Plasmid DNA is prepared from the recombinant shuttle vector pLD53.SCAB/A-B created by cloning of the A and B homology arms for two-step bacterial artificial chromosome (BAC) engineering. To confirm that the A-box and B-box arms have been successfully incorporated into pLD53.SCAB, the pattern of enzyme digestion of the modified plasmid is compared with that of the unmodified pLD53.SCAB. Once the shuttle vector is shown to carry the proper sequences, it is ready for transfer into the BAC host.

The 700-bp A homology arm (A-box) and the 700-bp B homology arm (B-box) are amplified by polymerase chain reaction (PCR) using purified bacterial artificial chromosome (BAC) DNA as template for two-step BAC engineering. The resulting A-box PCR product contains an AscI site at its 5' end (the 5' primer incorporates an AscI site, and the 3' primer does not incorporate any restriction sites). The B-box PCR product contains an XmaI site at its 3' end (the 5' primer does not incorporate any restriction sites, and the 3' primer incorporates an XmaI site). The amplification products are then digested with the appropriate restriction endonucleases to render them suitable for cloning into the shuttle vector.

In two-step bacterial artificial chromosome (BAC) engineering, a single plasmid is introduced into the BAC-carrying cell lines. The shuttle vector pLD53.SCAB (or pLD53.SCAEB) carries the recA gene and the R6K origin, which requires the protein to replicate. PIR2 cells, expressing , are typically used for the amplification of the vector and maintain about 15 copies/cell of the donor vector, which is relatively stable in this host.

Schools in western Quebec are getting ready to welcome students back next week, but things will look a lot different — and classrooms will likely be far from full. 

Toronto officials reported 180 new cases of COVID-19 on Thursday, a figure that represents a dip when compared to the more than 200 cases announced daily this week. 

Living on a 10-acre plot of land 100 kilometres outside Toronto, David Stevens thought he and his family were well prepared to ride out the COVID-19 quarantine. But after a call from his mother, he learned that even the best laid plans can go wrong.

Deforestation in Brazil's Amazon rainforest rose sharply in April, government data showed on Friday, as the coronavirus outbreak keeps many environmental enforcers out of the field and the country prepares to deploy troops to fight illegal logging.

A normal draft year would see NHL scouts and executives spend the spring crisscrossing Europe and North America collecting information. The circumstances surrounding a 2020 draft and the COVID-19 pandemic, however, aren't anything resembling normal.

Rome Newsroom, Apr 30, 2020 / 11:45 am (CNA).- After eight weeks without funerals, Italian families will be able finally to gather together to mourn and pray at funeral Masses for the victims of the coronavirus starting May 4.

In Milan, the largest city in Italy’s coronavirus epicenter, priests are preparing for an influx of funeral requests in the coming weeks in the Lombardy region, where 13,679 have died.

Fr. Mario Antonelli, who oversees liturgies on behalf of the Archdiocese of Milan, told CNA that archdiocesan leadership met April 30 to coordinate guidelines for Catholic funerals as more than 36,000 people remain positive for COVID-19 in their region.

“I am moved, thinking of so many dear people who have wanted [a funeral] and still desire one,” Fr. Antonelli said April 30.

He said that the church in Milan is ready like the Good Samaritan to “pour oil and wine on the wounds of many who have suffered the death of a loved one with the terrible agony of not being able to say goodbye and embrace.”

A Catholic funeral is “not just a solemn farewell from loved ones,” the priest explained, adding that it expresses a pain like childbirth. “It is the cry of pain and loneliness that becomes a song of hope and communion with the desire for an everlasting love.”

Funerals in Milan will occur on an individual basis with no more than 15 people in attendance, as required by “phase two” of the Italian government’s coronavirus measures. 

Priests are asked to notify local authorities when a funeral is scheduled to take place and ensure that social distancing measures defined by the diocese are followed throughout the liturgy. 

