Production Levels Research Articles

Fast and robust recombinant protein production utilizing episomal stable pools in WAVE bioreactors

Protein reagents are essential resources for several stages of drug discovery projects from structural biology and assay development through lead optimization. Depending on the aim of the project different amounts of pure protein are required. Small-scale expressions are initially used to determine the reachable levels of production and quality before scaling up protein reagent supply. Commonly, amounts of several hundreds of milligrams to grams are needed for different experiments, including structural investigations and activity evaluations, which require rather large cultivation volumes. This implies that cultivation of large volumes of either transiently transfected cells or stable pools/stable cell lines is needed. Hence, a production process that is scalable, speeds up the development projects, and increases the robustness of protein reagent quality throughout scales. Here we present a protein production pipeline with high scalability. We show that our protocols for protein production in Chinese hamster ovary cells allow for a seamless and efficient scale-up with robust product quality and high performance. The flexible scale of the production process, as shown here, allows for shorter lead times in drug discovery projects where there is a reagent demand for a specific protein or a set of target proteins.

Abstract B006: GSTT2 modulates immune activation and impacts response to BCG immunotherapy in bladder cancer

Abstract Whilst M. bovis Bacillus Calmette-Guérin (BCG) therapy is the gold-standard for treating high risk non-muscle invasive bladder cancer (NMIBC), 30-40% of patients fail therapy, resulting in disease recurrence and progression. Loss of glutathione-S-transferase theta 2 (GSTT2) expression resulting from a promoter deletion, has been associated with modulation of intracellular ROS and BCG survival in BC cells. Retrospective analysis of NMIBC patients revealed that patients with the promoter deletion responded better to fewer instillations of BCG. To understand these responses, wildtype (WT) and GSTT2-knockout (KO) mice were implanted with MB49-PSA BC cells, either subcutaneously or orthotopically, and treated with weekly BCG instillations. Subcutaneous tumors were harvested after 3 BCG instillations for flow cytometry analysis, or after 4 BCG instillations for immunofluorescence (IF) imaging and gene expression analysis. Bladders (orthotopic model) were harvested after 4 BCG instillations for single-cell RNA sequencing. Splenocytes from WT and KO male mice were also stimulated in-vitro with TLR agonists, for 48h, to assess immune responses. In the subcutaneous model, in male mice, the cure rate was higher in WT mice (57%) compared to KO mice (0%). Flow cytometry analysis showed significantly higher numbers of tumor-associated CD8+ T-cells in BCG-treated KO males compared to WT males; however, IF imaging revealed higher penetration of CD8+ T-cells in tumors from WT males. In female mice, the cure rate was similar in WT (25%) and GSTT2-KO (38%) mice. IF imaging showed higher penetration of CD8+ T-cells in KO females. Quantitative real-time PCR (q-PCR) analysis of BCG-treated subcutaneous tumors from female mice revealed lower expression of the immune exhaustion markers, PD-L1 and CTLA-4, in KO mice compared to WT mice. Similarly, in the orthotopic model (female mice), q-PCR analysis of bladders also showed lower expression of PD-L1 and CTLA-4 in KO mice compared to WT mice. Single-cell RNA sequencing of bladders from BCG-treated mice revealed differences in the expression of genes such as Peak1, Hmga2, Scl2a1, Kras, Ahank, Gsta4, Mecom and Sh3gl2, of which Gsta4, Mecom, Sh3gl2, Hmga2 and Peak1 were validated using q-PCR. In-vitro analysis of splenocytes from male mice revealed higher levels of IL-6, IL-1β and IFN-α, after stimulation with TLR7/8 and TLR9 agonists, in KO splenocytes relative to WT splenocytes. Despite the higher levels of cytokine production by splenocytes and higher proportion of tumor-associated CD8+ T-cells in KO male mice, these mice fared poorly, which could be explained by immune exhaustion or lack of immune penetration into the tumor. In females, lower expression of immune exhaustion markers in KO mice did not yield better responses; however, single-cell data showed differences in expression of genes associated with pro- and anti-tumorigenic pathways, which could influence response. Thus, loss of GSTT2 may influence the response to BCG therapy, through modulating downstream signaling pathways and immune responses. Citation Format: Mugdha Vijay Patwardhan, Kane Toh, Edmund Chiong, Ratha Mahendran. GSTT2 modulates immune activation and impacts response to BCG immunotherapy in bladder cancer [abstract]. In: Proceedings of the AACR Special Conference on Bladder Cancer: Transforming the Field; 2024 May 17-20; Charlotte, NC. Philadelphia (PA): AACR; Clin Cancer Res 2024;30(10_Suppl):Abstract nr B006.

