Ste50 adaptor protein influences Ras/cAMP-driven stress-response and cell survival in Saccharomyces cerevisiae.
The Ste50 adaptor protein is involved in a variety of cellular pathways that yeast cells use to adapt rapidly to environmental changes. A highly activated Ras-cyclic AMP (cAMP) pathway by deletion of the high-affinity cAMP-dependent phosphodiesterase 2 (PDE2) leads to repression of a stress mediated response and cell survival. Here we show that inactivation of STE50 confers a synthetic genetic interaction with pde2Delta. A hyperosmotic stress growth defect of ste50Delta pde2Delta cells is exacerbated by extracellular cAMP or by galactose as the sole carbon source in the medium. The inactivation of the serine/threonine protein-kinase Akt homologue Sch9 increase stress resistance and extend chronological life span. By pde2Delta-dependent increase of the Ras-cAMP pathway activity, inactivation of STE50 results in an extreme shortening of life span and oxidative stress sensitivity of sch9Delta mutants. Furthermore, sch9Delta can promote transcription of the small heat shock protein HSP26 in a PDE2-dependent manner; however, sch9Delta can promote transcription of the mitochondrial superoxide dismutase SOD2 in a PDE2- and STE50-dependent manner. These data indicate that inactivation of STE50 influences stress tolerance in mutants of the Ras-cAMP pathway, which is a major determinant of intrinsic stress tolerance and cell survival of the Saccharomyces cerevisiae.


Effectiveness of intervention to implement tobacco cessation counseling in community chain pharmacies.
To test the feasibility of implementing ask-advise-refer (AAR) tobacco cessation counseling approach in community chain pharmacies serving low socioeconomic areas and to assess the effectiveness of a multimodal intervention on short-term implementation of AAR. Randomized controlled trial. South-central Wisconsin from July 2008 through March 2009. Pharmacists and technicians from 16 community chain pharmacies. Training to implement AAR, workflow integration recommendations, tobacco cessation poster to create awareness, and a support visit. Number of pharmacy patrons asked about tobacco use, number of tobacco users advised to quit, number of tobacco users enrolled in the quit line, and number of quit line cards given. As hypothesized, the multimodal intervention significantly predicted the number of patrons asked to quit (estimate 4.84, incidence rate ratio 127.2, P < 0.001), number of tobacco users advised to quit (2.12, 8.33, P < 0.01), number of tobacco users enrolled in the quit line (2.31, 10.13, P < 0.001), and number of quit line cards given (1.04, 2.82, P < 0.05). This trial demonstrates the feasibility of implementing AAR in routine community pharmacy practice. This trial also supports the short-term effectiveness of the multimodal intervention in facilitating AAR in partnership with other public health systems. More research is needed to evaluate the generalizability, effectiveness, and sustainability of AAR, including factors influencing adoption and the impact on cessation.


Neural anatomy of the radial forearm flap.
Typically the lateral antebrachial cutaneous nerve alone is used to innervate the radial forearm free flap when a sensate flap is required. The authors desired, by means of fresh cadaveric microdissections and by means of local anesthetic injections in living subjects, to map the sensory nerve territories of this flap. Eight radial forearm flaps were elevated and the medial antebrachial cutaneous nerve (MABC), lateral antebrachial cutaneous nerve (LABC), and superficial radial sensory nerve (SRSN) were dissected with the aid of an operating microscope (2.5-10x) and traced to their dermal insertions. In the injection study, the MABC, LABC, and SRSN in eight forearms of 4 subjects were blocked sequentially with 2% lidocaine injections. The resulting sensory deficit from each injection was mapped on the skin and superimposed on the marked radial forearm flap territory. Distribution of the three dissected nerve regions and the sensory deficit after injection were determined by digital images and computer analysis. During flap dissections, mean nerve distributions of total flap area were as follows: LABC, 61.8% (range, 48.3-71.6%); MABC, 33.8% (range, 30.5-38.9%); and SRSN, 34.6% (range, 26.8-44.1%). After nerve block the mapped sensory areas were as follows: LABC, 62.3% (range, 44.5-88.5%); MABC, 19.6% (range, 8.0-35.8%); and SRSN, 19.5% (range, 9.9-26.3%). At least 40% of the total flap area was not innervated by the LABC as identified both by nerve dissection and sensory local anesthetic blockade. By including the LABC, MABC, and SRSN in the radial forearm flap, both the theoretical and the clinically determined useful sensory innervation of the radial forearm flap potentially would be increased.


Precice Stryde® Magnetic Internal Lengthening Nail does not Impair Bone Healing Despite Radiographic and Clinical Symptoms.
The Precice Stryde® internal magnetic lengthening nail allowed many patients a full weight-bearing experience during femur and tibia lengthening, but concerns over corrosion, pain and radiographic changes led to the implant's recall. Despite the recall, it is important to understand the rate of these occurrences and their influence on the overall success of the lengthening procedure. We aimed to investigate radiographic changes, patient-reported symptoms and bone healing indices for our cohort of Stryde lengthening. Our surgical database and electronic medical record system were used to review and document patient demographics, indications for lengthening, length achieved, bone healing index (BHI), location and type of radiographic changes, time until radiographic changes were first visible, presence of pain symptoms (not attributable to surgery or distraction), time to implant removal and if the pain symptoms resolved following implant extraction. From January 2019 to February 2021, 90 Stryde nails (78 femur and 12 tibia) were implanted in 63 patients. The cohort included 48 males and 15 females. The average length [± standard deviation (SD)] achieved was 58.4 ± 22.7 mm. The 66 bones (73%) developed radiographic changes and were found to be 58/78 (74%) femurs and 8/12 (67%) tibias. The average time to initial radiographic changes was 168 ± 108.1 days (femur) and 276 ± 126.8 days (tibia). Late-onset pain developed in 10 femur lengthening (11.1% of all nails) surgeries across eight patients (12.7% of all patients). All patients' pain resolved; three instances prior to nail removal and the remaining seven after nail removal. No patients were re-presented with worsening pain or radiographic changes following implant removal. Radiographic or symptomatic abnormalities did not impair bone formation. The BHI for femurs with (29.6 ± 16.6 days/cm, n = 58) vs without (29.4 ± 17.9 days/cm, n = 20) radiographic or symptomatic irregularity were nearly identical (p = 0.961). The difference between BHI for tibias with (39.3 ± 7.8 days/cm, n = 8) vs without (86.1 ± 38.2 days/cm, n = 4) radiographic changes was influenced by outliers and underpowered to draw a conclusion. Bone lengthening with the Stryde nail was associated with high rates of radiographic abnormalities, but symptoms were uncommon and resolved with explantation. The radiographic changes did not affect bone healing in the femur. Radiographic changes including bone hypertrophy and osteolysis were common after bone lengthening with the Stryde nail, but the development of pain following consolidation was rare and resolved with implant removal.The BHI in femurs was not affected by radiographic changes. Reif TJ, Geffner A, Hoellwarth JS, et al. Precice Stryde® Magnetic Internal Lengthening Nail does not Impair Bone Healing Despite Radiographic and Clinical Symptoms. Strategies Trauma Limb Reconstr 2023;18(2):94-99.


An efficient parallelization of phosphorylated peptide and protein identification.
Protein sequence database search based on tandem mass spectrometry is an essential method for protein identification. As the computational demand increases, parallel computing has become an important technique for accelerating proteomics data analysis. In this paper, we discuss several factors which could affect the runtime of the pFind search engine and build an estimation model. Based on this model, effective on-line and off-line scheduling methods were developed. An experiment on the public dataset from PhosphoPep consisting of 100 RAW files of phosphopeptides shows that the speedup on 100 processors is 83.7. The parallel version can complete the identification task within 9 min, while a stand-alone process on a single PC takes more than 10 h. On another larger dataset consisting of 1,366,471 spectra, the speedup on 320 processors is 258.9 and the efficiency is 80.9%. Our approach can be applied to other similar search engines.


Prevalence and Risk Factors of Diabetes Mellitus in Patients with Autoimmune Hepatitis.
The administration of corticosteroids is a standard treatment for autoimmune hepatitis (AIH), but it can occasionally induce various adverse effects. Diabetes mellitus (DM) is a major complication of chronic liver diseases. We investigated the prevalence and risk factors of DM in patients with AIH. We retrospectively analyzed 118 Japanese patients diagnosed with AIH from 1990 to 2014 at our institution. The prognosis of patients with and without DM was also compared. Twenty-nine (24.5%) patients had DM and 21 (72.4%) received corticosteroids. The annual cumulative incidence rate of newly diagnosed DM was 1.2%. Multivariate analysis showed that DM occurred in older patients [OR=6.290; 95% confidence interval (CI)=1.230-32.100; p=0.018] with higher serum immunoglobulin G levels (OR=12.400; 95% CI=2.560-60.400; p=0.002). A Cox hazard regression analysis revealed that predictive factors for DM were absence of other autoimmune diseases (OR=0.171; 95% CI=0.036-0.805; p=0.025), use of corticosteroids (OR=6.693; 95% CI=1.391-32.210; p=0.049) and lower platelet counts (OR=3.873; 95% CI=1.021-14.690; p=0.046). The 10 year survival rates of the DM and non-DM groups were 94.1% and 94.6%, respectively. There was no significant difference between these groups (p=0.293). DM occurred in 24.5% of patients with AIH; older age, absence of other autoimmune diseases and higher serum immunoglobulin G levels are risk factors. Taking corticosteroids and a lower platelet count are risk factors for a new onset of DM. DM did not influence the prognosis of AIH patients.


Feeding of prohibited substances (swill) to pigs in Australia.
To assess current swill feeding legislation, swill feeding investigation practices by authorities and feeding practices of pig producers who trade via saleyards in eastern Australia in order to determine levels of understanding and conformance related to current swill feeding legislation. A three-tiered approach was undertaken to gather information on the feeding of prohibited substances (swill) to pigs in Australia. Firstly, a review of swill feeding legislation was undertaken to highlight the commonalities and inconsistencies between the various state and territory legislations in defining swill. Secondly, agricultural authorities were contacted in each state to gather information on swill feeding investigations undertaken in 2006. Finally, face-to-face interviews were conducted with 106 pig producers who traded pigs at one of six saleyards in eastern Australia to ascertain their knowledge of swill feeding and to determine the feeding practices of this sector of the industry. Areas of concern identified included (1) inconsistencies in the feedstuffs classed as 'swill' among states, (2) the number of producers who had been prosecuted for swill feeding in 2006 (n = 4 of 148 inspections), (3) the low knowledge base of producers who sell pigs at saleyards regarding swill feeding, and (4) the types of feedstuffs provided to pigs marketed at saleyards. Our findings highlight the need for a consistent definition for 'swill' across Australian states and for improved awareness of swill feeding among producers, particularly those who market pigs at saleyards.