Milan is home to the Ambrosian rite, the Catholic liturgical rite named for St. Ambrose, who led the diocese in the 4th century.

“According to the Ambrosian rite, the funeral liturgy includes three ‘stations’: the visit / blessing of the body with the family; community celebration (with or without Mass); and burial rites at the cemetery,” Antonelli explained. 

“Trying to reconcile the sense of the liturgy … and the sense of civic responsibility, we ask the priests to refrain from visiting the family of the deceased to bless the body,” he said.

While Milan archdiocese is limiting priests from the traditional blessing of the body in the home of the family, the funeral Mass and burial rites will be able to take place at a church or “preferably” at a cemetery, Antonelli added. 

During the nearly two months without Masses and funerals, dioceses in northern Italy have been maintaining telephone lines for grieving families with spiritual counsel and psychological services. In Milan, the service is called “Hello, is this an angel?” and is operated by priests and religious who spend time on the phone with the sick, the mourning, and the lonely. 

Aside from funerals, public Masses will still not be allowed throughout Italy under the government’s May 4 coronavirus restrictions. As Italy eases its lockdown, it remains unclear when public Masses will be allowed by the Italian government.

Italian bishops have been critical of Prime Minister Giuseppe Conte’s latest coronavirus measures, announced on April 26, saying that they “arbitrarily exclude the possibility of celebrating Mass with the people."

According to the prime minister’s April 26 announcement, the easing of lockdown measures will allow retail stores, museums, and libraries to reopen beginning May 18 and restaurants, bars, and hair salons June 1.

Movement between Italian regions, within regions, and within cities and towns is still prohibited except under strict cases of necessity.

In a letter April 23, Cardinal Gualtiero Bassetti of Perugia, the president of the Italian bishops' conference, wrote that “the time has come to resume the celebration of the Sunday Eucharist, and church funerals, baptisms and all the other sacraments, naturally following those measures necessary to guarantee security in the presence of more people in public places.”

Professional experience as a teacher and personal relationships on the field equipped one long-term worker to teach at a start-up English-speaking school.

MAY is normally a busy month behind the scenes for Gareth Chalmers and the ongoing uncertainty around sport hasn’t changed that a huge amount.

Big Zuu is headed back out on tour - but not as we know it.

There's a new focus on getting more training to early-childhood educators, but there's a dearth of information about what constitutes a high-quality, teacher-preparation program.

In this lesson, 7th grade English/language arts teacher Emily Park-Friend takes her students through a three-step interview activity.

Two high-powered N.Y. officials have put out a blueprint for overhauling teaching in the state, aiming for more-coherent policies for the profession.

The newly formed group, which plans to challenge the Council for Accreditation of Educator Preparation for market share, wants feedback from the public on its proposed standards and processes.

The current system for accrediting schools of education isn't working, argue two former senior U.S. Department of Education officials. They think school districts and philanthropists can help.

Maternal obesity is an increasingly important public health concern and may adversely affect central nervous system development in offspring. However, few studies have explored the relationship between maternal prepregnancy BMI and neurodevelopmental outcomes in children, with conflicting results.

Using data from a large and contemporary UK birth cohort, we found that maternal prepregnancy BMI is negatively associated with children’s cognitive performance. The relationship appears to become stronger as children get older, although the overall effect size is modest. (Read the full article)

The importance of transition from pediatric to adult health care for youth with special health care needs has gained increasing attention over the past decade, but fewer than half of this population received needed transition preparation in 2005–2006.

This study reports on transition findings from the 2009–2010 National Survey of Children with Special Health Care Needs and finds no discernible improvements since 2005–2006. New clinical recommendations and care processes should help to accelerate transition improvements in the future. (Read the full article)

Physical activity interventions aimed at improving body composition in childhood have had limited success and often targeted overweight children. Therefore, the efficacy of physical activity randomized controlled trials in improving body composition among children with varying adiposity levels remains unknown.

