Moorpark College

The development of molecular cloning techniques starting in the late 1960s created new tools to facilitate our understanding of gene function. Later advances in PCR and site-specific mutagenesis allowed researchers to efficiently dissect genes down to the single-nucleotide level. These methods have become virtually ubiquitous in nearly every biology laboratory. Recently, we developed an additional technique called SIMPLE (SapI/AarI incision-mediated plasmid editing), which allows for efficient deletions, short insertions (e.g., epitope tagging), and accurate mutations of episomal plasmids. SIMPLE cloning has a wide range of applications for molecular geneticists and adds a precision instrument to the biologist's toolbox. Here, we describe a detailed, step-by-step set of instructions for SIMPLE cloning.

Background: The LIPA gene encodes for lysosomal acid lipase (LAL), which catalyzes the hydrolysis of cholesterol esters and triglycerides. Variations in the LIPA gene impair LAL activity, predisposing patients to a rare metabolic disorder called LAL deficiency (LAL-D). The lack of functioning LAL promotes lipid accumulation and subsequent dyslipidemia, which can increase the likelihood of complications in both infants and adults. Although the worldwide prevalence is 1:500,000 births, the frequency in Mizrahi Jewish populations is projected to be as high as 1 in every 4200 births (Valles-Ayoub et al.) based on the LIPA p.G87V variant frequency among 162 individuals. Materials and Methods: This study was conducted to validate the previously reported prevalence of LAL-D in the Mizrahi Jewish population based on the pathogenic LIPA missense variants in exon 4 (c.260G>T; p.G87V) and exon 8 (c.894G>A; p.Gln298=) using a larger cohort of those with Middle Eastern ancestry living around Los Angeles. Among the 1184 individual samples sequenced, 660 self-reported as Mizrahi Jewish, while the remaining 524 came from other Middle Eastern groups labeled as "non-Jewish." Results: Of the 1184 samples, 22 alleles of the exon 4 variant were identified (1.85%), and 2 alleles of the exon 8 variant were identified (0.16%). For the exon 4 variant, 20 of 22 (90.9%) heterozygotes were Mizrahi Jewish, while 2 of 22 (9.09%) heterozygotes were "non-Jewish." For the exon 8 variant, 2 of 2 (100%) heterozygotes were Mizrahi Jewish. This suggests that the prevalence of LAL-D in this population is 1 in 900, which suggests that LAL-D may be 4.6% higher in the Mizrahi Jewish population in previous reports. Conclusion: These findings show increased prevalence of LIPA gene exon 4 variation p.G87V in the Middle East population when compared to the general population, indicating the need for prenatal screening in those of Mizrahi Jewish ancestry.

Pore size, external shape, and internal complexity of additively manufactured porous titanium scaffolds are three primary determinants of cell viability and structural strength of scaffolds in bone tissue engineering. To obtain an optimal design with the combination of all three determinants, four scaffolds each with a unique topology (external geometry and internal structure) were designed and varied the pore sizes of each scaffold 3 times. For each topology, scaffolds with pore sizes of 300, 400, and 500 µm were designed. All designed scaffolds were additively manufactured in material Ti6Al4V by the direct metal laser melting machine. Compression test was conducted on the scaffolds to assure meeting minimum compressive strength of human bone. The effects of pore size and topology on the cell viability of the scaffolds were analyzed. The 12 scaffolds were ultrasonically cleaned and seeded with NIH3T3 cells. Each scaffold was seeded with 1 million cells. After 32 days of culturing, the cells were fixed for their three-dimensional architecture preservation and to obtain scanning electron microscope images.

For points p1,…,pn in the unit square [0,1]2, an anchored rectangle packing consists of interior-disjoint axis-aligned empty rectangles r1,…,rn⊆[0,1]2 such that point pi is a corner of the rectangle ri (that is, ri is anchored at pi) for i=1,…,n. We show that for every set of n points in [0,1]2, there is an anchored rectangle packing of area at least 7∕12−O(1∕n), and for every n∈N, there are point sets for which the area of every anchored rectangle packing is at most 2∕3. The maximum area of an anchored square packing is always at least 5∕32 and sometimes at most 7∕27.The above constructive lower bounds immediately yield constant-factor approximations, of 7∕12−ε for rectangles and 5∕32 for squares, for computing anchored packings of maximum area in O(nlogn) time. We prove that a simple greedy strategy achieves a 9∕47-approximation for anchored square packings, and 1∕3 for lower-left anchored square packings. Reductions to maximum weight independent set (MWIS) yield a QPTAS for anchored rectangle packings in exp(poly(ε−1logn)) time and a PTAS for anchored square packings in nO(1∕ε) time.