Piloting a new approach to the treatment of obesity using dexamphetamine.
There is a clear need for a new approach to the treatment of obesity, which is inexpensive and is effective for establishing lifestyle change. We conducted a pilot study to evaluate whether dexamphetamine can be used safely, combined with diet and exercise, for treating obesity. Our ultimate aim is to develop a 6-month treatment program for establishing the lifestyle changes necessary for weight control, utilizing dexamphetamine for its psychotropic effect on motivation. We viewed the anorexigenic effect as an additional advantage for promoting initial weight loss. Obese adults were treated with dexamphetamine for 6 months (maximum of 30 mg twice daily), diet, and exercise. Weight, electrocardiogram, echocardiogram, and blood pressure were monitored. Twelve out of 14 completed 6 months treatment. Weight loss by intention to treat was 10.6 kg (95% CI 5.8-15.5, p < 0.001). The mean weight gain in the 6 months after ceasing dexamphetamine was 4.5 kg (95% CI 1.9-7.2, p = 0.003), leaving a mean weight loss at 12 months from baseline of 7.0 kg (95% CI -13.4 to -0.6, p = 0.03). All reported favorable increases in energy and alertness. Dose-limiting symptoms were mood changes (2) and insomnia (2). None had drug craving on ceasing dexamphetamine, and there were no cardiac complications. Among the seven women, there was a significant correlation for those who lost most weight on treatment to have the least regain in the following 6 months (r = 0.88, p = 0.009). Our treatment with dexamphetamine, diet, and exercise was well tolerated and effective for initial weight loss. Future research will focus on identifying baseline predictive variables associated with long-term weight control.


Transcriptional changes associated with lack of lipid synthesis in parasitoids.
Phenotypic regression of morphological, behavioral, or physiological traits can evolve when reduced trait expression has neutral or beneficial effects on overall performance. Studies on the evolution of phenotypic degradation in animals have concentrated mostly on the evaluation of resulting phenotypes, whereas much less research has been dedicated to uncovering the molecular mechanisms that underlie phenotypic regression. The majority of parasitoids (i.e., insects that develop on or inside other arthropods), do not accumulate lipid reserves during their free-living adult life-stage and represent an excellent system to study phenotypic regression in animals. Here, we study transcriptional patterns associated with lack of lipogenesis in the parasitic wasp Nasonia vitripennis. We first confirmed that N. vitripennis does not synthesize lipids by showing a reduction in lipid reserves despite ingestion of dietary sugar, and a lack of incorporation of isotopic labels into lipid reserves when fed deuterated sugar solution. Second, we investigated transcriptional responses of 28 genes involved in lipid and sugar metabolism in short- and long-term sugar-fed females relative to starved females of N. vitripennis. Sugar feeding did not induce transcription of fatty acid synthase (fas) or other key genes involved in the lipid biosynthesis pathway. Furthermore, several genes involved in carbohydrate metabolism had a lower transcription in fed than in starved females. Our results reveal that N. vitripennis gene transcription in response to dietary sugar deviates markedly from patterns typically observed in other organisms. This study is the first to identify differential gene transcription associated with lack of lipogenesis in parasitoids and provides new insights into the molecular mechanism that underlies phenotypic regression of this trait.


Reading skills in children diagnosed with hyperlexia: case reports.
Hyperlexia is characterized by spontaneous and early acquisition of reading skills, manifested before the age of five, without any formal education. Expressive and receptive language deficit, excellent memory, delayed language skills, echolalia, perseverations, and difficulty to understand verbal contexts, are common symptoms in individuals with hyperlexia and global developmental disorders, including Asperger's syndrome. The aim of this study was to describe the reading skills of individuals with hyperlexia. The participants were six boys with a history of hyperlexia, perceived by relatives before 36 months of life, chronologically aged between four years and four months to five years and two months. The following evaluation procedures were applied: interview with the parents or responsible guardians, communicative behavior observation, Peabody Picture Vocabulary Test (PPVT), recognition of alphabet letters and numbers, School Performance Test, text reading, and reading comprehension. The participants showed different performances in the PPVT, and recognized letters, numbers and isolated words. There was difficulty in answering the writing subtest; the majority did not understand the text that they read and all of them demonstrated recognition of letters and numbers. Some participants were able to read words and texts. However, all participants presented alterations in other areas of language development, such as receptive vocabulary, communicative behavior and understanding of the material read, in addition to altered behaviors of social adaptation, both interactive and restrictive.


Development of a single, practical measure of surgical site infection (SSI) for patient report or observer completion.
Surgical site infections (SSIs) are the third most common hospital-associated infection and can lead to significant patient morbidity and healthcare costs. Identification of SSIs is key to surveillance and research but reliable assessment is challenging, particularly after hospital discharge when most SSIs present. Existing SSI measurement tools have limitations and their suitability for post-discharge surveillance is uncertain. This study aimed to develop a single measure to identify SSI after hospital discharge, suitable for patient or observer completion. A three-phase mixed methods study was undertaken: Phase 1, an analysis of existing tools and semi-structured interviews with patients and professionals to establish the content of the measure; Phase 2, development of questionnaire items suitable for patients and professionals; Phase 3, pre-testing the single measure to assess acceptability and understanding to both stakeholder groups. Interviews and pre-testing took place over 12 months in 2014-2015 with patients and professionals from five specialties recruited from two UK hospital Trusts. Analyses of existing tools and interviews identified 19 important domains for assessing SSIs. Domains were developed into provisional questionnaire items. Pre-testing and iterative revision resulted in a final version with 16 items that were understood and easily completed by patients and observers (healthcare professionals). A single patient and observer measure for post-discharge SSI assessment has been developed. Further testing of the validity, reliability and accuracy of the measure is underway.


A rare cause of massive ascites: familial Mediterranean fever.
Familial Mediterranean fever is an autosomal recessive disease characterized by recurrent fever and peritoneal and pleural inflammation. It is an inherited disorder commonly found in Armenians, Turks, Arabs, Balkans, and Jews originating from North African countries. A small amount of peritoneal fluid collection can be observed during peritoneal attacks in patients with Familial Mediterranean fever, but chronic ascites has been described rarely in these patients. A 42-year-old female patient was admitted to our clinic in June 2010 with fever, severe abdominal pain and abdominal distention that had continued for one month. There was no family history of periodic fevers or abdominal pain. We could not find any cause for ascites, including tuberculosis. A diagnosis of Familial Mediterranean fever was suspected based on the clinical findings and her family history. She was screened for mutations causing Familial Mediterranean fever, and when found to be homozygous for M694V, treatment with colchicine was initiated. After treatment, the amount of ascites decreased, and relief of symptoms was confirmed during a follow-up. In conclusion, because Familial Mediterranean fever is common in our country, it should be considered in the differential diagnosis of patients with ascites of unknown etiology in populations where hereditary inflammatory disease is endemic.


Hospital standardised mortality ratio: A reliable indicator of quality of care?
This study investigates (1) whether the hospital standardised mortality ratio (HSMR) model underestimates or overestimates disease severity and (2) the completeness of the data collected by administrators to calculate HSMR in a cohort of deceased patients with the diagnosis of pneumonia. In this retrospective cohort study Pneumonia Severity Index (PSI) and Abbreviated Mortality in Emergency Department Sepsis (abbMEDS) scores and associated mortality probabilities were obtained from 32 deceased pneumonia patients over the year 2014 in the VU University Medical Centre. These were compared with mortality probabilities of the Central Bureau for Statistics (CBS) calculated for every patient using the HSMR model. Clinical charts were examined to extract relevant comorbidities to determine the reliability of data sent to the national registration of hospital care. Risk categories determined by using the PSI and the abbMEDS were significantly higher compared with the risk categories according to HSMR (p = 0.001, respectively p = 0.000). The mean difference between the number of comorbidities in our registration and the coders' registration was 1.97 (p = 0.00). The mean difference was 0.97 (p = 0.000) for the number of comorbidities of influence on the Charlson Comorbidity Index (CCI) and 1.25 (p = 0.001) for the calculated CCI. The results of this study suggest that the mortality probabilities as calculated by the CBS are an underestimation of the risk of dying for each patient. Our study also showed that the registration of data sent to the CBS underestimated the actual comorbidities of the patient, and could possibly influence the HSMR.


Infectious keratitis after astigmatic keratotomy in penetrating keratoplasty: review of three cases.
To present 3 cases in 2 patients of infectious keratitis of the astigmatic keratotomy (AK) site in patients who had previous penetrating keratoplasty. History, clinical examination, chart review, cultures, and laboratory results were obtained from 2 patients with infectious keratitis within the donor stroma of their graft after an AK was performed. One patient had an early infiltrate of the AK site that was culture negative. This cleared with fortified antibiotics, and the patient had a final visual acuity of 20/30-2. The second patient developed 2 late infections, the first with coagulase negative Staphylococcus and the second infection with Pseudomonas aeruginosa. Both of these infections cleared with fortified antibiotics, and the final visual acuity was 20/25. Neither patient developed a rejection episode from the infections, and both grafts survived. We report a series of 3 cases of infectious keratitis in 2 patients after AKs in corneal grafts. This, to the best of our knowledge, has not been previously reported. Patients who suffer infectious keratitis after AK in corneal grafts may have good visual outcomes if managed with appropriate antibiotic regimens and closely followed.


Varicella pneumonia. A report of 20 cases, with postmortem examination in six.
Twenty cases of primary varicella pneumonia, 16 in adults and four in children, were studied. Two adults and four children died, two of the latter with complicating bacterial infections. In two patients the primary cause of death was severe alveolar-capillary block. Staphylococcal septicemia, midbrain hemorrhage and meningoencephalitis were primarily responsible for death in other patients.Radiologically, the lungs showed diffuse, poorly marginated, nodular lesions, often peribronchial in location, more easily defined in the thinner peripheral lung fields, with an alveolar acinar pattern, tending to coalesce in the hilar and perihilar regions. Pathologically, the cutaneous varicella lesions were matched by similar lesions regularly found in the lungs and pleura, as well as the peritoneum and the liver. The pulmonary nodular lesions corresponded to alveoli, filled with precipitated protein and active inflammatory cellular material, surrounding the bronchioles, which themselves were often involved, and these in turn were surrounded by areas of normally aerated alveoli. Eleven moderately to severely ill patients were treated with antibiotics, six moderately to extremely ill were treated with antibiotics and adrenal cortical steroids. There was no evidence of significant change in the course of the disease resulting from use of steroid therapy.