Effects of single or combined feeding with <i>Bacillus subtilis</i> and organic acid mixture on egg production, physiological responses, and cholesterol level in chicken eggs

Effects of single or combined feeding with <i>Bacillus subtilis</i> and organic acid mixture on egg production, physiological responses, and cholesterol level in chicken eggs

A colony health and economic comparison of Varroa-resistant Varroa destructor (Mesostigmata: Varroidae) and commercial honey bees (Hymenoptera: Apidae).

Honey bees (Apis mellifera L.) are the premier agricultural pollinators with direct ecological value and are key to some agro-economies. Major factors have negatively impacted honey bee health in the past 2 decades with Varroa (Varroa destructor Anderson and Trueman) infestation rising as a principal predictor of colony mortality. A key strategy deployed in Varroa management is breeding for resistant honey bee populations that can maintain comparable levels of productivity as nonresistant populations. In this study, we examine one such population, Hilo honey bees, within the context of a common garden contrast with a commercial population in a stationary honey production operation. We compare colony survival, health, yield, and profit outcomes to show how this specific breeding population retains a profit value in honey production operations while maintaining higher survival and lower Varroa infestation levels than the commercial population. This information can be used by commercial beekeepers to make best management practice decisions and inspire further work examining what trade-offs, if any, are present in this Varroa-resistant population.

Novel insights into presenilin 1 mutation associated with a distinctive dementia phenotype and cotton wool plaques.

The mutations in the presenilin 1 gene (PSEN1) are the main cause of familial Alzheimer's disease. PSEN1 mutations affect amyloid-beta peptide production, which accumulates in the brain as senile plaque and cotton wool plaques (CWPs) and relates to other neurodegenerative disorders. Here we report the second case of the PSEN1 G266S mutation, which showed distinctive neuropathological features, including abundant CWPs. Lewy body pathology, and altered amyloid-beta production. Using the proband's samples, we performed genetic analysis of the PSEN1, APP, MAPT, and APOE genes, histopathological and immunohistochemical analysis of the brain tissue, and biochemical analysis of Aβ production in COS cells transfected with wild-type or mutant PSEN1. The patient presented with memory loss, abnormal behavior, and visual hallucinations. Brain scans showed reduced blood flow, mild atrophy, and white matter lesions. Genetic analysis revealed a heterozygous mutation at codon 266 (G266S) of PSEN1 and polymorphism of MAPT (Q230R). The brain had many CWPs, severe cerebral amyloid angiopathy (CAA), senile plaque, Lewy bodies, and neurites. Electron microscopy displayed myelinated fiber degeneration, mitochondrial damage, and amyloid fibrils in the white matter. The production level of Aβ42 in PSEN1 G266S-transfected cells significantly increased. Our findings suggest that the PSEN1 G266S mutation may cause a heterogeneous clinical and pathological phenotype, influenced by other genetic or environmental factors.

