Total extra-peritoneal approach to radical cystectomy with ureterostomy: A novel technique for the elderly and frail.

OBJECTIVES: Radical cystectomy with urinary diversion is the gold standard treatment for bladder cancer (high-risk/muscle invasive). The transperitoneal approach is associated with significant gastrointestinal complications like ileus. In the elderly and frail with a single functional kidney, we describe an extraperitoneal technique of radical cystectomy, with a ureterostomy, to be performed without general anesthesia. MATERIALS AND METHODS: The elderly, frail, and high-risk candidates for general anesthesia, with a prior history of nephroureterectomy with a second primary muscle-invasive bladder cancer, were chosen. All patients underwent the described procedure under combined spinal and epidural anesthesia. The posterior dissection was retrograde, caudal to cranial, with the peritoneum being opened only for resection of the dome. A cutaneous ureterostomy was fashioned on the side of the functional kidney. Peri-operative parameters were assessed for early recovery in this high-risk group. RESULTS: The mean age was 82 years (range: 73-91), with Charleson Comorbidity Index 5, and were all deemed unfit for neoadjuvant chemotherapy. With a median duration of 127.5 minutes, an average blood loss of 225ml, and no patient requiring general anesthesia; early ambulation, early return of bowel function, and a lesser hospital stay (7 days) with minimal morbidity were achieved. Negative surgical margins were achieved in all cases, with a mean harvest of 29 lymph nodes. Only 1 patient developed stomal stenosis. The cause-specific survival (CSS) is 100% at 2 years. CONCLUSIONS: The highlighting features are the early return of bowel function (flatus passage on day 1) and the avoidance of the cardio-pulmonary complications of general anesthesia. The extraperitoneal cystectomy offers a promising alternative in this select group and warrants further studies to extrapolate this technique for bilateral urinary drainage.

Sarcomatoid Carcinoma Metastasis to the Colon from a Small Renal Mass: Case Report with Review of Literature.

A third of patients with renal cell carcinoma (RCC) present with metastatic disease. Metastasis in RCC from small renal mass (SRM) (≤4 cm) is rare. We report a case of stage cT1a clear-cell RCC with low-risk features on pathology presenting with disproportionately large synchronous solitary metastasis to the transverse colon. He underwent resection of the mass with the involved transverse colon and adjoining mesocolon. Intestinal continuity was restored, following which partial nephrectomy was performed for the right renal tumor. Final pathology of the right renal mass confirmed clear-cell RCC. The large mass after immunohistochemistry profile confirmed metastasis from the renal tumor.

Conditional survival in patients treated for gastric cancer with a curative intent.

BACKGROUND AND OBJECTIVES: Conditional survival (CS) provides the probability that a patient who has already survived a certain number of years after treatment will survive an additional number of years. We aim to study the CS of patients with gastric cancer. METHODS: Patients who underwent curative intent treatment for gastric cancer in a single institution between 2007 and 2018 were included in the analysis. The probability (CS) that a patient who has already survived x years will survive an additional y year, was calculated as CS (y/x) = S(x + y)/S(x). RESULTS: The probability of surviving an additional 3 years if a patient had already survived 1, 2, 3, 4, and 5 years after treatment were 64.2%, 74.5%, 81.6%, 83.2%, and 88.2%, respectively whereas the 4-, 5-, 6-, 7-, and 8-year actuarial OS were only 47.2%, 43.2%, 41%, 39.4%, and 38.2%, respectively. The independent prognostic factors associated with poor survival were age >60 years, T stage ≥T3, N stage ≥N2, proximal tumor location, and lymph node ratio > 0.18. Patients with these high-risk features showed the greatest increase in CS3 over time. CONCLUSION: CS estimates provided a more dynamic prognostic information over time for patients treated for gastric cancer with curative intent.

Cyanobacteria-specific algicidal mechanism of bioinspired naphthoquinone derivative, NQ 2-0.

