Rectal Tract Research Articles

Automated Prediction of the Response to Neoadjuvant Chemoradiotherapy in Patients Affected by Rectal Cancer.

Simple SummaryColorectal cancer is the second most malignant tumor per number of deaths after lung cancer and the third per number of new cases after breast and lung cancer. The correct and rapid identification (i.e., segmentation of the cancer regions) is a fundamental task for correct patient diagnosis. In this study, we propose a novel automated pipeline for the segmentation of MRI scans of patients with LARC in order to predict the response to nCRT using radiomic features. This study involved the retrospective analysis of T2-weighted MRI scans of 43 patients affected by LARC. The segmentation of tumor areas was on par or better than the state-of-the-art results, but required smaller sample sizes. The analysis of radiomic features allowed us to predict the TRG score, which agreed with the state-of-the-art results.Background: Rectal cancer is a malignant neoplasm of the large intestine resulting from the uncontrolled proliferation of the rectal tract. Predicting the pathologic response of neoadjuvant chemoradiotherapy at an MRI primary staging scan in patients affected by locally advanced rectal cancer (LARC) could lead to significant improvement in the survival and quality of life of the patients. In this study, the possibility of automatizing this estimation from a primary staging MRI scan, using a fully automated artificial intelligence-based model for the segmentation and consequent characterization of the tumor areas using radiomic features was evaluated. The TRG score was used to evaluate the clinical outcome. Methods: Forty-three patients under treatment in the IRCCS Sant’Orsola-Malpighi Polyclinic were retrospectively selected for the study; a U-Net model was trained for the automated segmentation of the tumor areas; the radiomic features were collected and used to predict the tumor regression grade (TRG) score. Results: The segmentation of tumor areas outperformed the state-of-the-art results in terms of the Dice score coefficient or was comparable to them but with the advantage of considering mucinous cases. Analysis of the radiomic features extracted from the lesion areas allowed us to predict the TRG score, with the results agreeing with the state-of-the-art results. Conclusions: The results obtained regarding TRG prediction using the proposed fully automated pipeline prove its possible usage as a viable decision support system for radiologists in clinical practice.

Immunization with a recombinant BibA surface protein confers immunity and protects mice against group B Streptococcus (GBS) vaginal colonization

Streptococcus agalactiae or group B Streptococcus (GBS) is a Gram-positive bacterium divided into ten distinct serotypes that colonizes the vaginal and rectal tracts of approximately 30% of women worldwide. GBS is the leading cause of invasive infection in newborns, causing sepsis, pneumoniae and meningitis. The main strategy to prevent GSB infection in newborns includes the use of intrapartum antibiotic therapy, which does not prevent late-onset diseases and may select resistant bacterial strains. We still do not have a vaccine formulation specific for this pathogen approved for human use. Conserved surface proteins are potential antigens that could be targets for recognition by antibodies and activation of cell opsonization. We used a serotype V GBS (GBS-V)-derived recombinant surface protein, rBibA, and evaluated the potential protective role of the induced antigen-specific antibodies after parenteral or mucosal immunizations in C57BL/6 mice. In vitro and in vivo assays demonstrated that vaccine formulations containing BibA combined with different adjuvants induced serum IgG and/or secreted IgA antibodies, leading to enhanced opsonophagocytosis of GBS-V cells and reduced invasion of epithelial cells. One BibA-based vaccine formulation adjuvanted with a nontoxic derivative of the heat-labile toxin produced by enterotoxigenic Escherichia coli (ETEC) strains was capable of inducing protection against vaginal colonization and lethal parenteral challenge with GBS-V. Serum collected from vaccinated mice conferred passive protection against vaginal colonization in naïve mice challenged with GBS-V. Taken together, the present data demonstrate that the BibA protein is a promising antigen for development of a vaccine to protect against GBS infection.

