β-Actin—an unsuitable internal control for RT-PCR

Despite reports confirming cell-cycle dependent gene expression and a number of studies describing specific circumstances in which β-actin is also regulated, the mRNA for β-actin remains a widely used housekeeping gene internal control. Utilizing differential reverse transcriptase-polymerase chain reaction (RT-PCR), we report here the dose-dependent inhibition of β-actin by matrigel. This was detected by comparison to the very moderate inhibition of the target gene, membrane type-1 matrix metalloproteinase (MT1-MMP), with results independently confirmed by similar findings on MT1-MMP expression using competitive RT-PCR. Furthermore, RT-PCR of the housekeeping gene 18 Svedberg Units (S) rRNA demonstrated excellent consistency, reproducibility and non-regulation by a matrigel treatment. We conclude that β-actin is highly regulated by matrigel and therefore unsuitable as an internal control in this treatment. Hence, these findings suggest that researchers have a responsibility to ensure that the housekeeping gene of choice is not regulated in their specific application, as such regulation may dramatically affect the accuracy of their results. This study reinforces the necessity for minimally regulated housekeeping genes such as 18S rRNA, and the superiority of competitive templates as internal controls for quantitative applications of RT-PCR.

Effects Of 5' Flanking Sequences And Changes In The 5' Internal Control Region On The Transcription Of Rice Transfer-Rna Gcc Cly Gene

A stretch of 71 nucleotides in a 1.2 kilobase pair Pst I fragment of rice DNA was identified as tRNA~ gene by hybridization and nucleotide sequence analyses. The hybridization of genomic DNA with the tRNA gene showed that there are about 10 glycine tRNA genes per diploid rice genome. The 3' and 5' internal control regions, where RNA polymerase III and transcription factors bind, were found to be present in the coding sequence. The gene was transcribed into a 4S product in an yeast cell-free extract. The substitution of 5' internal control region with analogous sequences from either M13mpl9 or M13mpl8 DNA did not affect the transcription of the gene in vitro. The changes in three highly conserved nucleotides in the consensus 5' internal control region (RGYNNARYGG; R = purine, Y = pyrimidine, N = any nucleotide) did not affect transcription showing that these nucleotides are not essential for promotion of transcription. There were two 16 base pair repeats, 'TGTTTGTTTCAGCTTA' at - 130 and - 375 positions upstream from the start of the gene. Deletion of 5' flanking sequences including the 16 base pair repeat at - 375 showed increased transcription indicating that these sequences negatively modulate the expression of the gene.

Smallholder group certification in Uganda - Analysis of internal control systems in two organic export companies

The organic agricultural sector of Uganda is among the most developed in Africa in terms of its professional institutional network and high growth rates of number of certified farmers and land area. Smallholder farmers are certified organic through contract production for export companies using a group certification scheme (internal control system - ICS). The ICS is a viable and well-accepted tool to certify small-scale producers in developing countries all over the world. Difficulties in certification are still stated to be among the main constraints for Uganda’s organic sector development. Therefore, this paper reports a qualitative case study comprising 34 expert interviews in two organic fresh-produce export companies in central Uganda, aiming to explore the challenges which underlie organic certification with ICS. The study shows that farmers cannot be labelled as ‘organic by default’ but deliberately engage in organic production as a marketing strategy. The small quantities purchased by the organic companies lead to a difficult marketing situation for the farmers, causing production and infiltration risks on the farm level. These risks require increased control that challenges the companies organizationally. The risks and control needs are a reason to involve farmers in ICS procedures and innovatively adapt the ICS by means of a bypass around formal perspective restrictions. The paper discusses different perspectives on risks, risk control and certification.

Identification of suitable internal control genes for expression studies in Coffea arabica under different experimental conditions

