Search

Background: Adenoid cystic carcinoma is a rare subtype of triple‐negative breast carcinoma. These low‐grade tumours, which are treated by simple mastectomy and have an excellent prognosis compared to other triple‐negative breast carcinomas. Solid‐variant adenoid cystic carcinomas have basaloid features and are difficult to distinguish morphologically from other triple‐negative breast cancers. Breast adenoid cystic carcinoma exhibits MYB protein overexpression, which can be detected by immunohistochemistry (IHC). Aim: We compared the IHC expression of MYB in solid‐variant adenoid cystic carcinoma with that in other triple‐negative breast cancers. Methods: We conducted IHC staining of 210 samples of triple‐negative breast cancers, including solid‐variant adenoid cystic carcinoma (n = 17), metaplastic breast carcinoma (n = 44), basaloid triple‐negative breast cancer (n = 21), and other triple‐negative invasive ductal carcinoma (n = 128). We classified nuclear staining of MYB as diffuse/strong (3+), focal moderate (2+), focal weak (1+), or none (0). Results: All 17 solid/basaloid adenoid cystic carcinoma cases exhibited 3+ MYB expression. Of the 21 solid/basaloid triple‐negative breast cancers, one (5%) had 2+ expression, seven (33%) 1+ expression, and 13 (62%) 0 expression. Of the 44 metaplastic carcinoma cases, 39 cases (89%) had no (0) staining, and the other five cases had focal weak (1+) or moderate (2+) staining. Among the 128 triple‐negative invasive ductal carcinoma cases, 92 cases (72%) had no (0) staining, 36 cases (28%) exhibited focal weak (1+) or moderate (2+) staining. Conclusions: Our study revealed diffuse/strong MYB staining (3+) only in solid/basaloid adenoid cystic carcinomas. Thus, we recommend routine MYB IHC staining in triple‐negative breast carcinoma with solid/basaloid morphology to improve diagnostic accuracy. [ABSTRACT FROM AUTHOR]

Simple Summary: Invasive lobular carcinoma (ILC), the second most common type of breast cancer, is a distinct entity. Despite their unique biology and clinical course, lobular carcinomas have been researched in the broad category of hormone receptor positive breast cancers and these too have been predominated by invasive ductal cancers. With advancements in digital techniques, clinical research targeting ILC specifically is imperative in this era of targeted therapy. In this review, we highlighted the most important recent and relevant developments that have happened in the ILC domain, including histology and integrating advances in genomics and the overall translational medicine aspects of ILC. The second most common breast carcinoma, invasive lobular carcinoma, accounts for approximately 15% of tumors of breast origin. Its incidence has increased in recent times due in part to hormone replacement therapy and improvement in diagnostic modalities. Although believed to arise from the same cell type as their ductal counterpart, invasive lobular carcinomas (ILCs) are a distinct entity with different regulating genetic pathways, characteristic histologies, and different biology. The features most unique to lobular carcinomas include loss of E-Cadherin leading to discohesion and formation of a characteristic single file pattern on histology. Because most of these tumors exhibit estrogen receptor positivity and Her2 neu negativity, endocrine therapy has predominated to treat these tumors. However novel treatments like CDK4/6 inhibitors have shown importance and antibody drug conjugates may be instrumental considering newer categories of Her 2 Low breast tumors. In this narrative review, we explore multiple pathological aspects and translational features of this unique entity. In addition, due to advancement in technologies like spatial transcriptomics and other hi-plex technologies, we have tried to enlist upon the characteristics of the tumor microenvironment and the latest associated findings to better understand the new prospective therapeutic options in the current era of personalized treatment. [ABSTRACT FROM AUTHOR]