Staphylococcus aureus nasal and intestinal carriage by free-ranging red deer: evidence of human, domestic and wild animal lineages

Purpose: Habitat loss and fragmentation, caused by human activities, can trigger interactions among humans, domestic and wild animals, creating opportunities for the transmission of infectious agents. In Stelvio National Park (SNP, Central Italian Alps, Lombardy sector), the increase of the red deer population, associated with an intense browsing impact and cross transmission of pathogens with livestock, imposed the activation of a culling plan. Moreover, culled deer from SNP entered the human food chain with 37.3 tons of meat in 2011-16. This study evaluates the prevalence and genetic characteristics of Staphylococcus aureus isolates from nasal cavities and rectal tracts in red deer culled in SNP in January and February 2017. Methods & Materials: Both nasal swabs and feces or rectal swabs, collected from 75 red deer, were incubated in Mueller-Hinton broth with 6.5% NaCl and seeded on Baird-Parker with rabbit plasma fibrinogen. S. aureus was confirmed by amplification of the nuc gene. One isolate per collection site from simultaneous nasal and intestinal carriers was characterized using a S. aureus-specific DNA microarray, which detects over 333 genes and alleles. Results: A S. aureus prevalence of 90.67% (95% CI: 81.97–95.41) and 26.67% (95% CI:17.98–37.63) was detected in nasal and intestinal site respectively. All intestinal carriers (n = 20) had also a nasal colonization. Clonal complex (CC) 425, known to be widespread in ruminants, was the most prevalent lineage (68.29%). Notably, 67.86% of CC425 strains carried the leukocidin genes lukM/lukF-P83, which are not typical of this lineage. Further lineages identified included some that are also known to infect humans and/or livestock (CC7, CC9, CC350, CC707) and some (CC2328, CC2671) that are so rare that no epidemiological information is available. All isolates were negative for methicillin resistance genes, but human lineages harbored penicillin resistance genes and one isolate also ermA gene. Conclusion: A high prevalence of S. aureus carriers has been observed among free-ranging red deer during the cold season in the Alps. Red deer S. aureus population mainly comprised lineages known to occur in domestic and wild ruminants. Sporadic overlaps with human and other animal lineages were observed.

Evaluation of the effect of photodynamic therapy with hematoporphyrin monomethyl ether on VX2 tumors implanted in the rectal submucosa of rabbits

We evaluated the influence of the photodynamic therapy (PDT) dosage with hematoporphyrin monomethyl ether (HMME) on its therapeutic efficacy in a rabbit model of in situ rectal cancer. VX2 cells were injected into the rectal submucosa of a rabbit to establish a carcinoma model in situ. After 10–14days, the tumors were treated with PDT at three dosages: a low dose (5mg/kg HMME; 60J/cm2 laser power), intermediate dose (5mg/kg; 240J/cm2), and high dose (10mg/kg; 360J/cm2). Tumor growth, animal survival, histopathological changes, general conditions, and adverse reactions were analyzed. PDT showed inhibitive effects on rectal cancer in all PDT groups, and the efficacy was correlated with the PDT dosage. The high dose PDT group had the best efficacy with a remarkable response rate of 20% and slight response rate of 80%, but it also had the highest death rate of 80% at day 7. The intermediate dose PDT group had a total response rate of 80% (60% remarkable plus 20% slight) and a 40% death rate. Comparably, the low dose PDT group had a 40% slight response rate and 60% death rate. Therefore, the intermediate dose of PDT was considered to be optimal among the three groups. Based on endoscopy findings, we also found that high dose PDT presented more side effects including inflammation, intestinal obstruction, rectal dysfunction, and death when PDT was performed on tumors inside the rectal tract. Our results indicate that a moderate PDT dose is more appropriate to treat tumors located in tract or cavity tissues.