Do No Harm? Unintended Consequences of Pharmaceutical Price Regulation in India

The Drugs (Prices Control) Order of 2013 in India regulated the prices of certain essential and life-saving drugs to ensure their affordability and availability, expecting an increase in sales of those drugs. The authors study the effects of the regulation on sales volumes of each regulated drug using a synthetic control approach with sales data from a comparable country that did not experience a regulatory change. They assess the robustness of the results via multiple empirical approaches to triangulate the findings. Contrary to the order's objectives, they find that sales volumes declined for regulated drugs. Since the order placed restrictions on production levels and on drugs exiting the market, the lowered margins of regulated drugs could have pushed pharmaceutical firms to reduce their marketing expenditures on them. The authors provide evidence of such a reduction using detailing data from a large pharmaceutical firm. The findings illustrate that this shift in detailing adversely affected prescriptions from physicians without formal medical degrees who treat poor and disadvantaged people in India—patients that the order was intended to help the most. A survey the authors conducted shows that these physicians rely on detailing more than medically trained doctors. Taken together, the results provide insights into the strategic actions of firms when faced with regulations, and highlight their unintended consequences. The generalizable nature of the study's findings across a broad set of medications has implications for governmental agencies in terms of the need to account for the entire ecosystem when implementing such regulations.

Selenium nanoparticles decorated with polysaccharides from Sargassum fusiforme protects against 6-OHDA-induced neurotoxicity in PC12 cells and rat model of Parkinson's disease

Parkinson's disease (PD) is a neurodegenerative disorder and identifying disease-causing pathways and drugs that target them has remained challenging. Herein, selenium nanoparticles decorated with polysaccharides from Sargassum fusiforme (SFPS-SeNPs) were investigated on 6-OHDA-induced neurotoxicity in PC12 cells and rats. 6-OHDA can significantly increase neurotoxicity, oxidative stress and decrease the activity of superoxide dismutase (SOD) and glutathione peroxidase (GPx) both in vitro and vivo. In vitro, treatment with SFPS-SeNPs can significantly decrease 6-OHDA cytotoxicity, reactive oxygen species (ROS) production or malondialdehyde (MDA) levels, and cell apoptosis, significantly increased the activity of SOD and GPx. In vivo, 6-OHDA exposure could also decrease the expression of Nrf2 and OH-1, while treatment with SFPS-SeNPs (1 mg Se/kg) increased. SFPS-SeNPs can protect neurons from 6-OHDA-induced neurotoxicity by regulating apoptosis and Nrf2/ARE pathway. The present study demonstrated that SFPS-SeNPs is a good candidate for developing a new drug against neurodegenerative diseases such as PD.

Consumer Awareness and Perceptions of Pesticide Residues in Vegetables in Bangladesh

Vegetables are heavily sprayed and prone to retaining residues. The Department of Agriculture Extension (DAE), Bangladesh has set up some projects to disseminate technologies among the farmers to reduce pesticide residues. Farmers are motivated to produce pesticide-free vegetables using vermin compost and other organic fertilizers. The study investigates consumers’ awareness and perceptions of pesticide residues in vegetables. A survey was conducted at the local market in Mymensingh, Gazipur, and Dhaka city. In total. 200 sample respondents were collected. Data were analyzed using descriptive methods. There was a substantial heterogeneity in preferences across consumers. The consumers were aware of the health risks associated with vegetables produced using pesticides and chemical fertilizers. More than 60 percent of respondents thought that vegetables are usually contaminated at the farm or production level using excessive pesticides. Most of the consumers knew the effects of pesticide-borne health risks and suggested to ban some pesticides and monitoring the use of others. Consumers considered pesticide residue-free vegetables as being safer, healthy, no harmful effects, better taste, more nutritive value, and good for the environment, and would like to pay a premium for those products. Media particularly television and newspapers can play an important role in creating awareness about food safety among the producers and consumers. Government programmes on food safety should be channeled through television and newspapers where the majority of the people get their information. Policy measures should include creating awareness concerning the relevance of production, and consumption through effective marketing strategies and educational awareness. Res. Agric. Livest. Fish. Vol. 11, No. 1, April 2024: 25-33

Economic Efficiency of Agriculture in Punjab: Evidence from Cost of Cultivation Survey