The present study was aimed to establish the antagonistic effects of Ocimum sanctum L. essential oil (OSEO) on growth and zearalenone (ZEA) production of Fusarium graminearum. GC-MS chemical profiling of OSEO revealed the existence of 43 compounds and the major compound was found to be eugenol (34.7%). DPPH free radical scavenging activity (IC50) of OSEO was determined to be 8.5µg/mL. Minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC) of OSEO on F. graminearum were recorded as 1250 µg/mL and 1800 µg/mL, respectively. Scanning electron microscope observations showed significant micro morphological damage in OSEO exposed mycelia and spores compared to untreated control culture. Quantitative UHPLC studies revealed that OSEO negatively effected the production of ZEA; the concentration of toxin production was observed to be insignificant at 1500 µg/mL concentration of OSEO. On other hand ZEA concentration was quantified as 3.23 µg/mL in OSEO untreated control culture. Reverse transcriptase qPCR analysis of ZEA metabolic pathway genes (PKS4 and PKS13) revealed that increase in OSEO concentration (250 µg/mL to 1500 µg/mL) significantly downregulated the expression of PKS4 and PKS13. These results were in agreement with the artificially contaminated maize grains as well. In conlusion, the antifungal and antimycotoxic effects of OSEO on F. graminearum in the present study reiterated that, the essential oil of O. sanctum could be a promising herbal fungicide in food processing industries as well as grain storage centers.

The enzyme β-galactosidase (β-gal) has extensively used for improvement of lactose intolerance condition. Present study, was designed to assess the potential of β-gal enzyme produced by Lactobacillus fermentum, a kefir isolate, as a biocatalyst for the manufacture of prebiotic galactooligosaccharides (GOS) from lactose. The efficiency of L. fermentum to produce β-gal of 4,254 u/ml was determined by permeabilizing the cells with solvents such as sodium dodecyl sulfate (SDS) and chloroform. Different parameters contributing β-gal production including reaction time, temperature, pH, carbohydrates, and substrate concentration on L. fermentum were studied and optimum β-gal activity was found to be 6,232.13 u/ml. It was observed that different experimental parameters for pH (7.0), temperature (35°C), and carbohydrates (galactose) were statistically significant (p<0.05). L. fermentum was found to produce GOS by transgalactosylation catalysed by β-gal during lactose hydrolysis which yielded di, tri, and tetra oligosaccharides, confirmed by TLC and HPLC. The culture showed β-gal activity, suggesting biotechnological applications and a promising organism for industrial β-gal production. [ABSTRACT FROM AUTHOR]

Biogenic amines (BAs) have attracted increasing attention due to their significance in food safety and quality. The present study aimed at isolation of BAs producing microorganisms from different fermented products, namely kanji, sauerkraut, fish pickle and meat pickle and isolation of BAs degrading bacteria from canine saliva. A total of 63 BAs producing organisms identified among 356 organisms. Initial screening of BAs producing strains was carried out using Moller’s decarboxylase media and 2 BAs degrading organisms identified among 28 organisms. BAs presence is qualitative and quantitative confirmation by TLC and HPLC, respectively, using dansyl derivatized. They were characterized biochemically and identifying using 16S rDNA gene sequencing as Enterobacter hormaechei (18), Enterobacter cloacae (12), Bacillus licheniformis (14), Morganella morganii (11), Lactobacillus sakei (08) and Lactobacillus plantarum (2). Among all isolates, member of the Enterobacteriaceae family yielded the highest amount of putrescine (61.0-244 mg/100 gm), cadaverine (15.8-154 mg/100 gm), histamine (50-210 mg/100 gm) and least amount of tyramine (3-17.2) followed by Bacillus licheniformis which produced putrescine (58.3 mg/100 gm), cadaverine (83.3 mg/100 gm) and histamine (31.7 mg/100 gm). Lactobacillus sakei produced the highest amount of tyramine (21.2 mg/100 gm) and Lactobacillus plantarum from canine saliva degrade all important BAs histamine (100%), tyramine (100%), cadaverine (28%) and putrescine (67.8%). The result of the present study suggests that the BAs producing organisms are frequently present in fermented products, therefore, may pose a potential risk to human health and Lactobacillus plantarum from canine saliva applied as a potential BAs degradation bacterium in foods. [ABSTRACT FROM AUTHOR]