Antiretroviral Pharmacology in Mucosal Tissues

Strategies to prevent HIV infection using preexposure prophylaxis are required to curtail the HIV pandemic. The mucosal tissues of the genital and rectal tracts play a critical role in HIV acquisition, but antiretroviral (ARV) disposition and correlates of efficacy within these tissues are not well understood. Preclinical and clinical strategies to describe ARV pharmacokinetic-pharmacodynamic relationships within mucosal tissues are currently being investigated. In this review, we summarize the physicochemical and biologic factors influencing ARV tissue exposure. Furthermore, we discuss the necessary steps to generate relevant pharmacokinetic-pharmacodynamic data and the challenges associated with this process. Finally, we suggest how preclinical and clinical data might be practically translated into optimal preexposure prophylaxis dosing strategies for clinical trials testing using mathematical modeling and simulation.

A proteomic profile of washing fluid from the colorectal tract to search for potential biomarkers of colon cancer

Washing fluid (WF) from the colon rectal tract after surgical resection might represent a first step in obtaining a mixture of proteins derived from the secretion of tumoral epithelial cells potentially involved in the pathological progression of tissue. In this study, we performed a proteomic analysis of colorectal WF to search for potential biomarkers of colon cancer. The outcome of this approach might open the possibility of using WF to screen for the precancerous and early stages of colorectal cancer (CRC). Samples of WFs were obtained during surgery from 35 patients submitted to colon resection for suspicious adenocarcinoma or carcinoma, while the respective controls were obtained by washing the healthy sections. WFs were immediately centrifuged, concentrated and trichloroacetic acid (TCA) was added to obtain protein pellets. After two-dimensional gel electrophoresis (2DE), the protein patterns of malignant samples were compared with respective normal samples. Forty-one protein spots were found to be differentially expressed exhibiting ≥2 fold-change of mean value spot intensities. After mass spectrometry, these protein spots collapsed into 38 different proteins. Interestingly, 19 of the differentially expressed proteins identified in the study corresponded to those suggested as being potential biomarkers of CRC. In accordance with the literature, these proteins showed the same direction of change (up or down for all proteins). Our results suggest that WF has the potential of being a method for the exploration of clinical samples for biomarker and drug target discovery.

Fine needle aspiration cytology of primary or recurrent chordomas

Chordomas are infrequent, recurrent neoplasms, which represent 1–4% of all primary bone tumours. They originate in notochord remnants located in vertebral bodies and intervertebral discs. Chordomas can also originate from ectopic remains of the notochord in paravertebral soft tissue or at the end of the neural axis. They are slow growing tumours, frequently located in the spine, sacral or spheno-occipital regions. The cervical, thoracic and lumbar locations are less frequent. Sacral chordomas are usually felt at rectal tract exploration, but not detected at lumbar examination, unless the tumour is large and with soft tissue involvement. Chordomas are not gender specific and occur mostly in the fourth or fifth decades of life, although some cases have been described in patients as young as 30 years of age. Generally, chordomas are considered low or intermediate grade lesions. On X-ray, chordomas are osteolytic lesions that can be calcified and involve bones and soft tissue. The diagnosis is suspected on imaging and then confirmed on histology. There are few cases published regarding the diagnosis of chordomas by fine needle aspiration (FNA) cytology. This technique has been described mainly for the differential diagnosis with other tumours and to confirm recurrences. We present two cases of chordomas, whose diagnosis was obtained first by cytology with subsequent histological confirmation. Case histories

Recombinant poxviruses as mucosal vaccine vectors

The majority of infections initiate their departure from a mucosal surface, such as Human immunodeficiency virus (HIV), a sexually transmitted virus. Therefore, the induction of mucosal immunity is a high priority in the development of vaccines against mucosal pathogens. The selection of an appropriate antigen delivery system is necessary to induce an efficient mucosal immune response. Poxvirus vectors have been the most intensively studied live recombinant vector, and numerous studies have demonstrated their ability to induce mucosal immune responses against foreign expressed antigens. Previous studies have demonstrated that recombinants based on the attenuated modified vaccinia virus Ankara (MVA) vector were effective in inducing protective responses against different respiratory viruses, such as influenza and respiratory syncytial virus, following immunization via mucosal routes. Recent studies performed in the murine and macaque models have shown that recombinant MVA (rMVA) does not only stimulate HIV-specific immunity in the genital and rectal tracts following mucosal delivery, but can also control simian/human immunodeficiency viraemia and disease progression. In addition, a prime-boost vaccination approach against tuberculosis emphasized the importance of the intranasal rMVA antigen delivery to induce protective immunity against Mycobacterium tuberculosis. The aim of this review is to summarize the studies employing recombinant poxviruses, specifically rMVA as a mucosal delivery vector. The results demonstrate that rMVAs can activate specific immune responses at mucosal surfaces, and encourage further studies to characterize and improve the MVA mucosal immunogenicity of poxvirus vectors.