This randomized controlled trial demonstrated that a physical activity program designed to meet daily physical activity recommendations can improve cardiorespiratory fitness, decrease total fat mass, and prevent accumulation of central adiposity in a group of children with varying adiposity levels. (Read the full article)

Adolescents with chronic disease are a diverse population with common needs for transition. Disease-specific interventions have shown promise at improving patient outcomes but with substantial personnel and resource costs. Whether a generic approach across diseases may be useful is unknown.

This study is among the first to evaluate a generic (across disease) approach to transition of adolescents to adult care. The approach demonstrated promise and cost savings due to reduced personnel requirement and use of low-cost technology dissemination methods. (Read the full article)

Available bowel preparation solutions for colonoscopy continue to represent a challenge for children and their families due to poor taste, high volume, and dietary restrictions with subsequent poor compliance and need to place nasogastric tube for administration.

Low-volume polyethylene glycol (PEG) preparations and sodium picosulphate plus magnesium oxide and citric acid (NaPico+MgCit) are noninferior to PEG 4000 with simethicon for bowel preparation before colonoscopy in children. Given its higher tolerability and acceptability profile, NaPico+MgCit should be preferred in children. (Read the full article)

Preparticipation physical evaluations (PPEs) are considered necessary for a high standard of care for US scholastic athletes. However, important questions remain regarding consistency of implementation and content of cardiovascular screening practices among states.

Our results show that PPE policies are variable among US states, and adoption of current PPE-4 best practices is slow, demonstrating the need for nationwide PPE standardization. (Read the full article)

Both obesity and developmental disabilities have increased in recent decades; however, the full long-term effects of prepregnancy obesity on a child’s psychosocial development remain unknown. Limited studies suggest associations between maternal prepregnancy obesity and child psychosocial development.

This study in 6-year-old children provides evidence that severe prepregnancy obesity is associated with adverse child psychosocial outcomes, including attention-deficit/hyperactivity disorder. These findings were not explained by many pregnancy and postpartum factors related to maternal obesity or child development. (Read the full article)

When can schools use federal funds to help students with disabilities prepare for life after special education? A new resource from the federal education department offers a road map.

About two dozen teachers from across the country spent a week wrestling with questions about how to remember the Confederacy.

Teacher-preparation programs need to better prepare teachers, and especially white teachers, to serve students and communities of colors more effectively, the report says.

The National Council on Teacher Quality has found that the number of elementary programs teaching scientifically based reading instruction is increasing.

Nearly every state in the nation has experienced enrollment declines, with some states seeing steep declines of more than 50 percent.

The National Council on Teacher Quality has found that the number of elementary programs teaching scientifically based reading instruction is increasing.

Many teachers leave preservice training without clarity on what the cognitive science says about how students learn to read.

The experiences that Chris Eggerman, a 2017 graduate, had while at Penn State gave him a glimpse of the challenges and rewards he would enjoy in a career, and, paired with his education in the College of Information Sciences and Technology, equipped him with the skills he uses in his current profession as a security adviser at Liberty Mutual.

Many teachers leave preservice training without clarity on what the cognitive science says about how students learn to read.

Relaxed travel begins at home. This also includes the correct packing of your luggage.

Intel has released some benchmarks for its next-gen Core i9 'Cascade Lake-X' processors, which will be arriving next month with a big price cut. The company is also slightly dropping prices on Core S-series chips that lack GPUs, and preparing to launch the Xeon W-2200 series.

Seven universities are making major changes to how they train future principals, as part of $48.5 million Wallace Foundation initiative to redesign university-based principal-preparation programs, according to a new report from RAND.

Want to encourage girls to become professional technologists? Here's how to make sure your girls-in-tech events help them to accurately set their Science, Technology, Engineering, and Mathematics (STEM) career expectations.

The flu season has started here in the U.S., and according to the Centers for Disease Control and Prevention (CDC) data it has caused 214 deaths in the first week of 2020. Is this number higher, or lower, than usual? When does the flu season start, and how long does [...]