Innovation and leadership are so closely interrelated that leadership thinks of the future and innovation is the fruit of leadership in the future. This paper aims to determine whether leadership leads to innovation. Under visionary leadership, innovation can progress and bear fruit. An appropriate framework to provide a platform for innovation is a necessary condition and requires the appropriate leadership to guide its flow in that direction. Competency for innovation arrises from two main cores: interior and exterior. The exterior core has three components: culture, resources, and habitat; the interior core also has three components: knowledge, attitude, and imagination. The components of, the exterior core, relates to the outside world and one’s surroundings whereas the components of the interior core relate to us as individuals. The job of leadership is to create a platform wherein the inner forces synchonize with the outside forces. This paper uses Seelig’s Innovation Engine, model to measure innovation progress in multiple industries in the Gulf region and to identify the most innovative industries. A survey, distributed to leaders in several major industries, was collected, analyzed, and presented.

The cost of testing (COT) has been a prominent issue for clinical cytogenetic laboratory directors and managers in the last few years. There is little standardization of the costs of chromosome, FISH, or microarray testing. This chapter addresses common practices among laboratories, taking into account staff costs, supplies, time assessment of laboratory tasks, and overhead, as well as direct and indirect costs in a laboratory. Direct costs include labor and cost of materials to perform tests, including the cost of supplies needed for specimen processing, and equipment, such as microscopes, incubators, refrigerators, and freezers. Indirect costs include labor of individuals that are not in the direct process of testing but are needed to assist, such as administrative staff and managers and supervisors that do not perform testing but only supervise others. Good laboratory practice usually is defined by an average of 70–80% direct time of testing personnel with the remainder as indirect time.

Abnormal Any change from the "correct" or "usual."It may not necessarily mean harmful or undesirable; it can equally mean atypical, unusual, or uncommon.When used in reference to genes, an abnormal gene may result in a genetic condition.Accession The process of identifying a specimen and entering a unique specimen identifier into laboratory records.Accuracy A measure of how close a measured value is to the true value.ACGIH The American Conference of Governmental Industrial Hygienists is a voluntary membership organization of professional industrial hygiene personnel in governmental or educational institutions.The ACGIH develops and publishes recommended occupational exposure limits each year called threshold limit values (TLV's) for hundreds of chemicals, physical agents, and biological exposure indices.Acrocentric Acrocentric chromosomes are those with the centromere very close to one end, giving the chromosome a "V"-shaped appearance.The acrocentric chromosomes are numbers 13, 14, 15, 21, and 22.The short "p" arms are very short and usually have small round appendages on stalks, known as "satellites."Action level A concentration designated in 29 CFR Part 1910 for a specific substance, calculated as an eight ( 8)-hour time-weighted average (TWA), that initiates certain required activities such as exposure monitoring and medical surveillance.Acute effect Symptom of exposure to a hazardous material that soon appears after a shortterm exposure, coming quickly to a crisis.Acute exposure: a single, brief exposure to a large dose of a toxic substance.Adverse health effects are evident soon after exposure.Acute toxicity: an adverse biological effect caused by a single dose of a toxic agent.Aerosol: a suspension of fine solid or liquid particles in that resides in the air (e.g., paint spray, mist, and fog). GlossAryGLoSSARy Adenine (A) one of the four basic building blocks (nucleotide bases) that makes up the genetic code or DNA.It is represented by the letter A. Adult stem cell An undifferentiated cell found in a differentiated tissue that can renew itself and (with certain limitations) differentiate to yield all the specialised cell types of the tissue from which it originated.Alert values See "Critical values."Aliquot A portion of a specimen or product used for testing.Aliquoting The action of dispensing a product or specimen into smaller quantities.A portion of this product or specimen (aliquot) is typically used for testing or placed into long-term storage.Allele There are usually two copies of a gene.These two copies are called "alleles."In some cases, one or both alleles will be mutated or altered in some way.American National standards Institute (ANsI) organization that oversees the creation, proliferation, and use of thousands of rules and guidelines that affect a broad range of businesses; also engaged in accrediting programs that assess conformance to standards, such as ISo 9000 management systems (http://www.ansi.org).This privately funded, voluntary organization also develops standards for the safe design and operation of equipment and safe practices or procedures for industry.American society for Testing and Materials (AsTM) organization that develops and generates voluntary standards that are used worldwide (http://www.astm.org).Amino acids Small chemical building blocks that join together to form proteins: there are 20 common amino acids that join in different combinations to make up proteins.Amniocentesis A procedure for obtaining amniotic fluid for prenatal diagnosis.Using a sterile needle, a sample of amniotic fluid is removed from the uterus; the amniotic fluid contains cells from the fetus that can be analyzed to determine if the fetus has a specific condition.The test may be carried out from the 15th week of pregnancy.Amniotic fluid Fluid in which the fetus floats in the uterus; fetal cells are found suspended in this fluid.Analyte The substance being measured in an analytical procedure.Analytical measurement range (AMr) The range of analyte values that a method can directly measure on the specimen (without any pretreatment not included as a procedural step of the assay).Analytical run An interval, period of time, or number of specimen for which the precision and accuracy of the method is expected to remain stable.Analytical sensitivity The estimate of the lowest concentration of an analyte that can be measured. Analytical specificity (analytical interferences)The estimate of the systematic error caused by other materials that may be present in the specimen being analyzed (e.g., lipemia and drugs).Anesthetic A chemical that causes drowsiness.Large doses of anesthetic chemicals can cause unconsciousness, coma, and death.Aneuploidy The gain or loss of chromosomal material, that is, having missing or extra chromosomes.Antibody A protein, produced in response to a foreign substance in the immune system.

