Changes in neuromuscular structure and functions of human colon during ageing are region-dependent.

OBJECTIVE: To determine if human colonic neuromuscular functions decline with increasing age. DESIGN: Looking for non-specific changes in neuromuscular function, a standard burst of electrical field stimulation (EFS) was used to evoke neuronally mediated (cholinergic/nitrergic) contractions/relaxations in ex vivomuscle strips of human ascending and descending colon, aged 35-91 years (macroscopically normal tissue; 239 patients undergoing cancer resection). Then, to understand mechanisms of change, numbers and phenotype of myenteric neurons (30 306 neurons stained with different markers), densities of intramuscular nerve fibres (51 patients in total) and pathways involved in functional changes were systematically investigated (by immunohistochemistry and use of pharmacological tools) in elderly (≥70 years) and adult (35-60 years) groups. RESULTS: With increasing age, EFS was more likely to evoke muscle relaxation in ascending colon instead of contraction (linear regression: n=109, slope 0.49%±0.21%/year, 95% CI), generally uninfluenced by comorbidity or use of medications. Similar changes were absent in descending colon. In the elderly, overall numbers of myenteric and neuronal nitric oxide synthase-immunoreactive neurons and intramuscular nerve densities were unchanged in ascending and descending colon, compared with adults. In elderly ascending, not descending, colon numbers of cell bodies exhibiting choline acetyltransferase immunoreactivity increased compared with adults (5.0±0.6 vs 2.4±0.3 neurons/mm myenteric plexus, p=0.04). Cholinergically mediated contractions were smaller in elderly ascending colon compared with adults (2.1±0.4 and 4.1±1.1 g-tension/g-tissue during EFS; n=25/14; p=0.04); there were no changes in nitrergic function or in ability of the muscle to contract/relax. Similar changes were absent in descending colon. CONCLUSION: In ascending not descending colon, ageing impairs cholinergic function.

Ex vivo study of human visceral nociceptors.

OBJECTIVE: The development of effective visceral analgesics free of deleterious gut-specific side effects is a priority. We aimed to develop a reproducible methodology to study visceral nociception in human tissue that could aid future target identification and drug evaluation. DESIGN: Electrophysiological (single unit) responses of visceral afferents to mechanical (von Frey hair (VFH) and stretch) and chemical (bradykinin and ATP) stimuli were examined. Thus, serosal afferents (putative nociceptors) were used to investigate the effect of tegaserod, and transient receptor potential channel, vanilloid 4 (TRPV4) modulation on mechanical responses. RESULTS: Two distinct afferent fibre populations, serosal (n=23) and muscular (n=21), were distinguished based on their differences in sensitivity to VFH probing and tissue stretch. Serosal units displayed sensitivity to key algesic mediators, bradykinin (6/14 units tested) and ATP (4/10), consistent with a role as polymodal nociceptors, while muscular afferents are largely insensitive to bradykinin (0/11) and ATP (1/10). Serosal nociceptor mechanosensitivity was attenuated by tegaserod (-20.8±6.9%, n=6, p<0.05), a treatment for IBS, or application of HC067047 (-34.9±10.0%, n=7, p<0.05), a TRPV4 antagonist, highlighting the utility of the preparation to examine the mechanistic action of existing drugs or novel analgesics. Repeated application of bradykinin or ATP produced consistent afferent responses following desensitisation to the first application, demonstrating their utility as test stimuli to evaluate analgesic activity. CONCLUSIONS: Functionally distinct subpopulations of human visceral afferents can be demonstrated and could provide a platform technology to further study nociception in human tissue.

Fabrication and characterization of alginate-zein core-shell microcapsules for controlled release of buckwheat honey.

In this study, extrusion method was employed to fabricate alginate-zein core-shell microcapsules loaded with buckwheat honey by dropping alginate and buckwheat honey mixture solution into a 70.0 % zein ethanol solution(v/v) containing 5.0 % CaCl2 solution (wt%). The microcapsules were constructed by two parts: 1) the formation of hydrophilic beads through the crosslinking of alginate chains with Ca2+; 2) the introduction of alginate beads into the aqueous zein ethanol solution which decreased the ethanol concentration, prompting the precipitation of zein and the deposition of zein nanoparticles onto the surfaces of alginate beads. Comparing with the alginate beads, the prepared microcapsules not only possessed better water-holding capacity, but also achieved controlled release of buckwheat honey. Importantly, the microcapsules significantly retained the antioxidant activity of the buckwheat honey. Therefore, this innovative method for fabricating alginate-zein core-shell microcapsules can suggest a promising approach to broaden the application of buckwheat honey in the food field.