Background: Quantitative data from gene expression experiments are often normalized by transcription levels of reference or housekeeping genes. An inherent assumption for their use is that the expression of these genes is highly uniform in living organisms during various phases of development, in different cell types and under diverse environmental conditions. To date, the validation of reference genes in plants has received very little attention and suitable reference genes have not been defined for a great number of crop species including Coffea arabica. The aim of the research reported herein was to compare the relative expression of a set of potential reference genes across different types of tissue/organ samples of coffee. We also validated the expression profiles of the selected reference genes at various stages of development and under a specific biotic stress.Results: The expression levels of five frequently used housekeeping genes (reference genes), namely alcohol dehydrogenase (adh), 14-3-3, polyubiquitin (poly), beta-actin (actin) and glyceraldehyde-3-phosphate dehydrogenase (gapdh) was assessed by quantitative real-time RT-PCR over a set of five tissue/organ samples (root, stem, leaf, flower, and fruits) of Coffea arabica plants. In addition to these commonly used internal controls, three other genes encoding a cysteine proteinase (cys), a caffeine synthase (ccs) and the 60S ribosomal protein L7 (rpl7) were also tested. Their stability and suitability as reference genes were validated by geNorm, NormFinder and BestKeeper programs. The obtained results revealed significantly variable expression levels of all reference genes analyzed, with the exception of gapdh, which showed no significant changes in expression among the investigated experimental conditions.Conclusion: Our data suggests that the expression of housekeeping genes is not completely stable in coffee. Based on our results, gapdh, followed by 14-3-3 and rpl7 were found to be homogeneously expressed and are therefore adequate for normalization purposes, showing equivalent transcript levels in different tissue/ organ samples. Gapdh is therefore the recommended reference gene for measuring gene expression in Coffea arabica. Its use will enable more accurate and reliable normalization of tissue/organ-specific gene expression studies in this important cherry crop plant.

Resistance to Temptation: The Interaction of External and Internal Control on Alcohol Use During Residential Treatment for Alcohol Use Disorder

Background: There is evidence that drinking during residential treatment is related to various factors, such as patients’ general control beliefs and self-efficacy, as well as to external control of alcohol use by program’s staff and situations where there is temptation to drink. As alcohol use during treatment has been shown to be associated with the resumption of alcohol use after discharge from residential treatment, we aimed to investigate how these variables are related to alcohol use during abstinenceoriented residential treatment programs for alcohol use disorders (AUD). Methods: In total, 509 patients who entered 1 of 2 residential abstinence-oriented treatment programs for AUD were included in the study. After detoxification, patients completed a standardized diagnostic procedure including interviews and questionnaires. Drinking was assessed by patients’ selfreport of at least 1 standard drink or by positive breathalyzer testing. The 2 residential programs were categorized as high or low control according to the average number of tests per patient. Results: Regression analysis revealed a significant interaction effect between internal and external control suggesting that patients with high internal locus of control and high frequency of control by staff demonstrated the least alcohol use during treatment (16.7%) while patients with low internal locus of control in programs with low external control were more likely to use alcohol during Treatment (45.9%). No effects were found for self-efficacy and temptation. Conclusions: As alcohol use during treatment is most likely associated with poor treatment outcomes, external control may improve treatment outcomes and particularly support patients with low internal locus of control, who show the highest risk for alcohol use during treatment. High external control may complement high internal control to improve alcohol use prevention while in treatment. Key Words: Alcohol Dependence, Alcohol Use, Locus of Control, Alcohol Testing.

Streamlining internal control processes and strengthening management controls with less effort /

Item 851-J

Independent auditors' report on the internal control structure /

Description based on: Year ended Sept. 30, 1994; title from cover.

Internal control course.

Cover title.

Quality improvement through the computerized internal control review (CICR) system.

Shipping list no.: 89-358-P.

International risk management:systems, internal control and corporate governance

This book is very practical in its international usefulness (because current risk practice and understanding is not equal across international boundaries). For example, an accountant in Belgium would want to know what the governance regulations are in that country and what the risk issues are that he/she needs to be aware of. This book covers the international aspect of risk management systems, risk and governance, and risk and accounting. In doing so the book covers topics such as: internal control and corporate governance; risk management systems; integrating risk into performance management systems; risk and audit; governance structures; risk management of pensions; pension scheme risks e.g. hedging derivatives, longevity bonds etc; risk reporting; and the role of the accountant in risk management. There are the case studies through out the book which illustrate by way of concrete practical examples the major themes contained in the book. The book includes highly topical areas such as the Sarbanes Oxley Act and pension risk management.

Internal control reporting by non-accelerated filers

I examine three issues related to internal control reporting by non-accelerated filers. Motivation for the three studies comes from the fact that Section 404 of the Sarbanes-Oxley Act (SOX) continues to be controversial, as evidenced by the permanent exemption from Section 404(b) of SOX granted to non-accelerated filers by the Dodd-Frank Wall Street Reform and Consumer Protection Act of 2010. The Dodd-Frank Act also requires the SEC to study compliance costs associated with smaller accelerated filers. In the first part of my dissertation, I document that the audit fee premium for non-accelerated filers disclosing a material weakness in internal controls (a) is significantly lower than the corresponding premium for accelerated filers, and (b) declines significantly over time. I also find that in the case of accelerated filers remediating clients pay lower fees compared to clients continuing to report internal control problems; however, such differences are not observed in the case of non-accelerated filers. The second essay focuses on audit report lag. The results indicate that presence of material weaknesses are associated with increased audit report lags, for both accelerated and non-accelerated filers. The results also indicate that the decline in report lag following remediation of problems is greater for accelerated filers than for non-accelerated filers. The third essay examines early warnings (pursuant to Section 302 disclosures) for firms that subsequently disclosed internal control problems in their 404 reports. The analyses indicate that non-accelerated firms with shorter CFO tenure, presence of accounting experts on the audit committee, and more frequent audit committee meetings are more likely to provide prior Section 302 warnings. Overall the results suggest that there are differences in internal control reporting between the accelerated and non-accelerated filers. The results provide empirical grounding for the ongoing debate about internal control reporting by non-accelerated filers.