Apolipoprotein E mRNA expression in mononuclear cells from normolipidemic and hypercholesterolemic individuals treated with atorvastatin.
Apolipoprotein E (apoE) is a key component of the lipid metabolism. Polymorphisms at the apoE gene (APOE) have been associated with cardiovascular disease, lipid levels and lipid-lowering response to statins. We evaluated the effects on APOE expression of hypercholesterolemia, APOE ε2/ε3/ε4 genotypes and atorvastatin treatment in Brazilian individuals. The relationship of APOE genotypes and plasma lipids and atorvastatin response was also tested in this population. APOE ε2/ε3/ε4 and plasma lipids were evaluated in 181 normolipidemic (NL) and 181 hypercholesterolemic (HC) subjects. HC individuals with indication for lowering-cholesterol treatment (n = 141) were treated with atorvastatin (10 mg/day/4-weeks). APOE genotypes and APOE mRNA in peripheral blood mononuclear cells (PBMC) were analyzed by TaqMan real time PCR. HC had lower APOE expression than NL group (p < 0.05) and individuals with low APOE expression showed higher plasma total and LDL cholesterol and apoB, as well as higher apoAI (p < 0.05). Individuals carrying ε2 allele have reduced risk for hypercholesterolemia (OR: 0.27, 95% I.C.: 0.08-0.85, p < 0.05) and NL ε2 carriers had lower total and LDL cholesterol and apoB levels, and higher HDL cholesterol than non-carriers (p < 0.05). APOE genotypes did not affect APOE expression and atorvastatin response. Atorvastatin treatment do not modify APOE expression, however those individuals without LDL cholesterol goal achievement after atorvastatin treatment according to the IV Brazilian Guidelines for Dyslipidemia and Atherosclerosis Prevention had lower APOE expression than patients with desirable response after the treatment (p < 0.05). APOE expression in PBMC is modulated by hypercholesterolemia and the APOE mRNA level regulates the plasma lipid profile. Moreover the expression profile is not modulated neither by atorvastatin nor APOE genotypes. In our population, APOE ε2 allele confers protection against hypercholesterolemia and a less atherogenic lipid profile. Moreover, low APOE expression after treatment of patients with poor response suggests a possible role of APOE level in atorvastatin response.


The influence of polymer architecture on in vitro pDNA transfection.
In 2012, the first gene therapy agent was approved by the Europe Medicines Agency leading to increased interest in this research field. Beside viruses, non-viral agents based on lipids or polymers represent aspiring alternatives to deliver the genetic material. Different hurdles have to be overcome depending on the kind of nucleic acid used, where plasmid DNA (pDNA) and small interfering RNA represent the common ones. The main challenge for transfection agents, in particular for pDNA delivery, is the transfer to the cell and into the cell nuclei. Within the group of transfection vesicles, cationic polymers show promising features and variability, as they can be synthesized with tailor-made physical and chemical properties (architectures and functionalisation). In the field of polymer-based gene delivery, the tuning potential of polymers by using different architectures like graft and star-shaped polymers as well as self-assembled block copolymers is immense. In particular, in the last few years numerous new polymer designs showed enhanced transfection properties in combination with good biocompatibility. Furthermore, new insights into the transfection mechanism demonstrated the continuous progress in this field. Polymer architecture influences the polyplex characteristics and the latter has an impact on the transfection mechanism, e.g. the interaction with the cellular membrane depends on the polyplex shape. Moreover, polyplex dissociation can be easily influenced by the polymer chemistry, thus biodegradable linkers lead to well suited polymers with reduced toxicity and high delivery potential, and are also promising for in vivo applications. This review focuses on the influence of polymer architectures for pDNA transfection in vitro, showing recent developments and insights. The theoretical background concerning the biological challenges for cationic polymers and the impact of graft- or star-shaped architectures as well as self-assembled structures will be presented in detail.


Positron emission tomography imaging of brain tumors.
A wide variety of metabolic features of brain tumors can be imaged using PET, including glucose metabolism, blood flow, oxygen consumption, amino acid metabolism, and lipid synthesis. Currently, FDG is the most widely available PET tracer for body imaging and brain imaging. Malignant brain tumors, like many other soft tissue tumors, show increased glucose metabolism, which is reflected on FDG-PET imaging. FDG-PET imaging of brain tumors provides information on tumor grade and prognosis. Compared with other organ systems, FDG-PET imaging of the brain presents unique challenges because of the high background glucose metabolism of normal gray matter structures. Coregistration of the MRI (or CT) and FDG-PET images is essential for accurate evaluation of brain tumors and is performed routinely at the authors' institution. The heterogeneous nature of gliomas can result in significant sampling errors when patients are biopsied for primary tumor diagnosis or recurrent disease. FDG-PET can be used to define the most metabolically active targets for stereotactic biopsy. This in turn can improve diagnostic accuracy and reduce the number of biopsy samples required. FDG-PET is also useful for evaluating residual or recurrent tumor following therapy, and can be used to survey patients with low-grade brain tumors for evidence of degeneration into high-grade malignancy. In the case of suspected tumor recurrence or progression, PET can aid in defining appropriate targets for biopsy. One limitation of FDG-PET is the occasional inability to distinguish radiation necrosis from recurrent high-grade tumor. A second limitation is that FDG-PET is less sensitive than contrast-enhanced MRI for detecting intracranial metastases, and it is the authors' experience that brain studies should not be included as part of routine whole-body PET studies. Other tracers, such as 11C-methionine and FCH, also avidly accumulate in brain tumors and have the advantage of low background cortical activity. The relationship between degree of uptake of these agents and tumor grade is not established. These tracers may be useful in specific clinical situations, however, such as tumor localization for treatment planning or evaluation of low-grade tumors.


Population Genetics of Hirsutella rhossiliensis, a Dominant Parasite of Cyst Nematode Juveniles on a Continental Scale.
Hirsutella rhossiliensis is a parasite of juvenile nematodes, effective against a diversity of plant-parasitic nematodes. Its global distribution on various nematode hosts and its genetic variation for several geographic regions have been reported, while the global population genetic structure and factors underlying patterns of genetic variation of H. rhossiliensis are unclear. In this study, 87 H. rhossiliensis strains from five nematode species (Globodera sp., Criconemella xenoplax, Rotylenchus robustus, Heterodera schachtii, and Heterodera glycines) in Europe, the United States, and China were investigated by multilocus sequence analyses. A total of 280 variable sites (frequency, 0.6%) at eight loci and six clustering in high accordance with geographic populations or host nematode-associated populations were identified. Although H. rhossiliensis is currently recognized as an asexual fungus, recombination events were frequently detected. In addition, significant genetic isolation by geography and nematode hosts was revealed. Overall, our analyses showed that recombination, geographic isolation, and nematode host adaptation have played significant roles in the evolutionary history of H. rhossiliensis IMPORTANCE: H. rhossiliensis has great potential for use as a biocontrol agent to control nematodes in a sustainable manner as an endoparasitic fungus. Therefore, this study has important implications for the use of H. rhossiliensis as a biocontrol agent and provides interesting insights into the biology of this species.


Omega Rhodopsins: A Versatile Class of Microbial Rhodopsins.
Microbial rhodopsins are a superfamily of photoactive membrane proteins with covalently bound retinal cofactor. Isomerization of the retinal chromophore upon absorption of a photon triggers conformational changes of the protein to function as ion pumps or sensors. After the discovery of proteorhodopsin in an uncultivated γ-proteobacterium, light-activated proton pumps have been widely detected among marine bacteria and, together with chlorophyll-based photosynthesis, are considered as an important axis responsible for primary production in the biosphere. Rhodopsins and related proteins show a high level of phylogenetic diversity; we focus on a specific class of bacterial rhodopsins containing the 3 omega motif. This motif forms a stack of three nonconsecutive aromatic amino acids that correlates with the B-C loop orientation, and is shared among the phylogenetically close ion pumps such as the NDQ motif-containing sodium-pumping rhodopsin, the NTQ motif-containing chloride-pumping rhodopsin, and some proton-pumping rhodopsins including xanthorhodopsin. Here, we reviewed the recent research progress on these omega rhodopsins, and speculated on their evolutionary origin of functional diversity..


Therapeutic effects of cranial osteopathic manipulative medicine: a systematic review.
Cranial osteopathic manipulative medicine (OMM) involves the manipulation of the primary respiratory mechanism to improve structure and function in children and adults. To identify and critically evaluate the literature regarding the clinical efficacy of cranial OMM. The clinical keywords "cranial manipulation" OR "osteopathy in the cranial field" OR "cranial osteopathy" OR "craniosacral technique" were searched in the following electronic databases: EMBASE, MEDLINE In-Process & Other Non-Indexed Citations, The Cochrane Central Register of Controlled Trials, CINAHL (Cumulative Index to Nursing and Allied Health Literature), and AMED (Alternative Medicine). Searches were conducted in April 2011 with no date restriction for when the studies were completed. Randomized controlled trials and observational studies that measured the effectiveness of cranial OMM on humans were included in the study. Exclusion criteria included non-English language articles, studies not relevant to cranial OMM, animal studies, and studies in which there was no clear indication of the use of cranial OMM. Studies that described the use of cranial OMM with other treatment modalities and that did not perform subgroup analysis were also excluded. The present study did not have criteria regarding type of disease. Outcome measures on pain, sleep, quality of life, motor function, and autonomic nervous system function were extracted. The methodological quality of the trials was assessed using the Downs and Black checklist. Of the 8 studies that met the inclusion criteria, 7 were randomized controlled trials and 1 was an observational study. A range of cranial OMM techniques used for the management of a variety of conditions were identified in the included studies. Positive clinical outcomes were reported for pain reduction, change in autonomic nervous system function, and improvement of sleeping patterns. Methodological Downs and Black quality scores ranged from 14 to 23 points out of a maximum of 27 points (overall median score, 16). The currently available evidence on the clinical efficacy of cranial OMM is heterogeneous and insufficient to draw definitive conclusions. Because of the moderate methodological quality of the studies and scarcity of available data, further research into this area is needed.