A quality management (QM) program is essential to ensure the safety of patient samples and maintenance of quality laboratory operations. The QM program should include a quality assurance (QA) program, which is set up to evaluate the laboratory's analytical performance by comparing test performance. The College of American pathologists (CAP) has specific guidelines on developing and implementing QM programs and their component plans. Individual assay controls must be in place to ensure assay performance. The laboratory must have written standard operating procedures (SOPs) for laboratory processes to ensure the quality, and integrity of the test data. If specific water types are required per manufacturer for certain testing procedures, the laboratory must ensure that records of water quality testing are complete and indicate that the required standards for water quality are consistently met. Reports generated by a laboratory information system (LIS) must be concise, readable, and in a standardized format.

Cell culture processes, the in vitro growth of animal, insect, or plant cells on a large scale to manufacture biochemicals or cells for therapeutic purposes, have evolved and grown significantly over the past 10 years. Significant growth in this technology, primarily because of the advent of recombinant DNA methods for the production of therapeutic proteins and hybridoma technology for the production of monoclonal antibodies, occurred in the late 1980s and 1990s. During the past decade, many developments have led to increased process efficiencies and much higher product titers and made this technology ubiquitous in the biopharmaceutical industry. Today, worldwide products of cell culture technology command revenues of more than $60 billion per year. Additionally, the advent of stem cells and regenerative medicine has ushered in the need for novel methods of cell culture that are propelling the industry in new directions such as the use of single‐use bioreactors for personalized medicine. Single‐use bioreactor technology is also finding use in large‐scale production of protein therapeutics. This article provides information on the various processes and equipment used in the production of monoclonal antibodies (MAbs), recombinant therapeutics, vaccines, and other products.

We present a statistical analysis of the millimeter-wavelength properties of 1.4 GHz-selected sources and a detection of the Sunyaev-Zel'dovich (SZ) effect associated with the halos that host them. The Atacama Cosmology Telescope (ACT) has conducted a survey at 148 GHz, 218 GHz and 277 GHz along the celestial equator. Using samples of radio sources selected at 1.4 GHz from FIRST and NVSS, we measure the stacked 148, 218 and 277 GHz flux densities for sources with 1.4 GHz flux densities ranging from 5 to 200 mJy. At these flux densities, the radio source population is dominated by active galactic nuclei (AGN), with both steep and flat spectrum populations, which have combined radio-to-millimeter spectral indices ranging from 0.5 to 0.95, reflecting the prevalence of steep spectrum sources at high flux densities and the presence of flat spectrum sources at lower flux densities. The thermal SZ effect associated with the halos that host the AGN is detected at the 5$\sigma$ level through its spectral signature. When we compare the SZ effect with weak lensing measurements of radio galaxies, we find that the relation between the two is consistent with that measured by Planck for local bright galaxies. We present a detection of the SZ effect in some of the lowest mass halos (average $M_{200}\approx10^{13}$M$_{\odot}h_{70}^{-1}$) studied to date. This detection is particularly important in the context of galaxy evolution models, as it confirms that galaxies with radio AGN also typically support hot gaseous halos. With Herschel observations, we show that the SZ detection is not significantly contaminated by dust. We show that 5 mJy$<S_{1.4}<$200 mJy radio sources contribute $\ell(\ell+1)C_{\ell}/(2\pi)=0.37\pm0.03\mu$K$^2$ to the angular power spectrum at $\ell=3000$ at 148 GHz, after accounting for the SZ effect associated with their host halos.