Understanding the economic efficiency of agriculture production in Punjab is an interest of this study. The Cost of Cultivation Survey data set for the block period 2014–15 to 2016–17 has been used. Data envelopment analysis deterministic approach has been applied to measure production efficiency. The estimation showed that in the case of wheat, 73 per cent of tehsils and in the case of paddy, 77 per cent of tehsils in Punjab are found to be inefficient in crop production due to poor input utilisation (managerial inefficiency) and scale size (scale inefficiency). But the higher proportion of inefficiency is mainly attributed to the non-optimal scale of production as compared to pure technical allocation. The farmers can increase their production level on average by 7–17 per cent through improving farm efficiency. Findings showed that farmers are allocating their input resources in an exploitative manner due to which they failed to use an appropriate combination of inputs, which is necessary to achieve cost minimisation. These findings have important implications for agricultural development policy and are likely to offer some useful insights for the adequate use of scarce sources. JEL Codes: C33, C67, Q10, Q15

Accounting for product recovery potential in building life cycle assessments: a disassembly network-based approach

PurposeExisting life cycle assessment (LCA) methods for buildings often overlook the benefits of product recovery potential, whether for future reuse or repurposing. This oversight arises from the limited scope of such methods, which often ignore the complex interdependencies between building products. The present paper, backed by its supplementary Python library, introduces a method that addresses this gap, emphasizing the influence of product interdependencies and future recovery potential on environmental impact.MethodsImplementing the proposed method requires adding a phase, the recovery potential assessment, to the four phases that constitute an LCA according to the ISO 14040/14044 guidelines. Given the disassembly sequence for each product, in the first step of the recovery potential assessment, a disassembly network (DN) is created that displays structural and accessibility dependencies. By calculating the average of the disassembly potential (DP) of each structural dependency (second step) associated with that product, we obtain the DP (0.1–1) at the product level in a third step. Because there is no empirical data available to support a specific relationship between product disassembly potential and recovery potential (RP) (0–1), we employ, in a fourth step, a flexible model specification to represent scenarios of how this relationship may look like. Ultimately, for each scenario, the resulting RP is used to enable a probabilistic material flow analysis with a binary outcome, whether to be recovered or not. The resulting product-level median material flows are then used to quantify the building’s environmental impact for a given impact category in the life cycle impact assessment (LCIA). The results are interpreted through an uncertainty, hotspot, and sensitivity analysis.Results and discussionOur results show that not considering the interdependencies between building products in building LCAs results in underestimating the embodied greenhouse gas (GHG) emissions by up to 28.29%. This discrepancy is primarily attributed to a failure to account for additional material flows stemming from secondary replacements owing to the interdependencies during the life cycle. When accounting for end-of-life recovery benefits, a zero-energy building (ZEB) design incorporating some DfD principles demonstrated up to 45.94% lower embodied GHG emissions than the ZEB design with low disassembly potential when assuming that recovered products will be reused.ConclusionsOur approach provides first-of-a-kind evidence that not accounting for recovery potential may significantly distort the results of an LCA for buildings. The method and its supporting code support the semi-automated calculation of the otherwise neglected potential environmental impact, thus helping to drive the transition towards a more sustainable built environment. The supporting code allows researchers to build on the proposed framework if more data on the relationship between DP and RP become available in the future. Finally, while applied to buildings in this paper, the proposed framework is adaptable to any complex product with limited modifications in the supporting code.

Thrivers and Survivors during Study Abroad: The Individual Cases of Japanese Learners of English

Whilst study abroad (SA) periods hold much promise for foreign language development, increasing research suggests sojourners’ experiences are unique, and language development does not always follow a linear trajectory. For some learners, SA has little impact on their language performance despite the affordances of L2 immersion (the Survivors). Other learners maximise the potential of SA, and this has a positive impact on their language development (the Thrivers). This paper examines the selected cases of four Japanese learners of English and their request language performance during a 10-month SA in the UK. Changes in pragmatic knowledge (based on appropriateness ratings) were documented at three equidistant time points. Language contact profile data also provided quantitative insights into the learners’ extracurricular language use and qualitative personal reflections. The selected cases illustrate two learners surviving the SA experience, showing minimal change in their request performance. The other two learners thrived during SA, showing accelerated performance in terms of lexical variation at the production level. This paper reports on the case histories of these learners to better understand these unique experiences and pragmatic discrepancies. Suggestions for how learners might be more pragmatically successful during SA are also offered.