Analysis of colorectal cancers in British Bangladeshi identifies early onset, frequent mucinous histotype and a high prevalence of RBFOX1 deletion.

BACKGROUND: Prevalence of colorectal cancer (CRC) in the British Bangladeshi population (BAN) is low compared to British Caucasians (CAU). Genetic background may influence mutations and disease features. METHODS: We characterized the clinicopathological features of BAN CRCs and interrogated their genomes using mutation profiling and high-density single nucleotide polymorphism (SNP) arrays and compared findings to CAU CRCs. RESULTS: Age of onset of BAN CRC was significantly lower than for CAU patients (p=3.0 x 10-5) and this difference was not due to Lynch syndrome or the polyposis syndromes. KRAS mutations in BAN microsatellite stable (MSS) CRCs were comparatively rare (5.4%) compared to CAU MSS CRCs (25%; p=0.04), which correlates with the high percentage of mucinous histotype observed (31%) in the BAN samples. No BRAF mutations was seen in our BAN MSS CRCs (CAU CRCs, 12%; p=0.08). Array data revealed similar patterns of gains (chromosome 7 and 8q), losses (8p, 17p and 18q) and LOH (4q, 17p and 18q) in BAN and CAU CRCs. A small deletion on chromosome 16p13.2 involving the alternative splicing factor RBFOX1 only was found in significantly more BAN (50%) than CAU CRCs (15%) cases (p=0.04). Focal deletions targeting the 5' end of the gene were also identified. Novel RBFOX1 mutations were found in CRC cell lines and tumours; mRNA and protein expression was reduced in tumours. CONCLUSIONS: KRAS mutations were rare in BAN MSS CRC and a mucinous histotype common. Loss of RBFOX1 may explain the anomalous splicing activity associated with CRC.

Heavy metals contamination in shrimp and crab from southwest regions in Bangladesh: Possible health risk assessment.

Shrimp and Crab, important sources of protein, are currently being adversely affected by the rising industrialization, which has led to higher levels of heavy metals. The goal of this study was to evaluate the health risks of contamination associated with nine heavy metals (Cd, Pb, Cu, Cr, Zn, Ni, As, Al, and Fe) in two species of shrimp (Macrobrachium rosenbergii and Metapenaeus monoceros) and one species of crab (Scylla serrata) that were collected from the Khulna, Satkhira, and Bagerhat areas of Bangladesh. Inductively coupled plasma-optical emission spectrometry (ICP-OES) was used for the study. The results showed that all metal concentrations in shrimp and crab samples were below the recommended level, indicating that ingestion of these foods would not pose any substantial health risks to individuals. To evaluate the non-carcinogenic health risks, the target hazard quotient (THQ) and hazard index (HI) were determined, and the target cancer risk (TR) was utilized to evaluate the carcinogenic health risks. From the health point of view, this study showed that crustaceans obtained from the study sites were non - toxic (THQ and HI < 1), and long-term, continuous intake is unlikely to pose any significant health hazards (TR = 10-7-10-5) from either carcinogenic or non-carcinogenic effects.

State-of-the-art biosynthesis of tin oxide nanoparticles by chemical precipitation method towards photocatalytic application.

Tin oxide (SnO2) with versatile properties is of substantial standing for practical application, and improved features of the material are demonstrated in the current issue through the integration of nanotechnology with bio-resources leading to what is termed as biosynthesis of SnO2 nanoparticles (NPs). This review reveals the recent advances in biosynthesis of SnO2 NPs by chemical precipitation method focused on distinct methodologies, characterization, and reaction mechanism along with a photocatalytic application for dye degradation. According to available literature reviews, numerous bio-based precursors selectively extracted from biological substrates have effectively been applied as capping or reducing agents to achieve the metal oxide NPs. The major precursor obtained from the aqueous extract of root barks of Catunaregam spinosa is found to be 7-hydroxy-6-methoxy-2H-chromen-2-one that has been proposed as a model compound for the reduction of metal ions into nanoparticles due to having highly active functional groups, being abundant in plants (67.475 wt%), easy to extract, and eco benign. In addition, the photocatalytic activity of SnO2 NPs for the degradation of organic dyes, pharmaceuticals, and agricultural contaminants has been discussed in the context of a promising bio-reduction mechanism of the synthesis. The final properties are supposed to depend exclusively upon a number of factors, e.g., particle size (< 50 nm), bandgap (< 3.6 eV), crystal defects, and catalysts dosage. With this contribution, it has been perceived not only to provide an overview of recent advances in the biosynthesis of SnO2 NPs but also to indicate the main issues in need aiming to show vision towards innovative outcomes.