Internal Control application : Empirical evidence from Sweden

Aim The aim of this study is to explore based on internationally recognised frameworks: 1. how internal control structures are applied in Sweden among different sectors; 2. how organizational size and environment affect internal control structures; and 3. the impact of internal control structures on organizational performance. Methods A quantitative method was used in the data collection and analysis. The sample consisted of 1117 organizations operating in Sweden. A mean analysis was conducted to measure the level of internal control structures among different industries, organizational sizes, and different choices of listing in the stock exchange market. Person’s correlation analysis was then used to explore possible correlations between external environmental factors and internal control structures, and internal control structures and organizational performance. Lastly, a structural model was built to measure the impact of internal control structures on organizational performance. The measurements of internal control structures and organizational performance are based on COSO framework’s principles and objectives. Results This study gives an insight on how internal control structures are applied across industrial sectors in Sweden, with financial institutions and manufacturing organizations having notably higher levels of internal control structures. Additionally, it provides evidence of the impact external environmental factors have on internal control structures. Furthermore, it shows that organizations that are listed in the Swedish stock exchange market have an equivalent level of internal control structures to those registered in the American stock exchange market. In contrast, organisations that are not listed in the stock exchange market have a notably lower level of internal control structures. Lastly, it illustrates the positive impact the presence of internal control structures has on organizational performance. 3 | P a g e Conclusion The results highlight a crucial role the supervisory authority Finansinspektionen (FI) has in regulating the Swedish financial market. They also show that the stability of the Swedish business environment has had a positive impact on the level of internal control structures.

Internal audit department characteristics/activities and audit fees : some evidence from Hong Kong firms

This study provides preliminary support for the notion that internal audit function assists in reducing external audit effort and fees. Data on internal audit characteristics and activities are obtained from survey respondents of Hong Kong companies and audit fee model data are acquired from their annual reports. The results of this study suggest that the external auditor of firms in Hong Kong rely on the internal audit function and subsequently charge a lower fee. Lower external audit fees are associated with a larger internal audit department and certain activities carried out by the internal audit. Specifically, lower external audit fees are associated with more internal audit effort spent on activities relating to financial statements, systems development and maintenance, operating efficiency and effectiveness, fraud investigations and unlimited access to internal auditors’ working papers. The results of this study suggest that the contribution of the internal audit may substitute for some substantive external auditing processes and lower monitoring costs.

Adaptive compliance control of a multi‐legged stair‐climbing robot based on proprioceptive data

Purpose – The purpose of this paper is to describe an innovative compliance control architecture for hybrid multi‐legged robots. The approach was verified on the hybrid legged‐wheeled robot ASGUARD, which was inspired by quadruped animals. The adaptive compliance controller allows the system to cope with a variety of stairs, very rough terrain, and is also able to move with high velocity on flat ground without changing the control parameters. Design/methodology/approach – The paper shows how this adaptivity results in a versatile controller for hybrid legged‐wheeled robots. For the locomotion control we use an adaptive model of motion pattern generators. The control approach takes into account the proprioceptive information of the torques, which are applied on the legs. The controller itself is embedded on a FPGA‐based, custom designed motor control board. An additional proprioceptive inclination feedback is used to make the same controller more robust in terms of stair‐climbing capabilities. Findings – The robot is well suited for disaster mitigation as well as for urban search and rescue missions, where it is often necessary to place sensors or cameras into dangerous or inaccessible areas to get a better situation awareness for the rescue personnel, before they enter a possibly dangerous area. A rugged, waterproof and dust‐proof corpus and the ability to swim are additional features of the robot. Originality/value – Contrary to existing approaches, a pre‐defined walking pattern for stair‐climbing was not used, but an adaptive approach based only on internal sensor information. In contrast to many other walking pattern based robots, the direct proprioceptive feedback was used in order to modify the internal control loop, thus adapting the compliance of each leg on‐line.