Cytolethal distending toxin ( CDT) producing Campylobacter jejuni species are one of the leading causes of human gastroenteritis worldwide. The main intent of the study was to develop a multiplex PCR assay for the confirmed identification and toxin profiling of C. jejuni. The genes targeted were rpo B as genus specific, hip O for species; cdt A, cdt B, cdt C encoding respective subunit proteins of CDT with Internal Amplification Control ( IAC). To enhance its application as a pre-mixed ready-to-use format, the master mix of developed mPCR was dried by lyophilization and stability was assessed. Thermostabilized reagents showed stability of 1.5 months at room-temperature and upto six months at 4 °C without any loss of functionality. The assay was evaluated on a number of presumptive Campylobacter isolates along with biochemical tests. Results obtained indicated the accurate identification of C. jejuni by developed mPCR format in contrast to misconception associated with biochemical assays. The assay was also tested on spiked samples for its real-time utility. Altogether, the room-temperature storable and ready-to- use mPCR format developed in this study could be preferred for rapid detection and confirmed identification of toxigenic strains of C. jejuni in place of conventional biochemical assays. [ABSTRACT FROM AUTHOR]

In spite of their involvement in foodborne illness, the epidemiological relevance of staphylococcal enterotoxin C (SEC) subtypes is poorly documented may be due to high sequence similarity. Among subtypes, SEC1, SEC2, and SEC3 exhibit more than 97 % homology because of which specific detection tools are seldom available to identify and differentiate them . In this study, a SYBR Green-based RT-PCR followed by melt curve analysis was developed for differentiation of entC1 from entC2/ entC3 using a single primer pair. Nucleotide sequences of all three subtypes were analyzed using Clustal Omega program and the region with significant sequence variation/heterogeneity (where utmost SNPs were closely located and accessible for RT-PCR) was selected for amplification by designing a single primer pair that could amplify all three subtypes. In spite of same amplicon size, entC1 showed distinct melt peak at 76 °C. However, due to high similarity between entC2 and entC3, the developed format was deficient to discriminate between them and both showed melt peak at 82 °C. Reliability of developed RT-PCR was evaluated using various naturally contaminated samples and 91 food and clinical Staphylococcus aureus isolates where satisfactory results were obtained in comparison with commercial immunoassay kit and conventional PCRs using validated primers. To the best of our knowledge, this is the first method being reported to differentiate entC1 from entC2/ entC3 using single primer pair which is unachievable by conventional PCR due to same amplicon size. As benefits, the method is sensitive, rapid, and inexpensive with no requirement of fluorescent probes, multiple primers, and post-PCR procedures. Thus, the assay might find its utility as a detection tool in epidemiological survey of foodborne outbreaks for simultaneous identification and differentiation of entC1 from entC2/ entC3. [ABSTRACT FROM AUTHOR]