A Comprehensive Review on Bio-Preservation of Bread: An Approach to Adopt Wholesome Strategies.

Bread is a food that is commonly recognized as a very convenient type of food, but it is also easily prone to microbial attack. As a result of bread spoilage, a significant economic loss occurs to both consumers and producers. For years, the bakery industry has sought to identify treatments that make bread safe and with an extended shelf-life to address this economic and safety concern, including replacing harmful chemical preservatives. New frontiers, on the other hand, have recently been explored. Alternative methods of bread preservation, such as microbial fermentation, utilization of plant and animal derivatives, nanofibers, and other innovative technologies, have yielded promising results. This review summarizes numerous research findings regarding the bio-preservation of bread and suggests potential applications of these techniques. Among these techniques, microbial fermentation using lactic acid bacteria strains and yeast has drawn significant interest nowadays because of their outstanding antifungal activity and shelf-life extending capacity. For example, bread slices with Lactobacillus plantarum LB1 and Lactobacillus rossiae LB5 inhibited fungal development for up to 21 days with the lowest contamination score. Moreover, various essential oils and plant extracts, such as lemongrass oil and garlic extracts, demonstrated promising results in reducing fungal growth on bread and other bakery products. In addition, different emerging bio-preservation strategies such as the utilization of whey, nanofibers, active packaging, and modified atmospheric packaging have gained considerable interest in recent days.

Critical Assessment of Mycotoxins in Beverages and Their Control Measures.

Mycotoxins are secondary metabolites of filamentous fungi that contaminate food products such as fruits, vegetables, cereals, beverages, and other agricultural commodities. Their occurrence in the food chain, especially in beverages, can pose a serious risk to human health, due to their toxicity, even at low concentrations. Mycotoxins, such as aflatoxins (AFs), ochratoxin A (OTA), patulin (PAT), fumonisins (FBs), trichothecenes (TCs), zearalenone (ZEN), and the alternaria toxins including alternariol, altenuene, and alternariol methyl ether have largely been identified in fruits and their derived products, such as beverages and drinks. The presence of mycotoxins in beverages is of high concern in some cases due to their levels being higher than the limits set by regulations. This review aims to summarize the toxicity of the major mycotoxins that occur in beverages, the methods available for their detection and quantification, and the strategies for their control. In addition, some novel techniques for controlling mycotoxins in the postharvest stage are highlighted.

Nutritional characterization of freshwater mud eel (Monopterus cuchia) muscle cooked by different thermal processes.

This paper reports the effects of four popular cooking methods viz. grilling, boiling, frying, and microwaving on the proximate and nutritional compositions of freshwater mud eel (FWME) muscle. The moisture content of raw FWME muscle was 74.45%, which was similar in boiled products but lower in grilled, fried, and microwaved products (p ≤ .05). The protein content in raw and cooked FWME muscles varied between 14.49% and 21.28%. There were found 20 different fatty acids in FWME muscle of which palmitic acid was the most abundant one with an amount of 26.51%-29.70% in raw and cooked FWME muscles. FWME muscle contained a substantial amount of ω-3 polyunsaturated fatty acids, ranging from 7.54% to 13.7%. However, the thermal effects during cooking decreased the ω-3 polyunsaturated fatty acid contents. There were seven essential and eight nonessential amino acids available in FWME muscle; among the essential amino acids, lysine content was the highest. Raw and cooked FWME were very rich in calcium, between 794.52 mg/100 g and 883.24 mg/100 g muscle. Among the studied heavy metals, Pb content was the highest. However, all the heavy metal contents were within acceptable limits determined by health risk assessment, that is, target hazard quotient and target cancer risk.

The impact of medical student surgical conferences.

