Differential detergent fractionation of cells is a rapid method for extraction of cytoplasmic and nuclear proteins in preparation of an immunoprecipitation. This method can be applied for use of adherent or suspension cells and can significantly reduce nonspecific background in an immunoprecipitation by separation of cellular compartments into individual fractions. The lysis of cells by differential detergents permits the rapid extraction of proteins from the cytoplasm (digitonin), the cytoplasmic membranes, and organelles (Triton X-100), and nucleoplasm (Tween/DOC), facilitated through the use of distinct extraction buffers. Cytoplasmic and nuclear matrix proteins as well as DNA are left behind during the detergent-based extraction.

Bacterial artificial chromosome (BAC) clones are rendered electrocompetent and transformed with the recombinant shuttle vector, pLD53SCAB/AB-box. Cointegrates are selected by growth on chloramphenicol and ampicillin to ensure recombination of the shuttle vector into the BAC.

A protein shredder that occurs in cell membranes of brain cells apparently also indirectly regulates the fat metabolism.

Acute myeloid leukemia is an aggressive cancer of the blood-forming system. It affects the hematopoietic stem cells, or blood stem cells, of various white blood cells and of the red blood cells and platelets.

With a discovery that could rewrite the immunology textbooks, an international group of scientists, including the teams of Bart Lambrecht, Martin Guilliams, Hamida Hammad, and Charlotte Scott (all from the VIB-UGent Center for Inflammation Research) identified a new type of antigen-presenting immune cell.

Vowing to return next summer, this year's Little League World Series was cancelled due to the coronavirus pandemic. All seven World Series tournaments and their respective regional qualifying events are off this summer.

Winnipeg will soon test "small cell" technology to improve cellular reception in parts of Winnipeg, ahead of a possible future transition to 5G service. 

Quebec Health Minister Danielle McCann says the government is evaluating whether to allow the world figure skating championships to go ahead next week in Montreal.

The world figure skating championships, scheduled for March 18-22 in Montreal, have been cancelled because of the spread of COVID-19.

With this year's worlds cancelled because of the COVID-19 pandemic, Rob Pizzo looks at the last time the event didn't happen.

Hope for a 2020 Grey Cup in Regina is slowly dwindling as the CFL hints at a season cancellation. 

The world women's curling championship in Prince George, B.C. has been cancelled because of the outbreak of COVID-19.

The men's curling world championship in Glasgow was cancelled Saturday morning to prevent the spread of COVID-19. Brad Gushue and his rink from Newfoundland and Labrador was going to represent Canada after winning the Brier.

When the World Curling Federation cancelled the women's, men's and mixed doubles championships set for March and April, it said it would be discussing potential options including rescheduling the events — perhaps playing them in the fall at one venue. CBC Sports has learned that will no longer be the case.

Purifying and culturing cells from the central nervous system (CNS) has proved to be an incredibly powerful tool for dissecting fundamental neuron and glial properties, and especially powerful in understanding neuronal–glial interactions. In a series of detailed protocols, we have provided step-by-step instructions for purifying and culturing specific types of neurons, glia, and vascular cells from the CNS by immunopanning. This article discusses common pitfalls and errors as well as important design considerations for the immunopanning procedure.

COMPLAINTS about holidays have overtaken job concerns as the number one issue for hundreds of Scots contacting two independent national advice services every day during the coronavirus crisis.

The latest iPad Pro featuring 1TB of storage and cellular connectivity is on sale at Amazon for just $1,299.99. Models with less storage and just Wi-Fi are also marked down.

THE Open Championship will not take place at all this year.

Glasgow's Summer Nights at the Bandstand festival has become the latest major music event to be cancelled due to the Covid-19 pandemic.

THIS year, more than ever, we need to see a love story played out on stage. We need to see Covid-19 killed off as convincingly as Snow White’s wicked step ma. We need lines such as: “He’s two metres behind you!” Or the Uglies throwing scorchers at each other like: “Did you get that face mask in Poundstretchers?” “Whit? A’m no’ wearing a face mask, ya cheeky madam.”

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.

A number of studies have examined the early impact of the varicella vaccination program on varicella-related hospitalizations and have found evidence of decline after vaccine implementation.