Citrate-buffered Yamanaka medium allows to produce high-yield bacterial nanocellulose in static culture using Komagataeibacter strains isolated from apple cider vinegar.

Bacterial nanocellulose (BNC) is a sustainable, renewable, and eco-friendly nanomaterial, which has gained great attentions in both academic and industrial fields. Two bacterial nanocellulose-producing strains (CVV and CVN) were isolated from apple vinegar sources, presenting high 16S rRNA gene sequence similarities (96%-98%) with Komagataeibacter species. The biofilm was characterized by scanning electron microscopy (SEM), revealing the presence of rod-shaped bacteria intricately embedded in the polymeric matrix composed of nanofibers of bacterial nanocellulose. FTIR spectrum and XRD pattern additionally confirmed the characteristic chemical structure associated with this material. The yields and productivities achieved during 10 days of fermentation were compared with Komagataeibacter xylinus ATCC 53524, resulting in low levels of BNC production. However, a remarkable increase in the BNC yield was achieved for CVV (690% increase) and CVN (750% increase) strains at day 6 of the fermentation upon adding 22mM citrate buffer into the medium. This effect is mainly attributed to the buffering capacity of the modified Yakamana medium, which allowed to maintain pH close to 4.0 until day 6, though in combination with additional factors including stimulation of the gluconeogenesis pathway and citrate assimilation as a carbon source. In addition, the productivities determined for both isolated strains (0.850 and 0.917gL-1 d-1) compare favorably to previous works, supporting current efforts to improve fermentation performance in static cultures and the feasibility of scaling-up BNC production in these systems.

Microwave-Assisted Synthesis of SnO2@ZnIn2S4 Composites for Highly Efficient Photocatalytic Hydrogen Evolution.

This research successfully synthesized SnO2@ZnIn2S4 composites for photocatalytic tap water splitting using a rapid two-step microwave-assisted synthesis method. This study investigated the impact of incorporating a fixed quantity of SnO2 nanoparticles and combining them with various materials to form composites, aiming to enhance photocatalytic hydrogen production. Additionally, different weights of SnO2 nanoparticles were added to the ZnIn2S4 reaction precursor to prepare SnO2@ZnIn2S4 composites for photocatalytic hydrogen production. Notably, the photocatalytic efficiency of SnO2@ZnIn2S4 composites is substantially higher than that of pure SnO2 nanoparticles and ZnIn2S4 nanosheets: 17.9-fold and 6.3-fold, respectively. The enhancement is credited to the successful use of visible light and the facilitation of electron transfer across the heterojunction, leading to the efficient dissociation of electron-hole pairs. Additionally, evaluations of recyclability demonstrated the remarkable longevity of SnO2@ZnIn2S4 composites, maintaining high levels of photocatalytic hydrogen production over eight cycles without significant efficiency loss, indicating their impressive durability. This investigation presents a promising strategy for crafting and producing environmentally sustainable SnO2@ZnIn2S4 composites with prospective implementations in photocatalytic hydrogen generation.

Plastic and Micro/Nanoplastic Pollution in Sub-Saharan Africa: Challenges, Impacts, and Solutions

Sub-Saharan Africa faces increasing levels of plastic production and importation, unregulated usage, and inadequate waste management systems. This region’s harsh conditions often lead to plastic breaking down into microplastics and nanoplastics. This review explores the abundance of micro/nanoplastics across different environmental mediums, such as surface waters, sediments, and aquatic organisms, in sub-Saharan African countries. It also highlights knowledge gaps concerning the region’s abundance of micro/nanoplastics. The effects of plastics and micro/nanoplastics on food production, water quality, health, and the environment are discussed. Strategies to address the challenges of plastic pollution are proposed. Finally, the review concludes with future perspectives for addressing the ongoing challenges of plastic waste management in sub-Saharan Africa. The materials for this study were sourced from published articles on Scopus, Google Scholar, ResearchGate, and additional platforms, including reports and various press releases, using keywords such as plastic waste, micro/nano-plastic, sub-Saharan Africa, toxicity, and circular economy. Articles were initially screened by reviewing abstracts, followed by a thorough reading of full papers to identify relevant studies. Key information was extracted from these selected articles and incorporated into this review.