BACKGROUND: Recent reports have shown that in the UK there has been a decrease in interest towards pursuing a surgical career, whereas early exposure to surgery may increase interest. We aimed to assess the effect of a 1-day medical school surgical conference in encouraging attendees to pursue a surgical career. METHODS: All attendees of the 2014 Barts and The London National Undergraduate Surgical conference were invited to participate in an online survey. Data were collected and analysed to evaluate students' reasons for attending the conference, and attitudes to careers in surgery before and after the conference. RESULTS: Of 229 students, 130 (56.8%) completed the survey. Year-12 high-school and preclinical medical students showed a statistically significant difference in interest in pursuing a surgical career after the conference than before it (p = 0.0002 and p = 0.0027, respectively), but clinical medical students demonstrated no such change. CONCLUSION: Attendance at surgical conferences designed for medical students can significantly increase the desire of high-school students and preclinical medical students to pursue a surgical career, but may not have the same effect for clinical medical students. Surgical conferences may act as an effective means of recruiting students towards choosing a surgical career for a certain subset of students.

Relative telomere lengths in tumor and normal mucosa are related to disease progression and chromosome instability profiles in colorectal cancer.

Telomeric dysfunction is linked to colorectal cancer (CRC) initiation. However, the relationship of normal tissue and tumor telomere lengths with CRC progression, molecular features and prognosis is unclear. Here, we measured relative telomere length (RTL) by real-time quantitative PCR in 90 adenomas (aRTL), 419 stage I-IV CRCs (cRTL) and adjacent normal mucosa (nRTL). Age-adjusted RTL was analyzed against germline variants in telomere biology genes, chromosome instability (CIN), microsatellite instability (MSI), CpG island methylator phenotype (CIMP), TP53, KRAS, BRAF mutations and clinical outcomes. In 509 adenoma or CRC patients, nRTL decreased with advancing age. Female gender, proximal location and the TERT rs2736100 G allele were independently associated with longer age-adjusted nRTL. Adenomas and carcinomas exhibited telomere shortening in 79% and 67% and lengthening in 7% and 15% of cases. Age-adjusted nRTL and cRTL were independently associated with tumor stage, decreasing from adenoma to stage III and leveling out or increasing from stage III to IV, respectively. Cancer MSI, CIMP, TP53, KRAS and BRAF status were not related to nRTL or cRTL. Near-tetraploid CRCs exhibited significantly longer cRTLs than CIN- and aneuploidy CRCs, while cRTL was significantly shorter in CRCs with larger numbers of chromosome breaks. Age-adjusted nRTL, cRTL or cRTL:nRTL ratios were not associated with disease-free or overall survival in stage II/III CRC. Taken together, our data show that both normal mucosa and tumor RTL are independently associated with CRC progression, and highlight divergent associations of CRC telomere length with tumor CIN profiles.

Intraoperative monitoring of intestinal viability: Evaluation of a new combined sensor.

A dual wavelength photoplethysmography (PPG) and laser Doppler flowmetry (LDF) sensor was developed to investigate the suitability of these techniques for monitoring bowel viability intraoperatively. Clinical measurements were obtained from thirty patients undergoing bowel surgery. Three measurements were performed at different stages of the operation. The amplitude of infrared PPG decreased from the baseline measurement to the pre-anastomosis measurement by 36% and LDF flux decreased by 21% for the same measurements. An increase of 33% in amplitude for infrared PPG was observed from the pre-anastomotic to post-anastomosis measurement; the equivalent increase was not seen for LDF flux. The results revealed that the sensor could potentially indicate changes in perfusion and blood flow at critical phases of surgery, thereby assisting in the early detection of inadequate blood supply in bowel tissue. The results also suggest that laser Doppler is more sensitive to movement artefact compared to PPG.

Colorectal cancers with microsatellite instability display mRNA expression signatures characteristic of increased immunogenicity.

BACKGROUND: Colorectal cancers displaying high-degree microsatellite instability (MSI-H) have an improved prognosis compared to microsatellite stable (MSS) cancers. The observation of pronounced lymphocytic infiltrates suggests that MSI-H cancers are inherently more immunogenic. We aimed to compare the gene expression profiles of MSI-H and MSS cancers to provide evidence for an activated immune response in the former. RESULTS: We analysed tissue from 133 colorectal cancer patients with full consent and Local Ethics Committee approval. Genomic DNA was analysed for microsatellite instability in BAT-26. High-quality RNA was used for microarray analysis on the Affymetrix HG-U133A chip. Data was analysed on GeneSpring software version 6.0. Confirmatory real-time RT-PCR was performed on 28 MSI-H and 26 MSS cancers. A comparison of 29 MSI-H and 104 MSS cancers identified 2070 genes that were differentially expressed between the two groups [P < 0.005]. Significantly, many key immunomodulatory genes were up-regulated in MSI-H cancers. These included antigen chaperone molecules (HSP-70, HSP-110, Calreticulin, gp96), pro-inflammatory cytokines (Interleukin (IL)-18, IL-15, IL-8, IL-24, IL-7) and cytotoxic mediators (Granulysin, Granzyme A). Quantitative RT-PCR confirmed up-regulation of HSP-70 [P = 0.016], HSP-110 [P = 0.002], IL-18 [P = 0.004], IL-8 [0.002] and Granulysin [P < 0.0001]. CONCLUSIONS: The upregulation of a large number of genes implicated in immune response supports the theory that MSI-H cancers are immunogenic. The novel observation of Heat Shock Protein up-regulation in MSI-H cancer is highly significant in light of the recognised roles of these proteins in innate and antigen-specific immunogenicity. Increased mRNA levels of pro-inflammatory cytokines and cytotoxic mediators also indicate an activated anti-tumour immune response.