Prime-boost immunization schedules based on influenza virus and vaccinia virus vectors potentiate cellular immune responses against human immunodeficiency virus Env protein systemically and in the genitorectal draining lymph nodes.

Vaccines that elicit systemic and mucosal immune responses should be the choice to control human immunodeficiency virus (HIV) infections. We have previously shown that prime-boost immunizations with influenza virus Env and vaccinia virus (VV) WR Env recombinants induced an enhanced systemic CD8(+) T-cell response against HIV-1 Env antigen. In this report, we analyzed in BALB/c mice after priming with influenza virus Env the ability of two VV recombinants expressing HIV-1 Env B (VV WR Env and the highly attenuated modified VV Ankara [MVA] Env) to boost cellular immune responses in the spleen and in the lymph nodes draining the genital and rectal tracts. Groups of mice were primed by the intranasal route with 10(4) PFU of influenza virus Env and boosted 14 days later by the intraperitoneal or intranasal route with 10(7) PFU of MVA Env or VV WR Env, while the control group received two immunizations with influenza virus Env. We found that the combined immunization (Flu/VV) increased more than 60 times the number of gamma interferon-specific CD8(+) T cells compared to the Flu/Flu scheme. Significantly, boosting with MVA Env by the intraperitoneal route induced a response 1.25 or 2.5 times (spleen or genital lymph nodes) higher with respect to that found after the boost with VV WR Env. Mice with an enhanced CD8(+) T-cell response also had an increased Th1/Th2 ratio, evaluated by the cytokine pattern secreted following in vitro restimulation with gp160 protein and by the specific immunoglobulin G2a (IgG2a)/IgG1 ratio in serum. By the intranasal route recombinant WR Env booster gave a more efficient immune response (10 and 1.3 times in spleen and genital lymph nodes, respectively) than recombinant MVA Env. However, the scheme influenza virus Env/MVA Env increased four times the response in the spleen, giving a low but significant response in the genital lymph nodes compared with a single intranasal immunization with MVA Env. These results demonstrate that the combination Flu/MVA in prime-booster immunization regimens is an effective vaccination approach to generate cellular immune responses to HIV antigens at sites critical for protective responses.

Systemic and mucosal immune responses in mice after mucosal immunization with group B streptococcus type III capsular polysaccharide-cholera toxin B subunit conjugate vaccine.