To mitigate cyanobacterial blooms, the naphthoquinone derivative, NQ 2-0, which has selective algicidal activity against cyanobacteria, has been developed. However, due to a lack of information on its algicidal mechanisms, there are significant gaps in our understanding of how this substance is capable of selectively killing cyanobacteria. Here, we investigated the selective algicidal mechanisms of NQ 2-0 using target (Microcystis aeruginosa) and non-target (Cyclotella sp. and Selenastrum capricornutum) species. NQ 2-0 showed selective algicidal activity against only M. aeruginosa, and this activity was strongly light-dependent. This NQ compound has selectively reduced the oxygen evolution rate and photosystem II (PSII) efficiency of M. aeruginosa throughout blocking electron transfer from the photosynthetic electron transport system, and significantly (p ≤ 0.05) increased levels of reactive oxygen species (ROS), resulting in membrane damage through lipid peroxidation. In ultrastructural observations, thylakoid membranes were disintegrated within 12 h after NQ 2-0 treatment, and cytoplasmic vacuolation and disintegrated cellular membrane were observed at 24 h. These findings suggest that increased ROS levels following NQ 2-0 treatment may induce cell death. Interestingly, compared to non-target eukaryotic cells, M. aeruginosa showed relatively late antioxidant response to reduce the increased ROS level, this may enhance algicidal activity against this cyanobacterium.

Correction to: Pelagibaca bermudensis promotes biofuel competence of Tetraselmis striata in a broad range of abiotic stressors: dynamics of quorum-sensing precursors and strategic improvement in lipid productivity.

[This corrects the article DOI: 10.1186/s13068-018-1097-9.].

Pelagibaca bermudensis promotes biofuel competence of Tetraselmis striata in a broad range of abiotic stressors: dynamics of quorum-sensing precursors and strategic improvement in lipid productivity.

BACKGROUND: Amelioration of biofuel feedstock of microalgae using sustainable means through synthetic ecology is a promising strategy. The co-cultivation model (Tetraselmis striata and Pelagibaca bermudensis) was evaluated for the robust biofuel production under varying stressors as well as with the selected two-stage cultivation modes. In addition, the role of metabolic exudates including the quorum-sensing precursors was assessed. RESULTS: The co-cultivation model innovated in this study supported the biomass production of T. striata in a saline/marine medium at a broad range of pH, salinity, and temperature/light conditions, as well as nutrient limitation with a growth promotion of 1.2-3.6-fold. Hence, this developed model could contribute to abiotic stress mitigation of T. striata. The quorum-sensing precursor dynamics of the growth promoting bacteria P. bermudensis exhibited unique pattern under varying stressors as revealed through targeted metabolomics (using liquid chromatography-mass spectrometry, LC-MS). P. bermudensis and its metabolic exudates mutually promoted the growth of T. striata, which elevated the lipid productivity. Interestingly, hydroxy alkyl quinolones independently showed growth inhibition of T. striata on elevated concentration. Among two-stage cultivation modes (low pH, elevated salinity, and nitrate limitation), specifically, nitrate limitation induced a 1.5 times higher lipid content (30-31%) than control in both axenic and co-cultivated conditions. CONCLUSION: Pelagibaca bermudensis is established as a potential growth promoting native phycospheric bacteria for robust biomass generation of T. striata in varying environment, and two-stage cultivation using nitrate limitation strategically maximized the biofuel precursors for both axenic and co-cultivation conditions (T and T-PB, respectively). Optimum metabolic exudate of P. bermudensis which act as a growth substrate to T. striata surpasses the antagonistic effect of excessive hydroxy alkyl quinolones [HHQ, 4-hydroxy-2-alkylquinolines and PQS (pseudomonas quorum signal), 2-heptyl-3-hydroxy-4(1H)-quinolone].

Revealing the distinct habitat ranges and hybrid zone of genetic sub-populations within Pseudo-nitzschia pungens (Bacillariophyceae) in the West Pacific area.

Genetic sub-populations (clades) of cosmopolitan marine diatom Pseudo-nitzschia pungens might have distinct habitats, and their hybrid zone is suspected in higher latitude area of the West Pacific area, however, it is still unrevealed because of technical difficulties and lack of evidences in natural environments. The aim of this study is to investigate the habitat characteristics of each clade of P. pungens on geographical distribution with the habitat temperature ranges of each clade and to reveal their hybrid zone in the West Pacific area. We employed the 137 number of nucleotide sequences of P. pungens and its sampling data (spatial and temporal scale) originated from the West Pacific area, and used field application of qPCR assay for intra-specific level of P. pungens. Only two genotypes, clade I and III, were identified in the West Pacific area. Clade I was distributed from 39 to 32.3°N, and clade III were from 1.4 to 34.4°N. The estimated habitat temperature for the clade I and clade III ranges were 8.1-26.9 °C and 24.2-31.2 °C, respectively. The latitudinal distributions and temperature ranges of each clade were significantly different. The qPCR assay employed, and results suggested that the hybrid zone for clade I and III has been observed in the southern Korean coasts, and clade III might be introduced from the Southern Pacific area. The cell abundances of clade III were strongly related with the higher seawater temperature and warm current force. This study has defined distinct habitat characteristics of genetically different sub-populations of P. pungens, and revealed its hybrid zone in natural environment for the first time. We also provided strong evidences about dispersion of the population of clade III to higher latitude in the West Pacific area.