This study further documents the continued decline in varicella-related hospitalizations during the 1-dose varicella vaccination era and demonstrates statistically significant declines of >65% in all age groups. These data suggest that varicella vaccination prevented ~50 000 hospitalizations from 2000 to 2006. (Read the full article)

Children with sickle cell disease are at high risk of complications from influenza infection and have been recommended to receive annual influenza vaccine since the 1970s. Few safety studies, however, have examined the safety of influenza vaccine in this population.

This large cohort study did not find an association between influenza vaccination and hospitalization for sickle cell crises in children with sickle cell anemia. (Read the full article)

Children undergoing stem cell transplantation are thought to be at risk for increased distress, adjustment difficulties, and impaired health-related quality of life. Few interventions to improve adjustment and quality-of-life outcomes in this setting have been tested.

The excellent outcomes observed in all patient groups, including controls, may be a result of improvements in standard supportive care. Stem cell transplantation may not be as demanding as previously thought to be, and children undergoing this procedure appear resilient to the challenge. (Read the full article)

Sending and receiving sexually explicit picture and text messages via cell phone (ie, "sexting") among adolescents is publicized as a societal and public health concern, yet it is unknown whether sexting is associated with physical sexual activity or sexual risk behavior.

This study is the first to examine sexting among a probability sample of adolescents and found that sexting is associated with sexual activity, sexual risk behavior, and knowing other person(s) who have sent a sext. (Read the full article)

Children with sickle cell anemia are at risk of central nervous system damage, including stroke. Even children without evidence of abnormality on neuroimaging are at risk of significant declines in neurocognitive function, starting at early ages.

This study adds the observation that poorer neurocognitive and behavioral function is associated with older age in infants and toddlers with sickle cell anemia, much earlier than previously expected. (Read the full article)

Children with sickle cell disease (SCD) have a higher basal metabolic rate, and have historically been underweight. In the general pediatric population, the average BMI percentile has been rising over the past 2 decades.

BMI percentiles for children with SCD in New England are higher than historically reported, mimicking the weight status in the general pediatric population. In children with SCD, higher hemoglobin levels increased the odds of being overweight and obese. (Read the full article)

Varicella vaccine is known to be highly effective, with added benefit from a second dose.

This study demonstrates the lasting effectiveness of varicella vaccine and the benefit of the second dose. Breakthrough varicella occurred soon after vaccination, varicella rates did not increase over 14 years, and there was no increase in zoster in the cohort. (Read the full article)

Before the introduction of conjugate pneumococcal vaccines and routine penicillin prophylaxis, febrile patients with sickle cell disease were known to have a 3% to 5% risk of bacteremia. Consequently, hospitalization rates for febrile episodes are >70%.

We observed no mortality or morbidity among those managed completely as outpatients, and bacteremia occurred in <1%. Physicians should strongly consider outpatient management of febrile children with sickle cell disease if there are no other indications for admission. (Read the full article)

The United States switched from whole-cell to acellular pertussis vaccines during the 1990s. Whether pertussis risk during a California outbreak differed between teenagers who previously received whole-cell or acellular pertussis vaccines early in life has not been reported.

We evaluated pertussis risk in 10 to 17 year olds at Kaiser Permanente Northern California during a recent pertussis outbreak. Those given whole-cell pertussis vaccines in childhood were more protected than those given acellular pertussis vaccines. (Read the full article)

Persons with sickle cell anemia are known to have increased medical expenses, but little is known about the effects of hydroxyurea treatment on costs. In adults with severe sickle cell anemia, hydroxyurea has been reported to reduce expenses from hospitalization.

In this randomized placebo-controlled prospective multicenter trial of hydroxyurea in very young children with sickle cell anemia, not selected for severity, hydroxyurea was associated with significant medical cost savings due to a reduction in hospitalization expenses. (Read the full article)

The 1-dose childhood varicella vaccination program in the United States resulted in dramatic declines in varicella incidence, hospitalizations, and deaths. There is little information on the impact of the 2006 recommendation for 2-dose varicella vaccination of children on varicella epidemiology.

