They were analyzed in relation to clinical and pathological outcomes. Patients with preoperative NLR and d-NLR higher than cutoffs had significantly higher rates of tumor regression grade response (TRG ≥ 4) and postoperative complications. Elevated NLR and d-NLR after radio-chemotherapy are associated with worse pathological and clinical outcome.

T1 colorectal cancer: Poor histological grading is predictive of lymph-node metastases

INTRODUCTION After complete local excision of pT1 colorectal cancers, prediction of the absence of lymph-node involvement represents an interesting perspective in order to avoid unnecessary additional radical surgery, reducing morbidity, mortality and costs of care. We aimed to identify independent risk factors predictive of nodal involvement in pT1 colorectal cancer patients. METHODS Data regarding depth of submucosal invasion, histological grading, tumour budding and lymphovascular invasion in a consecutive series of 48 pT1 surgically resected colorectal cancers have been retrospectively collected and related to the nodal status. RESULTS A 12.5% rate of nodal involvement has been found. The poor differentiation was found as the only independent predictor of nodal metastases in pT1 colorectal cancer (p = 0.01). CONCLUSIONS Poor differentiation was the only independent significant predictor of nodal involvement in pT1 colorectal tumours. Our and literatures data confirm that risk factors must be prospectively collected and reported; further genetic and epigenetic predictive factors have to be investigated in order to carefully evaluate the needing of major surgery for pT1 colorectal cancer.

Phase II study of induction chemotherapy followed by chemoradiotherapy in patients with borderline resectable and unresectable locally advanced pancreatic cancer

There is not a clear consensus regarding the optimal treatment of locally advanced pancreatic disease. There is a potential role for neoadjuvant therapy to treat micrometastatic disease with chemotherapy, as well as for the treatment of local disease with radiotherapy. We evaluated the safety and efficacy of induction chemotherapy with oxaliplatin and gemcitabine followed by a high weekly dose of gemcitabine concurrent to radiation therapy in patients with borderline resectable and unresectable locally advanced pancreatic cancer. In our study, 41 patients with pancreatic cancer were evaluated. In all cases an accurate pre-treatment staging was performed. Patients with evidence of metastatic disease were excluded, and thus a total of 34 patients were consequently enrolled. Of these, twenty-seven patients (80%) had locally advanced unresectable tumours, seven patients (20%) had borderline resectable disease. This protocol treatment represents a well-tolerated promising approach. Fifteen patients (55.5%) underwent surgical radical resection. With a median follow-up of 20 months, the median PFS and OS were 20 months and 19.2 months, respectively. The median OS for borderline resectable patients was 21.5 months compared with 14 months for unresectable patients (p = 0.3). Continued optimization in multimodality therapy and an accurate patient selection remain crucial points for the appropriate treatment of these patients.

All that glitters are not gold! Reactive lymphoid hyperplasia mimicking colorectal liver metastases: description of a case and literature review

A 54-year-old woman with history of chronic thyroiditis because of changing in bowel habits with diarrhea underwent colonoscopy and a 6 cm polypoid tumor in the left colon was detected; biopsies showed a villous adenoma with high-grade dysplasia. Preoperative CT scan, laboratory, and CEA and Ca 19.9 levels were normal. Laparoscopic left colectomy was performed and a pT1N0G2 adenocarcinoma was detected. Liver ultrasonography performed 12 months later showed a nodular hypoechoic lesion of 9 mm in diameter between segment 5 and 8, without intravascular pattern (Fig. 1a). Hepatic enzymes were normal. MRI confirmed in the delayed phase, a suspicious hypo-intensive lesion with circular peripheral enhancement (Fig. 1b). Percutaneous needle biopsy showed reactive lymphoid infiltrates without neoplastic elements. CD3 positivity confirms the polyclonal reactive nature (T cells), CD20 positivity the presence of lymphoid B cells, and bcl-2 negativity in the germinal center confirms the reactive nature (Fig. 1c, d). Final pathological diagnosis was RLH. Close radiological follow-up was planned; 2 years after the diagnosis, there is no evidence of evolution neither changing in the dimensional characteristics of the hepatic lesion. RLH is a rare benign lesion that simulates malignancies (hepatocellular carcinoma, cholangiocarcinoma, and metastases). RHL mostly affects middle age women with single or multiple localizations. Lesion size ranges from 4 to 55 mm. Pathogenesis is unknown; more than half of cases are reported in autoimmune condition or chronic inflammation (i.e., autoimmune thyroiditis, viral hepatitis, or primary biliary cirrhosis). Diagnosis is generally occasional, natural history undefined and progressions to lymphoma in lungs, stomach and skin has been reported. The only low grade lymphoma arising from hepatic RLH in a 55-year-old patient, affected by primary biliary cirrhosis and Sjogren’s disease, was described in 1999. Since RLH is a benign condition, preoperative diagnosis is helpful to avoid useless resections. However, because of its radiological behavior with features similar to malignant lesions, non-invasive differential diagnosis is often challenging. Biopsy with immune-histochemical analysis is valuable to assess diagnosis; however, some authors do not agree on the sufficiency of biopsy in detecting low-grade lymphoma and suggest liver resection. However, considering the rarity of degeneration into lymphoma and that it is an indolent disease, in situ hybridization and analyses of gene rearrangements performed on sample from biopsy together with close radiological follow-up can be sufficient [1]. Thus, local or distant recurrences have been reported neither in resected cases nor in non-operated patients. To the best of our knowledge, to date, less than 15 cases of RLH have been reported in patients with oncologic history. The present is the sixth case of liver RLH in a patient with colon cancer history, and the first ever in which the lesion was detected during the early follow-up. & Damiano Caputo d.caputo@unicampus.it

Effect of Glucose on Liposome-Plasma Protein Interactions: Relevance for the Physiological Response of Clinically Approved Liposomal Formulations

Recently, the concept is emerging that the reduced success of nanoparticles in clinical practice is due to the adsorption of the “biomolecular corona (BC),” which alters their biological identity. Apart from protein variations, alterations in the human metabolome may change the BC decoration, which has poorly been addressed so far. Here, glucose is used as a model metabolite and how the interactions between liposomes (as a model nanoparticle) and plasma proteins are influenced by normal and diabetic sugar blood levels is explored. As model liposomes, Doxoves and Onivyde are used that are used for the treatment of breast and metastatic pancreatic cancer, respectively. It is shown that glucose does affect the structure and composition of BC. The biological effects of liposome–BC complexes are investigated in MCF 7 and MDA‐MB‐231 breast cancer cells for Doxoves and in pancreatic adenocarcinoma (PANC‐1) and insulinoma (INS‐1) cells for Onivyde. In the presence of glucose, the cellular toxicity of liposome–protein complexes and uptake by human monocytic THP1 cell line increases. These results demonstrate that alterations in glucose concentration, and more generally changes in the human metabolome, may play a fundamental role in the biological identity of liposomes and, consequently, on their in vivo physiological readouts including therapeutic efficacy.