Frequency and nature of coroners' recommendations from injury-related deaths among nursing home residents: a retrospective national cross-sectional study.
Coroners are tasked with the investigation of unnatural and unexpected deaths. In Australia, the coroner's role also includes making recommendations for promoting interventions to improve public safety. However, the coroners' role in public health and safety in the aged care setting is an underexplored area of research. To describe the frequency and nature of coroners' recommendations for prevention of harm from injury-related deaths among nursing home residents in Australia. Accredited nursing homes in Australia. Nursing home residents whose deaths resulted from external causes occurring between 1 July 2000 and 31 December 2013 and notified to a coroner. Coroners' recommendations were identified and extracted from the National Coronial Information System. Descriptive statistical techniques were used to calculate the frequency and proportion of recommendations made. The nature of recommendations was quantified using a method comprising seven elements derived from internationally accepted and applied public health conceptual models of mortality causation and prevention. Coroners made recommendations about the prevention of harm in 53 of the 3289 (1.6%) external cause deaths of nursing home residents. Recommendations were most frequently made for deaths resulting from falls; however, the rate of recommendations per 1000 deaths was highest for thermal mechanisms and complications of clinical care. Most recommendations described the 'countermeasure' element, but rarely specified a timeframe for implementation. Coroners' recommendations need to be further enhanced in the age care setting. The development of national and international guidelines on best practice in the formulation of effective recommendations should be undertaken.


Targeting ALK: Precision Medicine Takes on Drug Resistance.
Anaplastic lymphoma kinase (ALK) is a validated molecular target in several ALK-rearranged malignancies, including non-small cell lung cancer. However, the clinical benefit of targeting ALK using tyrosine kinase inhibitors (TKI) is almost universally limited by the emergence of drug resistance. Diverse mechanisms of resistance to ALK TKIs have now been discovered, and these basic mechanisms are informing the development of novel therapeutic strategies to overcome resistance in the clinic. In this review, we summarize the current successes and challenges of targeting ALK. Effective long-term treatment of ALK-rearranged cancers requires a mechanistic understanding of resistance to ALK TKIs so that rational therapies can be selected to combat resistance. This review underscores the importance of serial biopsies in capturing the dynamic therapeutic vulnerabilities within a patient's tumor and offers a perspective into the complexity of on-target and off-target ALK TKI resistance mechanisms. Therapeutic strategies that can successfully overcome, and potentially prevent, these resistance mechanisms will have the greatest impact on patient outcome. Cancer Discov; 7(2); 137-55. ©2017 AACR.


Human papillomavirus-associated oropharyngeal cancer: review of current evidence and management.
Oropharyngeal cancer (opc) has become the leading site for human papillomavirus (hpv)-associated cancers in humans. It is an epidemic that remains relatively unfamiliar to most physicians, potentially delaying diagnosis and treatment. Traditionally, cancers involving the head and neck have occurred in smokers and in those with a significant alcohol history. Typically, hpv-positive opc presents in a younger, healthier population with a different set of risk factors and good prognosis for survival. However, many head-and-neck cancer patients, including those with hpv-positive disease, develop lifelong disabilities because of the morbid nature of their treatments, and those patients have the highest level of unmet needs in studies spanning cancer sites. Knowledge of this epidemic, a high index of suspicion, and an understanding of how the tumours present in clinical practice can help physicians to make an early diagnosis, thus sparing the patient significant morbidity from treatments associated with more advanced disease stages. Furthermore, recognizing that these patients have distinct psychosocial needs and implementing a collaborative team approach is critical to providing optimal care and improving quality of life in the survivorship period.


Intraarterial Hepatic SPECT/CT Imaging Using 99mTc-Macroaggregated Albumin in Preparation for Radioembolization.
Current standard practice for radioembolization treatment planning makes use of nuclear medicine imaging (NMI) of (99m)Tc-macroaggregated albumin ((99m)Tc-MAA) arterial distributions for the assessment of lung shunting and extrahepatic uptake. Our aim was to retrospectively compare NMI with mapping angiography in the detection and localization of extrahepatic (99m)Tc-MAA and to evaluate the typical and atypical findings of NMI in association with catheter placement. One hundred seventy-four patients underwent diagnostic angiography in preparation for radioembolization. (99m)Tc-MAA was administered to the liver via a microcatheter positioned in the desired hepatic artery. Planar scintigraphy imaging followed by SPECT/CT imaging was obtained within 2 h. All images were reviewed for hepatic and extrahepatic (99m)Tc-MAA deposition and compared with the mapping angiogram. Intrahepatic lobe shunting was present on NMI in only 2.9% of the cases but was present in 62.5% of the patients with portal vein thrombosis. Extrahepatic distributions included lungs (100%), the gallbladder (49%) if present, and locations involving hepaticoenteric arterial anatomy recognized on angiograms (16%). Free pertechnetate was identified on 38% of the nuclear medicine images. Three percent of nuclear medicine images showed alternative findings such as a thyroid nodule or metallic artifact. Patients being considered for radioembolization should undergo both angiography and scintigraphy for the assessment of hepaticoenteric arterial anatomy, hepatopulmonary shunting, and appropriate dosimetry considerations. Knowledge of the expected distribution of (99m)Tc-MAA with normal variants and potential nontarget delivery to adjacent structures is critical in improving clinical outcomes.


Broadband scattering properties of articular cartilage zones and their relationship with the heterogenous structure of articular cartilage extracellular matrix.
Articular cartilage exhibits a zonal architecture, comprising three distinct zones: superficial, middle, and deep. Collagen fibers, being the main solid constituent of articular cartilage, exhibit unique angular and size distribution in articular cartilage zones. There is a gap in knowledge on how the unique properties of collagen fibers across articular cartilage zones affect the scattering properties of the tissue. This study hypothesizes that the structural properties of articular cartilage zones affect its scattering parameters. We provide scattering coefficient and scattering anisotropy factor of articular cartilage zones in the spectral band of 400 to 1400 nm. We enumerate the differences and similarities of the scattering properties of articular cartilage zones and provide reasoning for these observations. We utilized collimated transmittance and integrating sphere measurements to estimate the scattering coefficients of bovine articular cartilage zones and bulk tissue. We used the relationship between the scattering coefficients to estimate the scattering anisotropy factor. Polarized light microscopy was applied to estimate the depth-wise angular distribution of collagen fibers in bovine articular cartilage. We report that the Rayleigh scatterers contribution to the scattering coefficients, the intensity of the light scattered by the Rayleigh and Mie scatterers, and the angular distribution of collagen fibers across tissue depth are the key parameters that affect the scattering properties of articular cartilage zones and bulk tissue. Our results indicate that in the short visible region, the superficial and middle zones of articular cartilage affect the scattering properties of the tissue, whereas in the far visible and near-infrared regions, the articular cartilage deep zone determines articular cartilage scattering properties. This study provides scattering properties of articular cartilage zones. Such findings support future research to utilize optical simulation to estimate the penetration depth, depth-origin, and pathlength of light in articular cartilage for optical diagnosis of the tissue.


Amiodarone-induced block of sodium current in isolated cardiac cells.
Sodium current (INa) block by amiodarone (AMI) was investigated in isolated single Purkinje and ventricular myocardial cells using the single suction-pipette voltage-clamp technique. AMI produced marked resting block that was enhanced at low holding potentials, findings consistent with a shift in the steady-state INa availability curve to more negative potentials (-16 +/- 3 mV). Resting block was not associated with any change in the time course of INa decay during a depolarizing clamp step. AMI also produced use-dependent block in conjunction with increases in rate (0.5-5.0 Hz) and pulse duration (2-200 msec). These changes are consistent with a slowing of the recovery from inactivation of the sodium channel. Brief depolarizing pulses produced little use-dependent block, suggesting that the onset of drug-induced block is slow. Thus, AMI blocks INa and shifts the availability curve in isolated myocytes, both of which contribute to the net tonic block. The results suggest that both rested state and inactivated state sodium channel block are factors in AMI's antiarrhythmic efficacy.


The Potential Importance of Detection of Neovascular Age-Related Macular Degeneration When Visual Acuity Is Relatively Good.
The advent of anti-vascular endothelial growth factor treatment has changed the prognosis for patients with neovascular age-related macular degeneration (nvAMD). The ability to stabilize or improve vision with these treatments is a major step in enabling patients to continue to function at the highest possible level. Many studies have demonstrated that the better the visual acuity (VA) is at the time of treatment initiation, the higher the likelihood that VA will be better during at least the following 2 years; as such, detection of nvAMD when VA is relatively good is important. Data on the VA of patients with intermediate AMD and VA at the time of nvAMD diagnosis suggest that patients are typically losing an average of 3 to 5 lines of vision and possibly more between the time that intermediate AMD progresses to nvAMD and the diagnosis of nvAMD is made. The average patient may have nvAMD for 6 to 12 months before diagnosis and treatment initiation. Current efforts in management of nvAMD are primarily aimed at optimizing anti-vascular endothelial growth factor treatments that have the potential to improve VA outcomes by a magnitude of letters. Additional tools or other efforts to identify patients with nvAMD before substantial vision loss has occurred may reduce the amount of visual loss sustained with anti-vascular endothelial growth factor therapy, and have the potential to improve VA outcomes substantially.


Control over surface DNA density on gold nanoparticles allows selective and sensitive detection of mercury(II).
We have developed a new highly selective and sensitive technique for the detection of Hg(2+) using DNA-functionalized gold nanoparticles (Au NPs) and OliGreen. This system is the first that allows the detection of Hg(2+) based on the release of DNA molecules, induced by conformational changes on Au NP surfaces, and its sensitivity is highly dependent upon surface DNA density. When Hg(2+) ions interact with the thymidine units of the DNA molecules bound to the Au NPs through Au-S bonds, the conformations of these DNA derivatives change from linear to hairpin structures, causing the release of some of the DNA molecules from the surface of the Au NPs into the bulk solution to react with OliGreen. The fluorescence of OliGreen-DNA complexes increased with increasing concentration of Hg(2+), and Hg(2+) could be detected at concentrations as low as 25 nM. A linear correlation existed between the fluorescence intensity and the concentration of Hg(2+) over the range 0.05-2.5 microM (R(2) = 0.98). This simple and cost-effective probe was applied to determine the spiked Hg(2+) in the pond samples; the recoveries (96-102%) suggested low matrix interference and high sensitivity.