In the first 5 years of the 2-dose varicella vaccination program, declines in varicella incidence were seen in all age groups, including infants who are not eligible for varicella vaccination, providing evidence of the benefit of high population immunity. (Read the full article)

As more children with sickle cell disease survive into adulthood, they are increasingly hospitalized in both children’s and general hospitals and managed by different provider specialists. But it is unknown if hospital type and provider specialty affect patient outcomes.

Using a large national administrative dataset, this study revealed that general hospitals were associated with higher rates of intubation and longer lengths of stay compared with children’s hospitals for adolescents and young adults with SCD admitted with acute chest syndrome. (Read the full article)

Vaso-occlusive episodes (VOEs) are a common complication of sickle cell disease, resulting in morbidity. Magnesium is a vasodilator and has been shown to improve red blood cell hydration. Previous small studies have suggested that treatment with magnesium may decrease VOEs.

Intravenous magnesium sulfate is well tolerated in relatively high doses but had no effect on the length of stay in hospital, pain scores, or cumulative analgesia used in children admitted with painful VOEs in sickle cell disease. (Read the full article)

Currently, the false-positive rate of appendicitis in children is ≤5%. Abdominal imaging and blood tests (particularly leukocytosis) help minimize the negative appendectomy rate, but appendicitis is not always associated with an elevated white blood cell count.

Reducing the threshold of leukocytosis as a criterion for appendicitis to 8000 to 9000 white blood cells per µL improves specificity (negative appendectomy: <1%) while only marginally decreasing sensitivity. (Read the full article)

Waning effectiveness of 5 doses of acellular pertussis vaccines is well documented after 6 years of age, but data are lacking for fewer doses in younger children.

In 2- to 3-month-old infants, 1 dose of the diphtheria–tetanus–acellular pertussis vaccine gave significant protection against hospitalized pertussis. The effectiveness of 3 doses decreased from 84% between 6 and 11 months to 59% after 3 years. (Read the full article)

The American Academy of Pediatrics has recommended that sibling donors should have an independent advocate. Defining the need for and role of this advocate is hampered by a lack of empirical data.

This study provides prospective family data regarding siblings’ experiences during HLA typing and donation pre- and posttransplantation. Most family members, including the siblings, perceive no choice in typing or donation, yet have few concerns and report positive aspects to participating. (Read the full article)

Varicella vaccine is effective, but there is concern that widespread use in young children may lead to a shift in the age of infection, with potentially more severe disease later in childhood and adolescence.

This study documents that varicella vaccine resulted in a decline of varicella incidence and hospitalization in all age groups, with no shift to older age groups. (Read the full article)

Indoor tanning has gained widespread popularity among adolescents and young adults. Incidence rates of early-onset basal cell carcinoma also appear to be rising. Scant evidence exists on the impacts of early exposure and whether it leads to early occurrence of this malignancy.

In a US population-based study, indoor tanning was associated with an elevated risk of basal cell carcinomas occurring at or before the age of 50 years, with an increasing trend in risk with younger age at exposure among adolescents and young adults. (Read the full article)

Obstructive sleep apnea syndrome (OSAS) prevalence in children with sickle cell anemia is not well described. Although these children often experience nocturnal oxygen desaturation, it is unclear whether they are more likely to have OSAS.

Children with sickle cell anemia have a high prevalence of OSAS with typical symptoms, beyond just nocturnal oxyhemoglobin desaturation. This study supports the need for increased efforts to screen for, diagnose, and treat OSAS in this vulnerable population. (Read the full article)

Growth impairment in sickle disease has been a consistent finding in published reports. Hydroxyurea (HU) decreases vasoocclusive events and increases hemoglobin levels, which may improve growth. However, HU may adversely affect growth in young children by its effect on DNA synthesis.

Height, weight, and head circumference were normal in HU-treated children in the study as compared with the World Health Organization standards. Height, weight, and BMI z scores were similar in placebo and treatment groups. There were no harmful effects of HU on growth. (Read the full article)

Hydroxyurea is a treatment option for young patients with sickle cell disease (SCD). Establishing the safety of hydroxyurea is of paramount importance. The effect of hydroxyurea on immune function and immunizations in SCD has not been studied previously.