Group B streptococci (GBS) colonize the female genital and rectal tracts and can cause invasive infection in susceptible newborns. An optimally effective GBS vaccine should induce mucosal and systemic immunity. In this study, we investigate the local and systemic immune responses to GBS type III capsular polysaccharide (CPS) after mucosal vaccination of mice via intranasal, peroral, rectal, and vaginal routes, with GBS type III CPS conjugated with recombinant cholera toxin B subunit (GBS III CPS-rCTB). Cholera toxin (CT) was added as an adjuvant. Immunoglobulin G (IgG) and IgA antibodies to the CPS were tested in serum, lungs, and intestinal, rectal, and vaginal extracts by enzyme-linked immunosorbent assay. The conjugated CPS administered by intranasal, peroral, rectal, and vaginal routes was much more effective at inducing both mucosal and systemic antibody responses to GBS III CPS than was unconjugated CPS. The CPS-specific immune responses in various organs were dependent on the route of immunization. Generally, the highest levels of IgA and IgG were generated in the regions or sites of the conjugate exposure. Thus, intranasal vaccination elicited the highest anti-CPS IgA and IgG antibody levels in the lungs, whereas peroral administration in the intestinal site and vaginal vaccination elicited the highest antibody levels in the vagina. Rectal vaccination was superior to the other routes in inducing high antibody levels in the rectum. The four routes of mucosal vaccination also induced distant antibody responses to CPS. Rectal vaccination induced high specific IgA levels in the vagina and intestine, and oral administration induced high specific IgA levels in the lungs and rectum. All four routes of vaccination with the conjugate elicited similarly high levels of anti-CPS IgG in serum. Intranasal vaccination with different doses of the conjugate (10, 30, and 80 microg of CPS) did not have a significant influence on the anti-CPS specific antibody responses. Intranasal immunization induced better antibody responses when one dose of the conjugate was divided and given on three consecutive days compared to administration of the full dose on one occasion. In conclusion, rectal and vaginal vaccination may be the best way of stimulating anti-CPS immune responses in the rectal and vaginal tracts, while high levels of anti-CPS antibodies in the lungs can be achieved after intranasal administration. The vaccination regimen thus might influence the mucosal immune response to CPS. This conjugate may serve as an effective mucosal vaccine for preventing mucosal colonization and invasive infection caused by GBS.

Intranasal Immunization with Plasmid DNA-Lipid Complexes Elicits Mucosal Immunity in the Female Genital and Rectal Tracts

The development of vaccines against pathogens transmitted across the genito-rectal mucosa that effectively stimulate both secretory IgA Abs and cytotoxic T lymphocytes in the genital tract and CTL in the draining lymph nodes (LN) has proven a major challenge. Here we report a novel, noninvasive approach of genetic vaccination via the intranasal route. Such vaccination elicits immune responses in the genital and rectal mucosa, draining LNs, and central lymphoid system. Intranasal immunization with plasmid DNA-lipid complexes encoding the model Ag firefly luciferase resulted in dissemination of the DNA and the encoded transcript throughout the respiratory and gastrointestinal tracts, draining LNs, and spleen. Complexing the plasmid DNA with the lipid DMRIE/DOPE enhanced expression of the encoded protein in the respiratory tract, increased specific secretory IgA Ab in the vaginal and rectal tracts, and increased the circulating levels of specific IgA and IgG. In addition, intranasal DNA immunization resulted in generation of Ag-specific CTL that were localized in the genital and cervical LNs and spleen. These results suggest that intranasal immunization with plasmid DNA-lipid complexes may represent a generic immunization strategy against pathogens transmitted across the genito-rectal and other mucosal surfaces.

In vitro activity of antimycotic agents determined by E-test method against vaginal Candida species.

Vaginal candidiasis continues to be a common cause of vaginal discharge, pruritus and other local complaints in women worldwide. Although numerous antimycotic agents are available for the treatment of yeast vaginitis there is little comparative data on the in vitro activity of these drugs. The objectives of this study were to isolate and identify the Candida species in the vagina and anus of patients treated in a gynaecology clinic, as well as determine the susceptibility to azolic compounds measured by the E-test method. Vaginal and rectal swabs were collected from 80 adult non-pregnant patients, seen at a gynaecological clinic, aged 18-59 years, with sexual activity, with and without vaginitis. The swabs were processed by methods routinely used for the detection of pathogenic yeasts. The susceptibility of the isolates to fluconazole, ketoconazole and itraconazole, was measured by the agar diffusion method (E-test), using RPM1 1,640 medium with 2% glucose and phosphate buffer. Candida species (33) strains were isolated from 17 patients at similar proportions from both anatomical sites, and 12 patients harboured 24 strains of C. albicans in the vaginal and rectal tracts. Twenty one percent of the strains of C. albicans were resistant to ketoconazole, 54% were resistant to itraconazole and 0% were resistant to fluconazole. The sensitivity of strains isolated from the two sites were similar, indicating that these are strains of the same phenotype.