Beta diversity as the variance of community data: dissimilarity coefficients and partitioning.
Beta diversity can be measured in different ways. Among these, the total variance of the community data table Y can be used as an estimate of beta diversity. We show how the total variance of Y can be calculated either directly or through a dissimilarity matrix obtained using any dissimilarity index deemed appropriate for pairwise comparisons of community composition data. We addressed the question of which index to use by coding 16 indices using 14 properties that are necessary for beta assessment, comparability among data sets, sampling issues and ordination. Our comparison analysis classified the coefficients under study into five types, three of which are appropriate for beta diversity assessment. Our approach links the concept of beta diversity with the analysis of community data by commonly used methods like ordination and anova. Total beta can be partitioned into Species Contributions (SCBD: degree of variation of individual species across the study area) and Local Contributions (LCBD: comparative indicators of the ecological uniqueness of the sites) to Beta Diversity. Moreover, total beta can be broken up into within- and among-group components by manova, into orthogonal axes by ordination, into spatial scales by eigenfunction analysis or among explanatory data sets by variation partitioning.


Nelarabine, etoposide, and cyclophosphamide in relapsed pediatric T-acute lymphoblastic leukemia and T-lymphoblastic lymphoma (study T2008-002 NECTAR).
Children with relapse of T-cell acute lymphoblastic leukemia (T-ALL) or lymphoblastic lymphoma (T-LBL) have a dismal prognosis, largely due to difficulty attaining second remission. We hypothesized that adding etoposide and cyclophosphamide to the nucleoside analog nelarabine could improve response rates over single-agent nelarabine for relapsed T-ALL and T-LBL. This phase I dose-escalation trial's primary objective was to evaluate the dose and safety of nelarabine given in combination with etoposide at 100 mg/m2 /day and cyclophosphamide at 330-400 mg/m2 /day, each for 5 consecutive days in children with either T-ALL (13 patients) or T-LBL (10 patients). Twenty-three patients were treated at three dose levels; 21 were evaluable for dose-limiting toxicities (DLT) and response. The recommended phase II doses (RP2D) for this regimen, when given daily ×5 every 3 weeks, were nelarabine 650 mg/m2 /day, etoposide 100 mg/m2 /day, and cyclophosphamide 400 mg/m2 /day. DLTs included peripheral motor and sensory neuropathies. An expansion cohort to evaluate responses at the RP2D was terminated early due to slow accrual. The overall best response rate was 38% (8/21), with 33% (4/12) responses in the T-ALL cohort and 44% (4/9) responses in the T-LBL cohort. These response rates are comparable to those seen with single-agent nelarabine in this setting. These data suggest that the addition of cyclophosphamide and etoposide to nelarabine does not increase the incidence of neurologic toxicities or the response rate beyond that obtained with single-agent nelarabine in children with first relapse of T-ALL and T-LBL.


Memory function in normal aging.
We evaluated memory function in a group of 161 community-dwelling, cognitively normal individuals aged 62 to 100 years recruited as part of the Mayo Clinic Alzheimer's Disease Patient Registry. We used the Free and Cued Selective Reminding Test and the Rey Auditory Verbal Learning Test to evaluate two aspects of memory function thought to be sensitive to the effects of aging: learning (acquisition) and delayed recall (forgetting). The results were quite consistent and demonstrated that learning or acquisition performance declines uniformly with increasing age but is not related to education. Delayed recall or forgetting, however, remained relatively stable across age when adjusted for the amount of material initially learned. These findings are relevant for assessing normal memory function relative to the early impairments found in dementia and form a baseline against which memory performance can be assessed by the clinician. In particular, suspicion regarding a disorder of brain function affecting memory processes should be raised if learning performance declines more rapidly than expected or if delayed recall is impaired to any significant extent.


Dielectric Barrier Discharge Applicator for Heating Carbon Nanotube-Loaded Interfaces and Enhancing 3D-Printed Bond Strength.
Material extrusion (ME) 3D printing is a revolutionary technique for manufacturing thermoplastic parts; however, the printed parts typically suffer from poor interlayer bonding, which causes weak tensile strength in the build direction. Many methods have been proposed to address the mechanical deficiencies of 3D-printed parts, but most fall short of a production-ready solution. Here we report the use of a dielectric barrier discharge (DBD) plasma electrode mounted concentrically around the nozzle of an ME 3D printer for in situ welding of thermoplastic parts. This is the first report of a DBD being used as a non-contact means to induce Joule heating in resistive composite materials. The polymer welding process is accomplished by coupling the DBD with the carbon nanotube-loaded interfaces between the 3D-printed layers. The current passing through the part results in rapid resistive heating of the nanotubes and thermal welding of the interfaces. We show that parts printed with this method have isotropic strength and are equivalent to their injection-molded counterparts.


Absorption of foliar-applied Zn fertilizers by trichomes in soybean and tomato.
The present study investigated the role of trichomes in absorption of foliar-applied zinc fertilizers in soybean and tomato. Using synchrotron-based X-ray fluorescence microscopy for in situ analyses of hydrated leaves, we found that upon foliar application of ZnSO4, Zn accumulated within 15 min in some non-glandular trichomes in soybean, but not in tomato. However, analyses of cross-sections of soybean leaves did not show any marked accumulation of Zn in tissues surrounding trichomes. Furthermore, when near-isogenic lines of soybean differing 10-fold in trichome density were used to compare Zn absorption, it was found that foliar Zn absorption was not related to trichome density. Therefore, it is suggested that trichomes are not part of the primary pathway through which foliar-applied Zn moves across the leaf surface in soybean and tomato. However, this does not preclude trichomes being important in other plant species, as they are known to be highly diverse. We also compared the absorption of Zn when supplied as either ZnSO4, nano-ZnO, or bulk-ZnO, and found that absorption from ZnSO4 was about 10-fold higher than from nano- and bulk-ZnO, suggesting that it was mainly absorbed as soluble Zn. This study improves our understanding of the absorption of foliar-applied nutrients.


Adsorption of Ten Microcystin Congeners to Common Laboratory-Ware Is Solvent and Surface Dependent.
Cyanobacteria can produce heptapetides called microcystins (MC) which are harmful to humans due to their ability to inhibit cellular protein phosphatases. Quantitation of these toxins can be hampered by their adsorption to common laboratory-ware during sample processing and analysis. Because of their structural diversity (>100 congeners) and different physico-chemical properties, they vary in their adsorption to surfaces. In this study, the adsorption of ten different MC congeners (encompassing non-arginated to doubly-arginated congeners) to common laboratory-ware was assessed using different solvent combinations. Sample handling steps were mimicked with glass and polypropylene pipettes and vials with increasing methanol concentrations at two pH levels, before analysis by liquid chromatography-tandem mass spectrometry. We demonstrated that MC adsorb to polypropylene surfaces irrespective of pH. After eight successive pipet actions using polypropylene tips ca. 20% of the MC were lost to the surface material, which increased to 25%-40% when solutions were acidified. The observed loss was alleviated by changing the methanol (MeOH) concentration in the final solvent. The required MeOH concentration varied depending on which congener was present. Microcystins only adsorbed to glass pipettes (loss up to 30% after eight pipet actions) when in acidified aqueous solutions. The latter appeared largely dependent on the presence of ionizable groups, such as arginine residues.


Radiologic analysis of femoral acetabular impingement: from radiography to MRI.
Femoral acetabular impingement is a set of morphologic abnormalities that are considered to be a major cause of degenerative disease in the hip joint. Early changes are already present in adolescence when it is the pediatric radiologist who must assess current damage with the aim of averting progression to more severe and debilitating osteoarthritis. A multimodality approach is used for diagnosis, that includes conventional radiography and CT to assess the osseous structures. MR arthrography is the primary advanced imaging modality for assessment of morphologic changes as well as injuries of the labrum and articular cartilage. Details of radiologic imaging are offered to guide the radiologist and provide an avenue for the accurate description of the osseous and articular alterations and injury.


Recent Advances in the Management of Major Postpartum Haemorrhage - A Review.
Postpartum haemorrhage (PPH) is a leading cause of maternal mortality and morbidity worldwide and 75-90% of these haemorrhage results from uterine atony. Delayed and substandard obstetrics care can kill a woman within hours of Major Obstetric Haemorrhage (MOH). Prenatal identification of at risk women, prompt assessment of blood loss, effective management and involvement of multidisciplinary teams is of utmost importance to save the lives of these women. However, even with the best prenatal care, PPH occurs, it can occur without any risk factors. The first step in management is achieving haemodynamic stability, second being arrest of bleeding, both are done simultaneously. Cases of refractory PPH is managed by postpartum hysterectomy which results in complete inability in hosting a future pregnancy, a psychological impact and risk of intra operative surgical morbidities. This review discusses the current evidence based management of PPH, existing controversies in transfusion of blood and blood products and newer advances in this field. It was conducted by searching the English language medical literature using Medline (1994-2015). The current scenario in developing countries mandates research on newer and practicable strategies to tackle PPH which can be implemented effectively and have an upper edge over the existing practices in the management of PPH.


The influence of pH and nicotine concentration in oral moist snuff on mucosal changes and salivary pH in Swedish snuff users.
The use of Swedish oral moist snuff is a widespread habit in Sweden. In 1999, 25% of the adult male population and 3.1% of the female population were users of snuff. The aim of the present study was to evaluate how variations in pH and nicotine concentrations of snuff affect the oral mucosa, clinically and histologically, salivary pH and daily nicotine intake in 20 habitual users of loose snuff. The subjects were studied during use of their usual brand, after 12 weeks use of a snuff with lower pH and after another 12 weeks use of a snuff with both lower pH and lower nicotine concentration. Consumption data, oral soft tissue changes, salivary pH and nicotine intake were measured. Further, biopsies were taken from the central part of the clinically observed lesions and histological changes were analysed. The subjects maintained their snuff consumption during the observation periods. The average salivary pH was higher during snuff use than in the morning. Further, it was higher shortly after the snuff was removed than during snuff use. After having switched to the snuff with both lower pH and lower nicotine concentrations, they showed a significant reduction in daily nicotine intake and developed significantly less pronounced clinical and histological changes. These results indicate that nicotine is one of the substances in snuff that has a biological effect on the oral mucosa. However, there also seems to be a synergistic effect between the pH and nicotine concentration in the snuff.