Children with SCD receiving hydroxyurea have lower lymphocyte, CD4, and memory T-cell counts compared with those receiving placebo, but still in the range for healthy children. Despite slower response to measles vaccine, measles, mumps, and rubella and pneumococcal vaccines are effective. (Read the full article)

Germ cell tumors in children are heterogeneous and rare neoplasms that occur in various locations, such as gonads, the central nervous system, and the pelvis. The incidence rate has been increasing in some countries.

Population-based analyses of germ cell tumors in children are rare. This population-based study describes the incidence rates, trends, and survival of germ cell tumors in German children from 1987 to 2011. (Read the full article)

Patients with sickle cell disease frequently express dissatisfaction with emergency department treatment of painful crises. Time to opioid administration has been suggested as a quality of care measure for painful crises.

Although not associated with hospital admission, time to opioid administration in sickle cell disease painful crises was associated with secondary outcomes including improvement between the first 2 pain scores, decreased pain score area under the curve at 4 hours, decreased emergency department length of stay, and increased total opioids. (Read the full article)

This is the first population-based study regarding the epidemiologic characteristics of pediatric zoster among only those who had contracted varicella.

The herpes zoster (HZ) incidence among only children with varicella infection is higher than previously reported. The HZ incidence increased for children contracting varicella aged <2 years. After a vaccination program, the HZ risk increased for those contracting varicella aged ≥2 years. (Read the full article)

Tous les éléments d’une confrontation violente sont en place en Burundi. En observant les derniers développements, il semble que les éléments qui ont conduit par le passé à des massacres et à une longue guerre civile au début des années 1990 se remettent en place.

Antimicrobial peptides and proteins play critical roles in the host defense against invading pathogens. We recently discovered that recombinantly expressed human and mouse serum amyloid A1 (rhSAA1 and rmSAA1) proteins have potent antifungal activities against the major human fungal pathogen Candida albicans. At high concentrations, rhSAA1 disrupts C. albicans membrane integrity and induces rapid fungal cell death. In the current study, we find that rhSAA1 promotes cell aggregation and targets the C. albicans cell wall adhesin Als3. Inactivation of ALS3 in C. albicans leads to a striking decrease in cell aggregation and cell death upon rhSAA1 treatment, suggesting that Als3 plays a critical role in SAA1 sensing. We further demonstrate that deletion of the transcriptional regulators controlling the expression of ALS3, such as AHR1, BCR1, and EFG1 in C. albicans results in similar effects to that of the als3/als3 mutant upon rhSAA1 treatment. Global gene expression profiling indicates that rhSAA1 has a discernible impact on the expression of cell wall- and metabolism-related genes, suggesting that rhSAA1 treatment could lead to a nutrient starvation effect on C. albicans cells.

Periodontitis as a biofilm-associated inflammatory disease is highly prevalent worldwide. It severely affects oral health and yet closely links to systemic diseases like diabetes and cardiovascular disease. Porphyromonas gingivalis as a ‘keystone' periodontopathogen drives the shift of microbe-host symbiosis to dysbiosis, and critically contributes to the pathogenesis of periodontitis. Persisters are a tiny subset of biofilm-associated microbes highly tolerant to lethal treatment of antimicrobials, and notably metronidazole-tolerant P. gingivalis persisters have recently been identified by our group. This study further explored the interactive profiles of metronidazole-treated P. gingivalis persisters (M-PgPs) with human gingival epithelial cells (HGECs). P. gingivalis cells (ATCC 33277) at stationary phase were treated with lethal dosage of metronidazole (100 μg/ml, 6 hours) for generating M-PgPs. The interaction of M-PgPs with HGECs was assessed by microscopy, flow cytometry, cytokine profiling and qPCR. We demonstrated that the overall morphology and ultra-cellular structure of M-PgPs remained unchanged. Importantly, M-PgPs maintained the capabilities to adhere to and invade into HGECs. Moreover, M-PgPs significantly suppressed pro-inflammatory cytokine expression in HGECs at a comparable level with the untreated P. gingivalis cells, through the thermo-sensitive components. The present study reveals that P. gingivalis persisters induced by lethal treatment of antibiotics could maintain their capabilities to adhere to and invade into human gingival epithelial cells, and perturb the innate host responses. Novel strategies and approaches need to be developed for tackling P. gingivalis and favourably modulating the dysregulated immuno-inflammatory responses for oral/periodontal health and general wellbeing.