Increasing the efficiency of gene editing with CRISPR-Cas9 via concurrent expression of the Beta protein

Escherichia coli has emerged as an important host for the production of biopharmaceuticals or other industrially relevant molecules. An efficient gene editing tool is indispensable for ensuring high production levels and optimal release of target products. However, in Escherichia coli, the CRISPR-Cas9 system has been shown to achieve gene modifications with relatively low frequency. Large-scale PCR screening is required, hindering the identification of positive clones. The beta protein, which weakly binds to single-stranded DNA but tightly associates with complementary strand annealing products, offers a promising solution to this issue. In the present study, we describe a targeted and continuous gene editing strategy for the Escherichia coli genome. This strategy involves the coexpression of the beta protein alongside the CRISPR-Cas9 system, enabling a variety of genome modifications such as gene deletion and insertion with an efficiency exceeding 80 %. The integrity of beta proteins is essential for the CRISPR-Cas9/Beta-based gene editing system. In this work, the deletion of either the N- or C-terminal domain significantly impaired system efficiency. Overall, our findings established the CRISPR-Cas9/Beta system as a suitable gene editing tool for various applications in Escherichia coli.

Process optimization for the production of fibrinolytic enzyme by a novel marine fungus – Penicillium steckii KU1

The demand for microbial thrombolytic agents is alarmingly increasing, and the discovery of novel microbial species with significant level of fibrinolytic enzyme production are required to cope up with the current circumstances. Surprisingly, marine fungi remain largely unexplored in this regard, presenting an untapped potential for the production of fibrinolytic enzymes.A novel marine fungal strain (Penicillium steckii KU1) was isolated successfully from Kappil beach, Kerala. At the beginning Czapek-dox media was used for the production of fibrinolytic enzyme. Statistical optimization of the enzyme production was achieved initially by Plackett-Burman design (PB). Interactive effect of various process parameters was further scrutinized by Central Composite Design (CCD). Incubation time, inoculum size and concentration of casein were shown significant contribution towards production. Statistical optimization, resulted in a tenfold increment (454.49 ± 14.66 U/mL) [optimized conditions - 30 °C, pH 7, casein 2.7 % (w/v), inoculum 5.75 million spores/mL and 115 h incubation], against the predicted value 441.42 U/mL.

Inhibitory mechanism of carboxymethyl cellulose on advanced glycation end products in plant-based meat alternatives

Plant-based meat alternatives (PBMA) were detected with high advanced glycation end products (AGEs) levels in our previous study, threatening human health. In this study, carboxymethyl cellulose (CMC) exhibited excellent inhibitory effects on AGEs in both PBMA and chemical model systems. Moreover, the inhibitory effects were dependent on the levels of CMC addition. With 4% CMC addition, the contents of Nε-carboxymethy-l-lysine (CML) and Nε-carboxyethy-l-lysine (CEL) in PBMA decreased by 75% and 83%, respectively. Through comprehensive characterizations and high-resolution mass spectrometry analysis, three possible inhibitory pathways were proposed at the molecular level: CMC improved the thermal stability of soy protein isolates (SPI) via covalent bond; CMC occupied the amino reaction sites on SPI and competitively inhibited the reaction between SPI and glucose; CMC scavenged the intermediates of AGEs (i.e glyoxal and methylglyoxal). This research lays a theoretical foundation on the inhibition of AGEs in PBMA through the regulation of hydrocolloids.