Accurate identification of pathogens with biowarfare importance requires detection tools that specifically differentiate them from near-neighbor species. Burkholderia pseudomallei, the causative agent of a fatal disease melioidosis, is one such biothreat agent whose differentiation from its near-neighbor species is always a challenge. This is because of its phenotypic similarity with other Burkholderia species which have a wide spread geographical distribution with shared environmental niches. Melioidosis is a major public health concern in endemic regions including Southeast Asia and northern Australia. In India, the disease is still considered to be emerging. Prevalence surveys of this saprophytic bacterium in environment are under-reported in the country. A major challenge in this case is the specific identification and differentiation of B. pseudomallei from the growing list of species of Burkholderia genus. The objectives of this study included examining the prevalence of B. pseudomallei and near-neighbor species in coastal region of South India and development of a novel detection tool for specific identification and differentiation of Burkholderia species. Briefly, we analyzed soil and water samples collected from Malabar coastal region of Kerala, South India for prevalence of B. pseudomallei. The presumptive Burkholderia isolates were identified using recA PCR assay. The recA PCR assay identified 22 of the total 40 presumptive isolates as Burkholderia strains (22.72% and 77.27% B. pseudomallei and non-pseudomallei Burkholderia respectively). In order to identify each isolate screened, we performed recA and 16S rDNA sequencing. This two genes sequencing revealed that the presumptive isolates included B. pseudomallei, non-pseudomallei Burkholderia as well as non-Burkholderia strains. Furthermore, a gene termed D-beta hydroxybutyrate dehydrogenase (bdha) was studied both in silico and in vitro for accurate detection of Burkholderia genus. The optimized bdha based PCR assay when evaluated on the Burkholderia isolates of this study, it was found to be highly specific (100%) in its detection feature and a clear detection sensitivity of 10 pg/μl of purified gDNA was recorded. Nucleotide sequence variations of bdha among interspecies, as per in silico analysis, ranged from 8 to 29% within the target stretch of 730 bp highlighting the potential utility of bdha sequencing method in specific detection of Burkholderia species. Further, sequencing of the 730 bp bdha PCR amplicon of each Burkholderia strain isolated could differentiate the species and the data was comparable with recA sequence data of the strains. All sequencing results obtained were submitted to NCBI database. Bayesian phylogenetic analysis of bdha in comparison with recA and 16S rDNA showed that the bdha gene provided comparable identification of Burkholderia species. [ABSTRACT FROM AUTHOR]

Lung cancer is a highly aggressive and metastatic disease responsible for approximately 25% of all cancer-related deaths in the United States. Using high-throughput in vitro and in vivo screens, we have previously established Impad1 as a driver of lung cancer invasion and metastasis. Here we elucidate that Impad1 is a direct target of the epithelial microRNAs (miRNAs) miR-200 and miR∼96 and is de-repressed during epithelial-to-mesenchymal transition (EMT); thus, we establish a mode of regulation of the protein. Impad1 modulates Golgi apparatus morphology and vesicular trafficking through its interaction with a trafficking protein, Syt11. These changes in Golgi apparatus dynamics alter the extracellular matrix and the tumor microenvironment (TME) to promote invasion and metastasis. Inhibiting Impad1 or Syt11 disrupts the cancer cell secretome, regulates the TME, and reverses the invasive or metastatic phenotype. This work identifies Impad1 as a regulator of EMT and secretome-mediated changes during lung cancer progression. [Display omitted] • Impad1 is a target of miR-200 and miR∼96 and is de-repressed during EMT • Impad1 regulates Golgi apparatus structure and trafficking to alter exocytosis and TME • Syt11 is necessary for Impad1-mediated exocytosis, invasion, and metastasis • Inhibiting Impad1 or Syt11 represses lung cancer invasion and metastasis Metastasis remains the primary cause of cancer-related deaths, and a better understanding of the process will determine new therapeutic avenues. Bajaj et al. show that the Golgi apparatus and Golgi apparatus-associated proteins play a key role in determining tumor metastasis, indicating the importance of targeting the organelle during cancer. [ABSTRACT FROM AUTHOR]

BACKGROUND In this study, mould incidence and mycotoxin contamination were determined in freshly harvested maize samples collected from different agroclimatic regions of India. A total of 150 freshly harvested maize samples from major maize-growing areas of India (Karnataka, Andhra Pradesh and Tamilnadu) were collected during winter seasons 2010-2011 and 2011-2012 to determine their toxigenic fungal incidences, and mycotoxins were analyzed and quantified by high-perfomance liquid chromatography. A total of 288 fungal isolates comprising Fusarium, Aspergillus and Penicillium species were tested for aflatoxin B1 ( AFB1), ochratoxin A ( OTA), trichothecenes (deoxynivalenol ( DON) and T-2 toxin) and fumonisin B1 ( FB1). Chemotype determination of fungal isolates was carried out by molecular and chemical analysis through polymerase chain reaction ( PCR) and high-performance thin layer chromatography respectively. The diversity and distribution of the mycoflora among the studied samples were recorded in terms of frequency, density, importance value index and diversity indices. RESULTS A total of 288 fungal isolates were recovered from the 150 maize samples, of which 28 were positive for AFB1, 20 for OTA, 58 for FB1, 23 for DON and 11 for T-2 toxin chemotypes by PCR. Species-specific PCR assays were in line with morphological analysis. Toxigenic fungal incidences were found throughout the study region, and most of the toxins under study exceeded the maximum legal limits. The range of observed toxin concentrations were 48-58 µg AFB1, 76-123 µg FB1, 38-50 µg T-2, 72-94 µg DON and <5 µg OTA kg−1 grain sample. CONCLUSION Owing to the high incidences of toxigenic moulds and mycotoxins in the study area, there is a need for the creation of mycotoxin awareness among maize farmers of India to control the chronic adverse health effects on humans and livestock due to mycotoxins. © 2014 Society of Chemical Industry [ABSTRACT FROM AUTHOR]