Normal Fetal Long Bone Length from Computed Tomography: Potential Value in the Prenatal Evaluation of Skeletal Dysplasias.
The prenatal evaluation of skeletal dysplasias is challenging. Ultrasound (US) has limited sensitivity in evaluating these abnormalities. Low-dose fetal computed tomography (CT) is a helpful adjunct in evaluating the fetus with severe abnormalities, but its use is limited by lack of data on normal fetal bone length. To obtain CT measurements of fetal long bones and to compare published US normative bone length measures to examine their similarity. A retrospective search of a radiographic database for gravid patients with CT of the abdomen/pelvis for acute maternal abdomen was performed. Fetal long bone measurements were plotted against gestational age (GA)/standard US measurements. Regression coefficients were calculated (p values <0.05 were considered significant). Bland-Altman plots were constructed to identify any systematic difference between CT and US bone length measures. Twenty-five cases met the inclusion criterion of fetal GA 16-39 weeks. The length of long bones increased with GA, with strong R2 values for each measurement. Plots of CT measurements against published 50th percentile sonographic values for age also showed strong and significant correlations, suggesting that values may be used interchangeably, a conclusion further substantiated by the Bland-Altman analyses. A CT database for fetal long bone length is presented which can be used to evaluate the fetus with abnormal bones. Additionally we have established a strong correlation between measured fetal CT long bone values and the published 50th percentile normal values for US, which allows the interpreting radiologist to extrapolate CT measurements to US values in the evaluation of fetal bone length.


[Preoperative determination of the boundaries of soft tissue necrosis in patients with diabetes mellitus].
The authors examined 34 patients with diabetes mellitus and purulent diseases of the soft tissues and 40 persons who formed the control group. The study of local microcirculation in the purulent focus, in a purulent and closed wound by determining the oxygen tension in capillary blood (pO2) established by using an noninvasive method that a real possibility for determining the exact boundaries of tissue necrosis has appeared. This makes it possible to excise optimally the necrotic tissues and choose the techniques of applying primary, primary-delayed or early secondary sutures. In addition, the study of the dynamics of pO2 changes allows the course of the wound process to be followed up and, consequently, possible suppuration of the wound to be predicted.


[The urbanization and demographic structure of population].
Analyzes the effects of urbanization on the demographic structure of population. An increase in the degree of urbanization was found to be paralleled by an increase of the age qualification and of the migration activity of the population. The national composition of the population does not depend on urbanization.


How to isolate a ready-to-use adipose-derived stem cells pellet for clinical application.
Adipose-derived stem cells (ASCs) are multipotent mesenchymal stem cells (MSCs) that show definitive stem cell characteristics such as plastic adherence in culture, ability to maintain multipotency upon in vitro expansion, and self-renewal capacity. ASCs are particularly promising for use in regenerative medicine because they can be harvested easily from adipose tissue by standard liposuction, with minimal donor site morbidity. Since ASCs do not necessitate ex vivo expansion to obtain clinically significant cell numbers, it is critical to identify a standardized method that maximizes the number of ASCs collected. Based on current literature, there is no standardized method to isolate ASCs for clinical application. Furthermore, clinical studies involving ASCs often show inconsistencies in the reported results. Such studies often use research-derived isolation protocols, which are complex, time-consuming, and involve the use of chemical and animal-derived reagents. In this paper, we present an in-depth review of the available data on ASC isolation protocols. Moreover, we describe our isolation protocol that allows the collection of a ready-to-use ASC pellet for clinical application.


Dynamics and logic of promoter melting.
Despite significant progress in our understanding of promoter melting dynamics, underlying principles of the process remain elusive, with opposing views on key aspects held by many in the field. Here, I discuss the mechanistic logic behind the interplay of thermal and deterministic forces acting to create transcriptionally competent promoter complexes.


Education can improve the negative perception of a threatened long-lived scavenging bird, the Andean condor.
Human-wildlife conflicts currently represent one of the main conservation problems for wildlife species around the world. Vultures have serious conservation concerns, many of which are related to people's adverse perception about them due to the belief that they prey on livestock. Our aim was to assess local perception and the factors influencing people's perception of the largest scavenging bird in South America, the Andean condor. For this, we interviewed 112 people from Valle Fértil, San Juan province, a rural area of central west Argentina. Overall, people in the area mostly have an elementary education, and their most important activity is livestock rearing. The results showed that, in general, most people perceive the Andean condor as an injurious species and, in fact, some people recognize that they still kill condors. We identified two major factors that affect this perception, the education level of villagers and their relationship with livestock ranching. Our study suggests that conservation of condors and other similar scavengers depends on education programs designed to change the negative perception people have about them. Such programs should be particularly focused on ranchers since they are the ones who have the worst perception of these scavengers. We suggest that highlighting the central ecological role of scavengers and recovering their cultural value would be fundamental to reverse their persecution and their negative perception by people.


Kleihauer-betke testing is important in all cases of maternal trauma.
In maternal trauma, the Kleihauer-Betke (KB) test has traditionally been used to detect transplacental hemorrhage (TPH), so that Rh-negative women could receive appropriate Rh immune prophylaxis. Reasoning that the magnitude of TPH would reflect uterine injury, we evaluated Kleihauer-Betke testing as an independent predictor of preterm labor (PTL) after maternal trauma. Admissions to the Shock Trauma Center, University of Maryland, from January 1996 to January 2002, were reviewed. Of 30,362 trauma patients admitted, 166 were pregnant, and 93 of these underwent electronic fetal monitoring. Their records were abstracted for demographics, injury type, three separate trauma scores, documented uterine contractions, PTL (contractions with progressive cervical change), and serious perinatal complications. In 71 cases, transplacental hemorrhage was assessed by maternal KB test. TPH, defined as KB-positive for greater than 0.01 mL of fetal blood in the maternal circulation, occurred in 46 women. Forty-four had documented contractions (25 had overt PTL) and 2 had no contractions. In 25 women with a negative KB test, none had uterine contractions. All patients with contractions or PTL had positive KB tests. By logistic regression, KB test result was the single risk factor associated with PTL (p < 0.001; likelihood ratio, 20.8 for positive KB test). Compared with other sites, abdominal trauma was associated more often with uterine contractions (p < 0.001), PTL (p = 0.001), and a positive KB test (p < 0.001, chi). None of the trauma scoring systems predicted PTL. Kleihauer-Betke testing accurately predicts the risk of preterm labor after maternal trauma. Clinical assessment does not. With a negative KB test, posttrauma electronic fetal monitoring duration may be limited safely. With a positive KB test, the significant risk of PTL mandates detailed monitoring. KB testing has important advantages to all maternal trauma victims, regardless of Rh status.


Temporal and spatial expression of tissue inhibitors of metalloproteinases during the natural ovulatory cycle of the mouse.
The role of tissue inhibitors of metalloproteinases (TIMPs) in ovarian function has primarily been documented by studies that utilize hormone-primed animals. In this study, our objectives were to elucidate the spatiotemporal expression of individual TIMP genes during the natural ovulatory cycle, and to correlate these with specific biological events. Two models of spontaneous ovulation used were the murine estrous cycle and the first ovulation postpartum. Ovaries were collected from mice at diestrus, estrus, and metestrus, or at early and late proestrus, and from pregnant females on Days 17 and 18 of gestation (D17, D18) and within 24 or 48 h postpartum (PP1, PP2). We observed that TIMP-1 mRNA was elevated at early proestrus and D18 and was maximal at late proestrus and PP1. The TIMP-3 pattern was distinct from that of TIMP-1, maximal expression occurring at early proestrus and D17 and D18. In both models, TIMP-2 mRNA remained constant and at very low levels throughout ovulation. In situ hybridization localized TIMP-1 mRNA to the corpus luteum at D18 and PP1, and to oocytes at specific stages of follicular development. Expression of TIMP-1 in granulosa and thecal cells was not observed at any stage. Demonstrating a distinct distribution, TIMP-3 mRNA was localized to oocytes, thecal and granulosa cells of small and large follicles, and corpora lutea only at D17. These data suggest specific hormonal regulation of individual TIMP gene expression in the ovary associated with distinct physiological functions. We propose that in the natural ovulatory process, TIMP-1 is probably a factor that regulates corpus luteum regression while TIMP-3 is important in maintaining the structural integrity of the corpus luteum.


In vivo imaging of cerebral hemodynamics using high-frequency micro-ultrasound.
Assessment of cerebral vascular response is important in neuroscience research. Some imaging modalities that are commonly used to detect flow and/or vessel diameter changes in the brain include magnetic resonance imaging, positron emission tomography, and optical intrinsic signal imaging. Ultrasound has not typically been used to assess neurovascular response but recent advances in the technology have led to the development of micro-ultrasound systems with significant potential for this application. The state of the art in high frequency (15-50 MHz) micro-ultrasound is based on linear arrays specifically designed for small animal imaging. These systems can achieve axial resolution ranging from 30 to 200 microm. They are capable of quantifying brain hemodynamics in terms of red blood cell (RBC) velocity, flow, and vascular density in real time, up to 35 mm below the cortical surface, and can achieve temporal resolution of up to 1000 frames per second. This protocol describes imaging of the rat brain using various ultrasound imaging modes (power Doppler, color Doppler, pulsed-wave Doppler, and nonlinear contrast-enhanced imaging) to assess the state of cerebral microcirculation.


Phase transition of light on complex quantum networks.
Recent advances in quantum optics and atomic physics allow for an unprecedented level of control over light-matter interactions, which can be exploited to investigate new physical phenomena. In this work we are interested in the role played by the topology of quantum networks describing coupled optical cavities and local atomic degrees of freedom. In particular, using a mean-field approximation, we study the phase diagram of the Jaynes-Cummings-Hubbard model on complex networks topologies, and we characterize the transition between a Mott-like phase of localized polaritons and a superfluid phase. We found that, for complex topologies, the phase diagram is nontrivial and well defined in the thermodynamic limit only if the hopping coefficient scales like the inverse of the maximal eigenvalue of the adjacency matrix of the network. Furthermore we provide numerical evidences that, for some complex network topologies, this scaling implies an asymptotically vanishing hopping coefficient in the limit of large network sizes. The latter result suggests the interesting possibility of observing quantum phase transitions of light on complex quantum networks even with very small couplings between the optical cavities.