Middle Ordovician climatic and oceanic destabilization in a slope-setting of the Yangtze platform, South China, and its role as a regional brake on the Ordovician radiations

During the Middle Ordovician, the Great Ordovician Biodiversification Event (GOBE) saw an accelerated diversification pulse in the marine metazoan richness, which has been linked to the oceanic and climatic changes. Here we have examined a deep-water slope succession, Xijinhe section, in Lower Yangtze region by integrating sedimentological and geochemical methods. This study highlights that the climatic and oceanic destabilization might have substantially impacted the marine biodiversity during the Middle Ordovician in South China. Three phases are identified based on lithological and geochemical variations. During Phase I across the Early–Middle Ordovician transition (Floian–Dapingian), the TOC and abundances of redox sensitive elements (RSE) are relatively depleted, suggesting a suboxic-oxic environment in the slope area of Yangtze Sea. The continental chemical weathering intensity transits from stable to unstable in Phase I indicated by CIA (chemical index of alteration) that gradually fluctuate to lower values, potentially caused by the secular cooling. The slowly ameliorating climate and redox conditions advanced the biodiversification across the Early–Middle Ordovician transition. During the Phase II in the Darriwilian, the continental chemical weathering intensity continues to decrease and the concentrations of RSE start to rise. The values of Mn(%)*Co(ppm) become consistently lower than 0.4, and the bio-limited elements such as P, Ba, and Ni are substantially enriched correspondingly. This phenomenon implicates that a secular cooling climate would have resulted in oceanic destabilization by improving upwelling of deeper anoxic/euxinic and eutrophic water into epicontinental sea, potentially causing a deceleration in the biodiversification during the Darriwilian in South China. The Phase III across the Middle–Late Ordovician transition, witnesses the continuous decrease of CIA values and highest abundances of TOC and RSE though some minor fluctuations, indicating a possibly even more drastic oceanic destabilization relative to Phase II. Deeper anoxic/euxinic water continuously intruded and shoaled into the shelf area during the Phase III due to the enhanced upwelling. Consequently, the habitat of marine organisms in shallow water was contracted, and becoming harsher. Additionally, bio-limited elements such as P were released from anoxic sediments and recycle into the seawater during the Middle Ordovician, boosting primary productivity level and may further deplete oxygen in seawater. This unstable oceanic state during the Middle Ordovician, particularly Darriwilian, represented by high nutrients content in epi-continental sea and expanding anoxic/euxinic water, could be attributable for the deceleration of biodiversification in South China and potentially the spatially variable paces of biodiversification in different paleo-continents during the main phase of the GOBE.

Changing the rules of the game: how firms respond to the revision of responsible product standards

Abstract While voluntary responsible business standards are more flexible than regulation and thus more responsive to changes in society, revising standards can have negative effects on compliance rates. We hypothesize that when non-compliance is visible to external stakeholders and participating firms receive a clear signal that their products are non-compliant, the firms will respond by re-certifying their products, which will raise their certification rates. We tested our hypothesis by conducting a multi-level regression on the certification rates before and after the revision of a voluntary label for healthier food products, active in the Dutch market from 2006 until 2015 in the Dutch market (76 firms, 22 product categories, 1354 firm-category-year observations). Our results show the opposite effect than hypothesized: after standards’ revision, firms with decertified products significantly decrease their certification rates compared to firms without decertified products. We conclude that making standards stricter over time is not always a successful strategy to maintain compliance. We argue that this decrease in compliance after standards’ revisions can be explained by the low behavioural visibility of non-compliance, restricting stakeholder pressure both at the product level by consumers as well as at the firm level by other stakeholders.

A Study on Capacity Planning in Pharmaceutical Manufacturing: with Reference to Select Pharma Companies in India

Capacity planning is the method and planning of how efficiently the company can utilize its resources to meet the market demand. A well-executed capacity planning helps the organization to avoid the stock out conditions and serve the customers without any failure. The pharmaceutical capacity planning is an important aspect as this sector directly deals with mankind. The field of study is significant in order to meet the future demands, improve resource allocation, quality of the product and service level. This research work includes the secondary data collection through industrial reports, financial reports, case studies and various research papers. The main factors included for the research are the R&D investments, plant & equipment costs and inventory cost as these factors directly influence the capacity planning in manufacturing. The key findings of this study concludes that the pharma industries are continuously focusing on improving the efficiency in manufacturing by adopting the technology, reducing cost and expanding market size.