Ecological assessment of an algaecidal naphthoquinone derivate for the mitigation of Stephanodiscus within a mesocosm.

The novel eco-friendly algaecidal naphthoquinone derivate was used to control harmful algal bloom causing species Stephanodiscus and, its effect was assessed on other undesired and non-targeted microbial communities. We conducted a mesocosm experiment to investigate the effects of this novel algaecide on native microbial communities rearing in water collected from Nakdonggang River. Upon treatment of the mesocosm with the naphthoquinone derivate the concentration of Chl-a decreased from 20.4 µg L-1 to 9.5 µg L-1 after 2 days. The turbidity has also shown decrement (exhibited 15.5 NTU on the 7th day). The concentrations of DOC and phosphate in the treatment were slightly higher than those in the control due to the decomposition of dead Stephanodiscus, whereas the DO and pH in the treated condition were slightly lower than those in the control; which was due to increment of organic acids and higher degradation activity. Results showed that bacterial abundance were not significantly different but community composition were slightly different as revealed by NGS (Next generation sequencing). The variation in HNF (Heterotrophic nanoflagellates) revealed that the bacterial community composition changed following the change in bacterial abundance. During the treatment, the abundance of Stephanodiscus was significantly reduced by more than 80% after 6 days, and the abundance of ciliates and the dominant species, Halteria grandinella, had shown marked decline. The abundance of zooplankton sharply decreased to 5 ind. L-1on the 8th day but increased again by the end of the study period. The Shannon-Wiener diversity index of phytoplankton, ciliates and zooplankton in the treated mesocosm increased significantly after 4, 7 and 8 days, respectively. The marked changes in the ecosystem structure were observed in treatment compare to control. However, the beneficial microalgal populations were not affected which indicated possibility of restoration of treated ecosystem and regain of healthy community structure after certain period.

Development of a qPCR assay for tracking the ecological niches of genetic sub-populations within Pseudo-nitzschia pungens (Bacillariophyceae).

Three genetic sub-populations (clade I, II and III) of Pseudo-nitzschia pungens, the potential toxic marine diatom, are known to have distinguishable growth characteristics under different culture conditions and distinct distributed patterns in the world. However, to date their exact eco-physiological traits are unrevealed in fields due to lack of the method to detect and/or measure abundances of each sub-populations, hence, the qPCR (quantitative real-time polymerase chain reaction) assay was developed to detect and quantify the P. pungens cells of each clade. Designed two specific primer sets, Pcla12F/R (for clade I and II) and Pcla3F/R (for clade III) only could amplify each target genomic DNA. The, significant linear relationships (R2>0.998) was established between Ct (threshold cycle) value and the log of cell abundance for each clade. Through the melting curve analysis, comparisons for gene copy numbers among the three clades and spike test for field study, our qPCR assay was reliable to quantify the cell numbers of each clade. There was strong linear correlation (R2>0.990) between cell abundances as estimated by qPCR assay and direct counting via light microscope in spike test, and 0.24 (clade I), 0.25 (clade II) and 0.33 (clade III) P. pungens cells per mL were detected markedly upon the use of specific two-primer set. Finally, developed qPCR assay was applied on field samples successfully. Our study implicate that our qPCR assay is an accurate and sensitive technique to estimate the cell abundances of each clade of P. pungens in field works.

Phycospheric Native Bacteria Pelagibaca bermudensis and Stappia sp. Ameliorate Biomass Productivity of Tetraselmis striata (KCTC1432BP) in Co-cultivation System through Mutualistic Interaction.