Burkholderia pseudomallei, the causative agent of melioidosis has been recognized by CDC as a category B select agent. Although substantial efforts have been made for development of vaccine molecules against the pathogen, significant hurdles still remain. With no licensed vaccines available and high relapse rate of the disease, there is a pressing need for development of alternate protection strategies. Antibody-mediated passive protection is promising in this regard and our primary interest was to unravel this frontier of specific mAbs against Burkholderia pseudomallei infections, as functional characterization of antibodies is a pre-requisite to demonstrate them as protective molecules. To achieve this, we designed our study on in vitro-based approach and assessed two mAbs, namely BURK24 and BURK37, reactive with outer membrane proteins and lipopolysaccharide of the pathogen respectively, for their ability to manifest inhibitory effects on the pathogenesis mechanisms of B. pseudomallei including biofilm formation, invasion and induction of apoptosis. The experiments were performed using B. pseudomallei standard strain NCTC 10274 and a clinical isolate, B. pseudomallei 621 recovered from a septicemia patient with diabetic ailment. The growth kinetic studies of the pathogen in presence of various concentrations of each individual mAb revealed their anti-bacterial properties. Minimal inhibitory concentration and minimal bactericidal concentration of both the mAbs were determined by using standards of Clinical and Laboratory Standards Institute (CLSI) and experiments were performed using individual mAbs at their respective bacteriostatic concentration. As an outcome, both mAbs exhibited significant anti-Burkholderia pseudomallei properties. They limited the formation of biofilm by the bacterium and completely crippled its invasion into human alveolar adenocarcinoma epithelial cells. Also, the mAbs were appreciably successful in preventing the bacterium to induce apoptosis in A549 cells. The present study design revealed the protection attributes possessed by BURK24 and BURK37 that has to be further substantiated by additional in vivo studies. [ABSTRACT FROM AUTHOR]

BACKGROUND The aim of this study was to detect the presence of different mycotoxigenic Aspergillus species in major food grains from southern states of India, namely maize, paddy, groundnut and sorghum. A total of 200 isolates recovered from 320 grain samples from four southern states were tested for their toxin chemotypes using high-performance liquid chromatography ( HPLC), high-performance thin layer chromatography ( HPTLC) and polymerase chain reaction ( PCR) methods. The diversity and distribution of the isolates were recorded in terms of their frequency, density, importance value index and diversity indices. RESULTS Among the different grain samples tested, 83% of groundnut, 69% of maize, 57% of sorghum and 29% of paddy samples had aflatoxin B1 ( AFB1) levels above the allowed limit, while 82% of maize, 70% of sorghum, 42% of paddy and 17% of groundnut samples had ochratoxin A ( OTA) concentrations higher than the permitted threshold (5 µg kg−1). CONCLUSION Since the southern states of India are temperate regions, environmental factors, especially temperature and relative humidity, may be responsible for the high levels of mycotoxins present in the grains studied. Therefore there is a need to generate awareness among farmers and consumers about the possible adverse health effects of high levels of mycotoxins present in different food grains. © 2013 Society of Chemical Industry [ABSTRACT FROM AUTHOR]