Bariatric surgery tourism hidden costs? How Canada is not doing its part in covering bariatric surgery under the Canada Health Act.
Many Canadians seek medical treatment outside our borders. Waiting times, rather than lack of expertise, are the number one culprit, and with globalization of health care, the number of patients who travel to obtain medical care will continue to rise. Though the provinces have covered the costs of complications from surgeries performed abroad for many years, complications from bariatric surgery performed abroad have been receiving negative attention. This commentary discusses associated costs and questions how the Canada Health Act should be covering bariatric procedures.


Gender and age influence the relationship between serum GH and IGF-I in patients with acromegaly.
In patients with acromegaly serum IGF-I is increasingly used as a marker of disease activity. As a result, the relationship between serum GH and IGF-I is of profound interest. Healthy females secrete three times more GH than males but have broadly similar serum IGF-I levels, and women with GH deficiency require 30-50% more exogenous GH to maintain the same serum IGF-I as GH-deficient men. In a selected cohort of patients with active acromegaly, studied off medical therapy using a single fasting serum GH and IGF-I measurement, we have reported previously that, for a given GH level, women have significantly lower circulating IGF-I. To evaluate the influence of age and gender on the relationship between serum GH and IGF-I in an unselected cohort of patients with acromegaly independent of disease control and medical therapy. Sixty (34 male) unselected patients with acromegaly (median age 51 years (range 24-81 years) attending a colonoscopy screening programme were studied. Forty-five had previously received pituitary radiotherapy. Patients had varying degrees of disease control and received medical therapy where appropriate. Mean serum GH was calculated from an eight-point day profile (n = 45) and values obtained during a 75-g oral glucose tolerance test (n = 15). Serum IGF-I, IGFBP-3 and acid-labile subunit were measured and the dependency of these factors on covariates such as log10 mean serum GH, sex, age and prior radiotherapy was assessed using regression techniques. The median calculated GH value was 4.7 mU/l (range 1-104). A significant linear association was observed between serum IGF-I and log10 mean serum GH for the cohort (R = 0.5, P < 0.0001). After simultaneous adjustment of the above covariates a significant difference in the relationship between mean serum GH and IGF-I was observed for males and females. On average, women had serum IGF-I levels 11.44 nmol/l lower than men with the same mean serum GH (P = 0.03, 95% CI 1.33-21.4 nmol/l). Age significantly influenced the relationship and for a given serum GH, IGF-I was estimated to fall by 0.37 nmol/l per year (P = 0.04, 95% CI 0.015-0.72). In keeping with previous observations of relative GH resistance in normal and GH-deficient females we have observed lower serum IGF-I levels for equivalent mean serum GH levels in females patients with acromegaly. This gender-dependent difference is independent of disease activity and the use of concomitant medical therapy. Additionally, we have demonstrated that for a given serum GH level, age significantly influences IGF-I concentrations in patients with acromegaly. These data have important implications for the use of serum IGF-I and GH as markers of disease activity in acromegaly.


Rapamycin and caspofungin show synergistic antifungal effects in caspofungin-susceptible and caspofungin-resistant Candida strains in vitro.
Caspofungin is an echinocandin antifungal agent that inhibits synthesis of glucan required for the fungal cell wall. Resistance is mediated by mutation of Fks1 glucan synthase, among which S645P is the most common resistance-associated polymorphism. Rapamycin is a macrolide that inhibits the mechanistic target of rapamycin (mTOR) protein kinase activity. This study investigated the interaction between rapamycin and caspofungin in inhibiting the growth of WT Candida albicans and Fks1 S645P mutant clinical isolate, and WT Candida lusitaniae and genetically engineered isogenic strain with Fks1 S645P mutation at equivalent position. Interactions between caspofungin and rapamycin were evaluated using the microdilution chequerboard method in liquid medium. The results were analysed using the Loewe additivity model (FIC index, FICI) and the Bliss independence model (response surface, RS, analysis). Synergy between rapamycin and caspofungin was shown for C. albicans and C. lusitaniae strains by RS analysis of the chequerboard tests. Synergy was observed in strains susceptible and resistant to caspofungin. Weak subinhibitory concentrations of rapamycin were sufficient to restore caspofungin susceptibility. We report here, for the first time, synergy between caspofungin and rapamycin in Candida species. Synergy was shown for strains susceptible and resistant to caspofungin. This study highlights the possible implication of the TOR pathway in sensing antifungal-mediated cell wall stress and in modulating the cellular response to echinocandins in Candida yeasts.


In vivo evidence for neuroplasticity in older adults.
Neuroplasticity can be conceptualized as an intrinsic property of the brain that enables modification of function and structure in response to environmental demands. Neuroplastic strengthening of synapses is believed to serve as a critical mechanism underlying learning, memory, and other cognitive functions. Ex vivo work investigating neuroplasticity has been done on hippocampal slices using high frequency stimulation. However, in vivo neuroplasticity in humans has been difficult to demonstrate. Recently, a long-term potentiation-like phenomenon, a form of neuroplastic change, was identified in young adults by differences in visual evoked potentials (VEPs) that were measured before and after tetanic visual stimulation (TVS). The current study investigated whether neuroplastic changes in the visual pathway can persist in older adults. Seventeen healthy subjects, 65 years and older, were recruited from the community. Subjects had a mean age of 77.4 years, mean education of 17 years, mean MMSE of 29.1, and demonstrated normal performance on neuropsychological tests. 1Hz checkerboard stimulation, presented randomly to the right or left visual hemi-field, was followed by 2min of 9Hz stimulation (TVS) to one hemi-field. After 2min of rest, 1Hz stimulation was repeated. Temporospatial principal component analysis was used to identify the N1b component of the VEPs, at lateral occipital locations, in response to 1Hz stimulation pre- and post-TVS. Results showed that the amplitude of factors representing the early and late N1b component was substantially larger after tetanic stimulation. These findings indicate that high frequency visual stimulation can enhance the N1b in cognitively high functioning old adults, suggesting that neuroplastic changes in visual pathways can continue into late life. Future studies are needed to determine the extent to which this marker of neuroplasticity is sustained over a longer period of time, and is influenced by age, cognitive status, and neurodegenerative disease.


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Cold burns are still exceptional in sub-Saharan Africa. However, the promotion of a wider use of domestic gas (commercial butane) has led to the growth of petrochemical industries. As a consequence, there is an increased risk of the occurrence of such burns (referred to as frostbite). We report a case of a cold burn injury caused by liquefied petroleum gas (LPG) in the workplace. The diagnosis of severity and management later complicated the local evolution. Compliance with safety measures in factories remains the main means of preventing this rarely seen type of burn.


A novel method of continuous cage-side monitoring of hyperthyroid cats treated with radio-iodine.
A continuous monitoring system (MGP DM2000X) was assessed for monitoring γ radiation emissions and determining appropriate isolation times for hyperthyroid cats treated with radioactive iodine (I(131)). Daily radiation emitted by 12 cats who had received a range of doses of I(131) (80-200 MBq) was measured and average background radiation readings deducted. The effective half-lives of the I(131) in the cats were found to have a median of 2.54 days (range 1.40-3.24 days). Cats treated with 200 MBq emitted 5 µGy/day more exposure than cats treated with lower doses throughout the study period (P=0.032). All cats were found to emit a total radiation dose exposure less than 100 µGy (range 0-43 µGy) during days 18-21 of isolation. The potential additional dose exposure to owners was calculated at various days that might be considered for the cats to be returned to their owners. Using this provisional data, maximum isolation periods at this institution could be safely reduced to 17 days as long as certain precautions are followed. This preliminary study demonstrated that this novel cage-side monitoring system can be used to calculate the effective half-life of I(131) and to measure γ radiation exposure from treated cats, which may assist other institutions in determining appropriate isolation times for individual cats.


Gender-specific associations between insulin resistance, hypertension, and markers of inflammation among adult Saudis with and without diabetes mellitus type 2.
In view of the important roles of the pro-inflammatory cytokines tumor necrosis factor alpha (TNF-α), interleukin-6 (IL-6) and the acute phase reactant C-reactive protein (CRP) in glucose metabolism and pathogenesis of diabetes mellitus type 2 (DMT2), we assessed gender-specific differences and relative associations of these inflammatory biomarkers to insulin resistance (IR) and risk markers for DMT2. Serum levels of TNF-α, IL-6 and CRP were determined in 119 clinically diagnosed DMT2 cases, 114 non-DMT2 subjects with IR, and 97 age-matched controls. Fasting blood samples were collected and serum glucose levels, lipid profile, and inflammatory markers were analyzed. In women, a significant association between elevated levels of IL-6 and risk of developing IR [Odds ratio (OR), 4.389, 95 % Confidence Interval (CI) 1.6-11.52, p = 0.004] was found. Significant associations were also found between elevated levels of CRP and risk of hypertension only in female subjects [OR (95% CI) 2.153 (1.04-4.53), p = 0.046]. While, in male subjects, a significant association between elevated levels of TNF-α and risk of developing IR [OR (95% CI) 2.32 (1.09-4.93), p = 0.029] was found. The present study showed apparent gender differences in the association of IL-6, CRP, and TNF-a with risks of IR and hypertension, and this could be attributed to sexual dimorphism in fat distribution.


Effect of human serum and some of its components on neutrophil adherence and migration across an epithelium.
The effect of human serum and some of its components on the process of transepithelial migration of human neutrophils was investigated in an in vitro system. 10% autologous serum caused an increase in neutrophil adherence to and migration across canine kidney epithelial cells. This increase in neutrophil binding also occurred if the epithelium but not the neutrophils had been preincubated with serum. The binding was lost if the serum was either preabsorbed over the kidney epithelium before use or heat inactivated. Indirect immunofluorescence studies indicated that IgG, IgM, and a component of C3 bound to the epithelial surface, whereas IgA, IgE, or C5a were not detectable. The majority of epithelial cells were immunofluorescent, however epithelial cells with varying degrees of reactivity were also apparent and approximately 5% of the epithelial cells did not bind IgG, IgM, and C3. When epithelia were simultaneously tested for the presence of either IgG, IgM, or C3, and bound neutrophils the few epithelial cells which did not bind IgG or IgM also did not bind C3 or neutrophils. Studies with monoclonal antibodies against Fc and C3 receptors indicate that neutrophil adherence to the epithelial surface was mediated predominately by the receptors for C3b and C3bi. In response to a chemotactic gradient, bound neutrophils were able to detach and migrate across the epithelium. A separate heat-stable factor(s) in serum was able to increase neutrophil migration across the epithelial monolayer. This factor acted independently of the factors which caused the increase in neutrophil binding as the increase in neutrophil migration also occurred under conditions (preabsorption over the kidney epithelium or heat inactivation) that prevented the increase in neutrophil binding. The increase in neutrophil migration may be caused by the permeability-increasing properties of this factor as both serum and heat-inactivated serum lowered the transepithelial electrical resistance an average of 38 and 35%, respectively, in 40 min. Upon removal of serum or heat-inactivated serum, the resistance returned 100 and 81%, respectively, in 5 h.