Effective sustainable algal cultivation techniques are essential for mass production of the marine microalga Tetraselmis for biofuel and array of co-products. The phycospheric communities affect the microalgal growth and metabolism through various allelochemical and nutrient interactions; hence, their potential to affect the quantity and quality of both biomass and bioproducts is significant. In the present study, we have screened the phycospheric communities of biofuel producing Tetraselmis striata (KCTC1432BP). A total of 26 bacterial strains were isolated and identified from the phycosphere of T. striata mass culture. Then, each bacterial strain was tested in co-cultivation conditions with T. striata for evaluating its growth promoting and inhibitory effects. Among these all strains, two promising strains (Pelagibaca bermudensis KCTC 13073BP and Stappia sp. KCTC 13072BP) were selected because of their maximum growth promoting effects and mutualistic interactions. The growth rate, biomass productivity, lipid contents, and fatty acids were analyzed during their combined growth in O3 media and compared with axenic growth of T. striata. Later, growth promoting mechanisms in the co-cultivation environment were investigated for these promising bacterial strains under replete and limited conditions of nutrients (nitrate, phosphate, and vitamin B12). The growth promoting potential of P. bermudensis was illustrated by the two fold enhancement in biomass productivity. These bacteria are promising for microalgal cultivation without any negative effects on the native seawater bacterial communities, as revealed by next generation sequencing analysis. This study represents, to date, the first report highlighting the role of phycospheric growth promoting bacteria of promising biofuel feedstock T. striata.

Cultivation of Nannochloropsis oceanica biomass rich in eicosapentaenoic acid utilizing wastewater as nutrient resource.

The eicosapentaenoic acid rich marine eustigmatophyte Nannochloropsis oceanica was grown in wastewaters sampled from four different industries (i.e. pesticides industry, pharmaceutical industry, activated sludge treatment plant of municipality sewage and petroleum (oil) industry). Under the wastewater based growth conditions used in this study, the biomass productivity ranged from 21.78±0.87 to 27.78±0.22mgL(-1)d(-1) in relation to freeze dried biomass, while the lipid productivity varied between 5.59±0.02 and 6.81±0.04mgL(-1)d(-1). Although comparatively higher biomass, lipid and EPA productivity was observed in Conway medium, the %EPA content was similarly observed in pesticides industry and municipal effluents. The results highlight the possibility of selectively using wastewater as a growth medium, demonstrating the elevated eicosapentaenoic acid content and biodiesel properties, that complies with the European standards for biodiesel.

Integrated process of two stage cultivation of Nannochloropsis sp. for nutraceutically valuable eicosapentaenoic acid along with biodiesel.

The marine eustigmatophyte Nannochloropsis is one of the potential producers of eicosapentaenoic acid (EPA), a valued nutraceutical. Nannochloropsis sp. was cultivated under photoautotrophic condition utilizing CO2 in a two phase cultivation process in order to enhance the eicosapentaenoic acid (EPA) productivity. It was cultivated in a photobioreactor up to late log phase for cell growth (phase I). Then, the culture was harvested and confronted to relatively low temperature (10 °C) and low light (30 µmol photons m(-2) s(-1)) in both photobioreactor and Erlenmeyer flask (phase II), thus augmenting EPA% by 3.4 fold. Lower temperature with low light favored the synthesis of EPA although, biomass productivity, lipid content and lipid productivity were slightly decreased relative to phase I. The total lipids extracted from Nannochloropsis sp. fractionated into neutral lipids (NLs), glycolipids (GLs) and phospholipids (PLs) and a major proportion of EPA was found in phospholipids. Results suggested that low temperature and low light may ameliorate partitioning towards EPA in phospholipids.

Salinity induced oxidative stress enhanced biofuel production potential of microalgae Scenedesmus sp. CCNM 1077.

Microalgal biomass is considered as potential feedstock for biofuel production. Enhancement of biomass, lipid and carbohydrate contents in microalgae is important for the commercialization of microalgal biofuels. In the present study, salinity stress induced physiological and biochemical changes in microalgae Scenedesmus sp. CCNM 1077 were studied. During single stage cultivation, 33.13% lipid and 35.91% carbohydrate content was found in 400 mM NaCl grown culture. During two stage cultivation, salinity stress of 400 mM for 3 days resulted in 24.77% lipid (containing 74.87% neutral lipid) along with higher biomass compared to single stage, making it an efficient strategy to enhance biofuel production potential of Scenedesmus sp. CCNM 1077. Apart from biochemical content, stress biomarkers like hydrogen peroxide, lipid peroxidation, ascorbate peroxidase, proline and mineral contents were also studied to understand the role of reactive oxygen species (ROS) mediated lipid accumulation in microalgae Scenedesmus sp. CCNM 1077.