Instructional improvement: how to overcome the barriers?

This is no easy way to improve teaching. Not one or even two activities are right or best for all instructions. The improvement of instruction requires a multi directional approach. When faculty members, departmental heads or interested administrator come across an idea they find appealing, they often tend to be converted and start proselytizing. The way to better teaching lies in review of videotaped teaching samples or diagnostic student evaluation with consultation, or seminars on course design, or monitoring with master teachers, peer assessment or whatever the favored strategy happens to be. All of these are possible ways to better teaching, made right or best only after they have been carefully matched with their instructional needs of the teacher, course content & instructional setting.

Quantification of cytokeratin 20, carcinoembryonic antigen and guanylyl cyclase C mRNA levels in lymph nodes may not predict treatment failure in colorectal cancer patients.

Conventional histopathologic staging of primary colorectal cancers does not allow accurate prognostic stratification within a given tumour stage. Therefore, PCR-based assays are increasingly used to try to predict more accurately the likelihood of disease progression for the individual patient. Real-time reverse transcription PCR (RT-PCR) assays were used to detect and quantitate cytokeratin 20 (ck20), carcinoembryonic antigen (CEA) and guanylyl cyclase C (GCC) mRNA in 149 lymph nodes (LN) from 17 patients with benign disease and 302 LN from 42 patients with colorectal cancer who had curative (R0) resections. None of the markers were specific, with ck20, CEA and GCC mRNA detected in 47%, 89% and 13% of 149 LN, respectively, from patients with benign disease. The sensitivity of all 3 markers was very high, with mRNA detected in 93%, 100% and 97% of 30 histologically involved LN, respectively. There was significant overlap in the mRNA levels of all 3 markers between histologically involved and uninvolved LN. There was no association between mRNA levels and distant recurrence (median follow-up: 3.94 years, range 3.35-5.12). We conclude that the use of molecular techniques to detect occult disease in LN may suffer from the same limitations as conventional methods. Instead, accurate prognostic stratification requires careful assessment of the likely metastatic potential of the primary cancer.

Differential expression patterns of the insulin-like growth factor 2 gene in human colorectal cancer.

Tumour development and metastasis are associated with altered gene expression profiles. The aim of this study was to identify the transcriptional differences in normal, tumour and metastatic tissue. We used oligonucleotide arrays to identify differential expression patterns of insulin-like growth factor 2 (IGF 2) between 139 primary colorectal tumour specimens and 42 tumour-adjacent mucosa specimens from colorectal cancer (CRC) patients. The expression levels of the IGF 2 gene were significantly increased in primary tumours compared with adjacent mucosae. This was concordant with our real-time RT-PCR quantification of 48 matched tumour mucosa samples. IGF 2 expression levels were also measured by RT-PCR quantitative analysis in 18 liver metastases and 10 normal tissues from patients without cancer. The mRNA levels were significantly under-expressed in liver metastases compared with either colorectal tumours or adjacent normal mucosae. The non- malignant normal tissue expressed significantly lower IGF 2 levels than adjacent normal tissue, and this was not due to a field effect originating from the tumour. In addition, our microarray data demonstrated that IGF 2 expression was down-regulated in sporadic microsatellite instability (MSI-H) CRC and parallels under-expression of hMLH1 and IGF 2 receptor genes in these patients. We conclude that IGF 2 plays an important role in CRC development. Also, individuals with loss of genomic imprinting (LOI) causing over-expression of IGF 2 may be at greater risk of developing CRC. However, this LOI may be reversed in MSI-H patients.