The post How to prepare (your graphs) for flu season appeared first on Graphically Speaking.

A new worker is able to use his profession to reach out in Central Asia.

Jio Rs. 2,399 prepaid recharge plan brings 2GB daily high-speed data benefits along with unlimited voice calling benefits for 365 days. Jio has also introduced new add-on packs at Rs. 151, Rs. 201,...

Umar Akmal was banned from all forms of cricket for three years by the PCB after pleading guilty to failing to report corrupt approaches.

Bhaat literally means rice and bhaja means fry; bhaat bhaja is the home-style fried rice among Bengalis. This recipe needs almost nothing for preparation; tastes best with leftover rice.

Democratic Party's presumptive presidential nominee and former US vice president Joe Biden alleged on Friday that President Donald Trump utterly failed to prepare for the COVID-19 pandemic and said...

With hurricane season about to begin, Delaware agricultural authorities are urging farmers and pet owners to plan ahead and prepare for weather emergencies. Hurricane Preparedness Week runs from May 26 to June 1, with hurricane season beginning June 1. The Department of Agriculture’s Delaware Animal Response Program works with state emergency officials and animal welfare organizations to assist animal owners with preparation, sheltering in place and evacuation. “Preparing and acting early can save animals’ lives,” said Secretary of Agriculture Ed Kee. “The time to take action is now, before storms create disaster conditions.”

A wildfire crew led by the Delaware Forest Service is near completion of a two-week assignment on the Fork Complex Fire, a 28,736-acre blaze near Hayfork, California in the Shasta-Trinity National Forest. Almost 2,400 personnel are battling the fire that is currently 26 percent contained. The Fork Complex is one of several large wildfires in Northern California that together cover more than 223,000 acres, one of the major factors that prompted the National Interagency Fire Center (NIFC) to increase its National Preparedness Level today to the maximum of 5 on a 5-point scale.

Livestock owners should prepare now for possible weather effects from Hurricane Joaquin, Delaware agricultural authorities said Thursday. "Being prepared is never wasted effort," said Delaware Secretary of Agriculture Ed Kee.

DOVER – The Delaware Supreme Court, by decision dated Dec. 7, 2017 in the matter of Bridgeville Rifle & Pistol Club, LTD, overturned a Superior Court ruling and ended a ban on visitors’ possessing firearms in Delaware’s state parks and state forests. Pursuant to the ruling, visitors may now possess firearms unless they are prohibited by law due to a past conviction for a violent crime, active Protection from Abuse Order, or mental health commitment. Following the Court’s decision to invalidate the existing regulations limiting firearms in state parks and forests, and some Division of Fish & Wildlife properties, the Delaware Department of Natural Resources and Environmental Control and the Delaware Department of Agriculture (DDA) anticipate issuing interim firearm regulations. The interim regulations would respect the State Constitutional provisions on firearms cited by the Court, by implementing the State’s interest in ensuring the safety of the public in a less restrictive manner.

About 1 in 6 Americans gets sick every year from foodborne illnesses. Under the Food Safety Modernization Act (FSMA), FDA and the USDA work closely to trace the cause and origin of these illnesses. Traceability is key so the origin is quickly detected, and a recall enacted, preventing others from becoming ill. Preparation and planning are necessary for produce growers and businesses in the supply chain to successfully navigate a product recall. The Delaware Department of Agriculture’s Produce Safety Program will present the United Fresh Recall Ready Workshop on January 22, 2020 at Dover Downs from 8 a.m. to 5 p.m.

As a long-time SAS 9 programmer, I typically accomplish my data preparation tasks through some combination of the DATA Step, Proc SQL, Proc Transpose and some housekeeping procs like Proc Contents and Proc Datasets. With the introduction of SAS Viya, SAS released a new scripting language called CASL – a [...]

Challenge accepted: Learning data prep in CASL was published on SAS Users.