Obesity And Diabetes As Determinants Of Vitamin D Deficiency.
Vitamin D has been the focus of attention in the recent past owing to its multitude of effects on various organ systems including immune system, endocrine, cardiovascular etc. Diabetes mellitus and obesity are widely prevalent in our region. The present study was designed with an objective to determine the vitamin D status in relation to diabetes mellitus and obesity in our area. This cross-sectional study was conducted at the Medical C Unit of Department of Medicine, Ayub Teaching Hospital from January to December 2017. Approval of ethical committee was taken. A total of 117 patients were enrolled in this study of which 109 were finalized for analysis owing to incomplete data in 8 cases. Patients' characteristics were recorded on a structured proforma. Type 2 diabetes was confirmed using HbA1C Levels. Using ADA 2016 criteria. Vitamin D status was assessed using 25-OH-Vit D levels from the same laboratory. Height and weight of the patients were recorded to obtain BMI. Data was entered and analysed using SPSS version 20. Of the total sample, 69 (63.3%) were females and 40 (36.7%) were males. Mean age of the participants was 44.13±15.777. Mean vitamin D levels were 26.35±18.72. A total of 83 (76.14%) patients were either vitamin D deficient 66 (60.6%) or insufficient 17 (15.6%) while 26 (23.9%) were sufficient in vitamin D. There was statistically significant difference in vitamin D status in diabetic versus non diabetic patients (p=0.015). As regards BMI and vitamin D status, the difference was also statistically significant (p=0.018). Vitamin D deficiency is widely prevalent in our region. There is also a high prevalence of obesity and diabetes mellitus and they are inversely related to low vitamin D levels.


Biosafety Recommendations for Work with Influenza Viruses Containing a Hemagglutinin from the A/goose/Guangdong/1/96 Lineage.
The CDC and National Institutes of Health (NIH) Biosafety in Microbiological and Biomedical Laboratories (BMBL) manual describes biosafety recommendations for work involving highly pathogenic avian influenza (HPAI) (US Department of Health and Human Services [HHS], CDC. Biosafety in microbiological and biomedical laboratories, 5th ed. Atlanta, GA: CDC; 2009. HHS publication no. [CDC] 21-1112. Available at http://www.cdc.gov/biosafety/publications/bmbl5). The U.S. Department of Agriculture Guidelines for Avian Influenza Viruses builds on the BMBL manual and provides additional biosafety and biocontainment guidelines for laboratories working with HPAI (US Department of Agriculture, Animal and Plant Health Inspection Service, Agricultural Select Agent Program. Guidelines for avian influenza viruses. Washington, DC: US Department of Agriculture; 2011. Available at http://www.selectagents.gov/Guidelines_for_Avian_Influenza_Viruses.html). The recommendations in this report, which are intended for laboratories in the United States, outline the essential baseline biosafety measures for working with the subset of influenza viruses that contain a hemagglutinin (HA) from the HPAI influenza A/goose/Guangdong/1/96 lineage, including reassortant influenza viruses created in a laboratory setting. All H5N1 influenza virus clades known to infect humans to date have been derived from this lineage (WHO/OIE/FAO H5N1 Evolution Working Group. Continued evolution of highly pathogenic avian influenza A [H5N1]: updated nomenclature. Influenza Other Respir Viruses 2012;6:1-5). In 2009, the NIH Guidelines for Research Involving Recombinant or Synthetic Nucleic Acid Molecules were amended to include specific biosafety and biocontainment recommendations for laboratories working with Recombinant Risk Group 3 influenza viruses, including HPAI H5N1 influenza viruses within the Goose/Guangdong/1/96-like H5 lineage. In February 2013, the NIH guidelines were further revised to provide additional biosafety containment enhancements and practices for research with HPAI H5N1 viruses that are transmissible among mammals by respiratory droplets (i.e., mammalian-transmissible HPAI H5N1) (National Institutes of Health, Office of Biotechnology Activities. NIH guidelines for research involving recombinant or synthetic nucleic acid molecules. Appendix G-II-C-5: biosafety level 3 enhanced for research involving risk group 3 influenza viruses. Bethesda, MD: National Institutes of Health; 2013. Available at http://oba.od.nih.gov/rdna/nih_guidelines_oba.html). The recent revisions to the NIH guidelines focus on a smaller subset of viruses but are applicable and consistent with the recommendations in this report. The biosafety recommendations in this report were developed by CDC with advice from the Intragovernmental Select Agents and Toxins Technical Advisory Committee, which is a panel composed of federal government subject-matter experts, and from public input received in response to the request for information that was published in the Federal Register on October 17, 2012 (US Department of Health and Human Services, CDC. Influenza viruses containing the hemagglutinin from the Goose/ Guangdong/1/96 lineage; proposed rule; request for information and comment. 42 CFR, Part 73. Federal Register 2012;77:63783-5). Work with HPAI H5N1 virus should be conducted, at a minimum, at biosafety level 3 (BSL-3), with specific enhancements to protect workers, the public, animal health, and animal products. Original clinical specimens suspected of containing viruses of this lineage can only be handled at BSL-2 if the procedures do not involve the propagation of the virus. An appropriate biosafety level should be determined in accordance with a biosafety risk assessment. Additional information on performing biosafety risk assessments and establishing effective biosafety containment is available in the BMBL manual.


Novel CRYGC Mutation in Conserved Ultraviolet-Protective Tryptophan (p.Trp131Arg) Is Linked to Autosomal Dominant Congenital Cataract.
Congenital cataract (CC), the most prevalent cause of childhood blindness and amblyopia, necessitates prompt and precise genetic diagnosis. The objective of this study is to identify the underlying genetic cause in a Swiss patient with isolated CC. Whole exome sequencing (WES) and copy number variation (CNV) analysis were conducted for variant identification in a patient born with a total binocular CC without a family history of CC. Sanger Sequencing was used to confirm the variant and segregation analysis was used to screen the non-affected parents. The first de novo missense mutation at c.391T>C was identified in exon 3 of CRYGC on chromosome 2 causing the substitution of a highly conserved Tryptophan to an Arginine located at p.Trp131Arg. Previous studies exhibit significant changes in the tertiary structure of the crystallin family in the following variant locus, making CRYGC prone to aggregation aggravated by photodamage resulting in cataract. The variant can be classified as pathogenic according to the American College of Medical Genetics and Genomics (ACMG) criteria (PP3 + PM1 + PM2 + PS2; scoring 10 points). The identification of this novel variant expands the existing knowledge on the range of variants found in the CRYGC gene and contributes to a better comprehension of cataract heterogeneity.


Theories of schizophrenia: a genetic-inflammatory-vascular synthesis.
Schizophrenia, a relatively common psychiatric syndrome, affects virtually all brain functions yet has eluded explanation for more than 100 years. Whether by developmental and/or degenerative processes, abnormalities of neurons and their synaptic connections have been the recent focus of attention. However, our inability to fathom the pathophysiology of schizophrenia forces us to challenge our theoretical models and beliefs. A search for a more satisfying model to explain aspects of schizophrenia uncovers clues pointing to genetically mediated CNS microvascular inflammatory disease. A vascular component to a theory of schizophrenia posits that the physiologic abnormalities leading to illness involve disruption of the exquisitely precise regulation of the delivery of energy and oxygen required for normal brain function. The theory further proposes that abnormalities of CNS metabolism arise because genetically modulated inflammatory reactions damage the microvascular system of the brain in reaction to environmental agents, including infections, hypoxia, and physical trauma. Damage may accumulate with repeated exposure to triggering agents resulting in exacerbation and deterioration, or healing with their removal. There are clear examples of genetic polymorphisms in inflammatory regulators leading to exaggerated inflammatory responses. There is also ample evidence that inflammatory vascular disease of the brain can lead to psychosis, often waxing and waning, and exhibiting a fluctuating course, as seen in schizophrenia. Disturbances of CNS blood flow have repeatedly been observed in people with schizophrenia using old and new technologies. To account for the myriad of behavioral and other curious findings in schizophrenia such as minor physical anomalies, or reported decreased rates of rheumatoid arthritis and highly visible nail fold capillaries, we would have to evoke a process that is systemic such as the vascular and immune/inflammatory systems. A vascular-inflammatory theory of schizophrenia brings together environmental and genetic factors in a way that can explain the diversity of symptoms and outcomes observed. If these ideas are confirmed, they would lead in new directions for treatments or preventions by avoiding inducers of inflammation or by way of inflammatory modulating agents, thus preventing exaggerated inflammation and consequent triggering of a psychotic episode in genetically predisposed persons.


Qualitative and quantitative detection of SARS-CoV-2 RNA in untreated wastewater in Western Cape Province, South Africa.
Recent studies have shown that the detection of SARS-CoV-2 genetic material in wastewater may provide the basis for a surveillance system to track the environmental dissemination of this virus in communities. An effective wastewater-based epidemiology (WBE) system may prove critical in South Africa (SA), where health systems infrastructure, testing capacity, personal protective equipment and human resource capacity are constrained. In this proof-of-concept study, we investigated the potential of SARS-CoV-2 RNA surveillance in untreated wastewater as the basis for a system to monitor COVID-19 prevalence in the population, an early warning system for increased transmission, and a monitoring system to assess the effectiveness of interventions. The laboratory confirmed the presence (qualitative analysis) and determined the RNA copy number of SARS-CoV-2 viral RNA by reverse transcription polymerase chain reaction (quantitative) analysis from 24-hour composite samples collected on 18 June 2020 from five wastewater treatment plants in Western Cape Province, SA. The study has shown that a WBE system for monitoring the status and trends of COVID-19 mass infection in SA is viable, and its development and implementation may facilitate the rapid identification of hotspots for evidence-informed interventions.