Dominance of cyanobacterial and cryptophytic assemblage correlated to CDOM at heavy metal contamination sites of Gujarat, India.

Industrial clusters of Gujarat, India, generate high quantity of effluents which are received by aquatic bodies such as estuary and coastal water. In the present study, microalgal assemblage, heavy metals, and physico-chemical variables were studied from different habitats. Principal component analysis revealed that biovolume of cyanobacterial and cryptophytic community positively correlated with the heavy metal concentration (Hg, As, Zn, Fe, Mo, Ni, and Co) and chromophoric dissolved organic matter (CDOM) under hypoxic environment. Green algae and diatoms dominated at comparatively lower nitrate concentration which was positively associated with Pb and Mn.

Potential of Monoraphidium minutum for carbon sequestration and lipid production in response to varying growth mode.

Mixotrophic growth at flask level and, autotrophic-mixotrophic and autotrophic growth in photobioreactor by utilizing CO2/air/flue gas were checked for the isolated strain of Monoraphidium minutum from polluted habitat. Our study confirmed that it is a saturated fatty acid rich (30.92-68.94%) microalga with lower degree of unsaturation oil quality (42.06-103.99) making it potential biodiesel producing candidate. It showed encouraging biomass productivity (80.3-303.8mgl(-1)day(-1)) with higher total lipid (22.80-46.54%) under optimum glucose, fructose, microalgal biodiesel waste residue and sodium acetate fed mixotrophic conditions. The pH control by intermittent CO2, continuous illumination with 30% flue gas, and utilization of biodiesel glycerin were effective schemes to ameliorate either biomass productivity or % lipids or both of these parameters at photobioreactor scale (7.5L working volume). The modulation of environmental variables (pH control, CO2 and organic substrates concentration) could augment % saturated fatty acids, such as C16:0.

Fatty acids as biomarkers of microalgae.

Microalgae are primary producers of the food chain and hold prominence towards pharmaceutical and nutraceutical applications. Fatty acids (FAs) are one of the primary metabolites of microalgae, which enrich their utility both in the form of food and fuels. Additionally, the vast structural diversity coupled with taxonomic specificity makes these FAs as potential biomarkers. The determination of lipid and fatty acid profiling of 12 different strains of microalgae has been accomplished in this study and further discussed in respect to their chemotaxonomic perspective in microalgae. Palmitic acid (C16:0) and oleic acid (C18:1n9c) were found to be dominant among the members of Cyanophyceae whereas members of Chlorophyceae were rich in palmitic acid (C16:0), oleic acid (C18:1n9c) and linoleic acid (C18:2n6). The application of principal component analysis (PCA) and algorithmic hierarchical clustering (AHC) resulted in the segregation of the studied microalgal strains into 8 different orders belonging to 2 distinct phyla according to their phylogenetic classification. Nutritionally important FAs like eicosapentaenoic acid (EPA, C20:5n3) and docosahexaenoic acid (DHA, C22:6n3) were detected only in Chlorella sp. belonging to Chlorophyceaen family. Differential segregation of microalgae with respect to their fatty acid profile indicated the potential utility of FAs as biomarkers.

Effect of light quality on the C-phycoerythrin production in marine cyanobacteria Pseudanabaena sp. isolated from Gujarat coast, India.

The isolated cyanobacterium containing biopigments like chlorophyll-a, phycoerythrin, phycocyanin, and carotenoid was cultured under different quality of light modes to ascertain biomass and pigment productivity. On the basis of 16S rRNA gene sequence, the isolate was identified as Pseudanabaena sp. Maximum biomass concentration obtained in white-, blue-, and green-light was 0.82, 0.94, and 0.89 g/L, respectively. It was observed that maximum phycoerythrin production was in green light (39.2 mg/L), ensued by blue light (32.2 mg/L), while phycocyanin production was maximum in red light (10.9 mg/L). In yellow light, pigment production as well as the growth rate gradually declined after 12 days. Carotenoid production decreased in blue-, white-, and red-light after 15 days, while in green light it had increased gradually. The present communication suggests that Pseudanabaena sp. can be used for commercial production of phycoerythrin when grown under green light.