Characterization of rifampicin-resistant Mycobacterium tuberculosis in Khyber Pakhtunkhwa, Pakistan.
Tuberculosis (TB), caused by Mycobacterium tuberculosis, is endemic in Pakistan. Resistance to both firstline rifampicin and isoniazid drugs (multidrug-resistant TB; MDR-TB) is hampering disease control. Rifampicin resistance is attributed to rpoB gene mutations, but rpoA and rpoC loci may also be involved. To characterise underlying rifampicin resistance mutations in the TB endemic province of Khyber Pakhtunkhwa, we sequenced 51 M. tuberculosis isolates collected between 2016 and 2019; predominantly, MDR-TB (n = 44; 86.3%) and lineage 3 (n = 30, 58.8%) strains. We found that known mutations in rpoB (e.g. S405L), katG (e.g. S315T), or inhA promoter loci explain the MDR-TB. There were 24 unique mutations in rpoA, rpoB, and rpoC genes, including four previously unreported. Five instances of within-host resistance diversity were observed, where two were a mixture of MDR-TB strains containing mutations in rpoB, katG, and the inhA promoter region, as well as compensatory mutations in rpoC. Heteroresistance was observed in two isolates with a single lineage. Such complexity may reflect the high transmission nature of the Khyber Pakhtunkhwa setting. Our study reinforces the need to apply sequencing approaches to capture the full-extent of MDR-TB genetic diversity, to understand transmission, and to inform TB control activities in the highly endemic setting of Pakistan.


Arnold's neuralgia caused by solitary fibrous tumor. Case report.
Solitary fibrous tumor (SFT) is a mesenchymal tumor which has been identified in a wide variety of localizations, including soft tissues, peritoneum, retroperitoneum, mediastinum, upper respiratory tract, nasopharyngeal sinuses, periosteum and extremities, orbit, major body cavities, intraspinal and intracranial localizations. The authors describe a case of SFT found in the neck of a young patient suffering from Arnold's neuralgia. After surgery, diagnosis of SFT was based on characteristic histopathological findings, especially on immunohistochemical positive staining for CD34 antigen. It has been described also the characteristic patterns making this diagnose sure, focusing the point that cure is possible with complete excision of the lesion.


Automatic acoustic classification of insect species based on directed acyclic graphs.
This work presents the design of a directed acyclic graph (DAG) scheme, the nodes of which incorporate hidden Markov models (HMMs) for classifying insect species. Such a DAG scheme is able to limit the problem space, while having the HMMs capture the temporal evolution of Mel-scaled spectrograms extracted out of wingbeat sounds. Interestingly, the proposed approach offers interpretability of the classification process by inspecting the sequence of edges activated in the DAG (path). The dataset encompasses 50 000 wingbeat sounds representing six species, i.e., Ae. aegypti (male and female), Cx. quinquefasciatus (male and female), Cx. stigmatosoma (male and female), Cx. tarsalis (male and female), Musca domestica, and Drosophila simulans, and is publicly available at https://sites.google.com/site/insectclassification/. Thorough species classification experiments showed that the proposed solution outperforms state-of-the-art approaches.


Survival on biventricular mechanical support with the Centrimag® as a bridge to decision: a single-center risk stratification.
Temporary mechanical assist devices are increasingly being used as a lifesaving bridge to decision in patients requiring cardiopulmonary resuscitation. We report our single-center experience with biventricular Centrimag® pumps over a five-year period. Data was retrospectively collected in consecutive patients who required biventricular support from 2008 to 2013. Patients who were supported with central cannulation using the Centrimag® system were analyzed. In addition to demographic information, data pertaining to indications, outcomes and mortality were collected. The cohort consisted of 48 patients (19 women and 29 men, mean age of 56 years). The median duration of support was 14 days. The median duration to patient expiration while still on the Centrimag® was 12 days. Thirty-day survival was 56% (27/48). Nine patients were explanted to recovery, while fourteen patients were converted to a durable LVAD, two of whom were then transplanted. We stratified patients into two groups. Group I comprised patients who were either explanted to recovery, converted to durable LVAD or transplanted (23/48) and Group II consisted of patients who either died on the Centrimag® or were explanted for withdrawal of care (25/48). Statistical analysis did not reveal any clinically significant differences between the two groups in terms of age, sex, etiology, hemodynamic, co-morbidities or laboratory parameters. The biventricular Centrimag® can be used as a bridge to decision in patients with thirty-day survival of >50%. Parameters to predict 30-day survival in this high-risk cohort continue to remain elusive.


[Serum alpha-1-fetoprotein and serum lactic dehydrogenase isoenzymes in liver tumours (author's transl)].
Serum alpha-1-fetoprotein (AFP) and serum lactic dehydrogenase isoenzymes (LDH I-V) were evaluated in healthy subjects as well as in 10 patients with primary liver carcinoma, in 10 patients with metastatic liver cancer and in 10 patients with cirrhosis of the liver. The diagnosis was established histologically in all cases. The upper limit of the normal AFP range was 9 ng/ml. Four out of all the patients with hepatocellular or cholangiocellular carcinomas had normal AFP values, 3 showed slightly increased AFP values, whilst a serum AFP concentration exceeding 174 ng/ml - limit which is statistically highly suggestive of hepatoma - was found in only 3 patients. Three out of the patients with metastatic liver cancer and 3 with cirrhosis showed moderately increased AFP values. In primary liver cancer LDH V is increased significantly and 8 out of all patients showed higher values of LDH V than LDH IV. By contrast, patients with metastatic liver cancer had significantly increased LDH IV, which was higher than LDH V in 9 out of all cases. Cirrhotics showed normal LDH isoenzymes. Combining these results, a definitive diagnosis could be made in 9 patients with primary carcinoma of the liver and in 9 patients with metastatic liver cancer.


BRAF inhibitor dabrafenib in patients with metastatic BRAF-mutant thyroid cancer.
Mutations of v-raf murine sarcoma viral oncogene homolog B (BRAF) are commonly identified in papillary and anaplastic thyroid carcinoma and are associated with worse prognosis compared with the wild type. BRAF inhibition in papillary thyroid carcinoma cell lines and xenografts inhibits proliferation and decreases downstream phosphorylation. Our objectives were to analyze safety and efficacy of the selective BRAF inhibitor dabrafenib in patients with metastatic BRAF-mutant thyroid carcinoma. We present the subset of patients with BRAF-mutant thyroid carcinoma enrolled in a larger phase 1 study, the main results of which are reported elsewhere. Fourteen patients with BRAF(V600E)-mutant thyroid carcinoma were enrolled, of whom 13 (93%) had received prior radioactive iodine. The median duration on treatment was 8.4 months, and seven (50%) patients received treatment for ≥10 months. The most common treatment-related adverse events were skin papillomas (n=8, 57%), hyperkeratosis (n=5, 36%), and alopecia (n=4, 29%), all of which were grade 1. Treatment-related adverse events grade ≥3 included grade 4 elevated lipase and grade 3 elevated amylase, fatigue, febrile neutropenia, and cutaneous squamous cell carcinoma (n=1 for each). Four (29%) partial responses were observed, and nine (64%) patients achieved at least 10% decrease. Only one responder progressed while on the study drug after a response duration of 9.3 months. The other three responders had not progressed, with response duration of 4.6+, 10.4+, and 21.4+ months. With seven (50%) patients showing no progression at the time of study completion, the median progression-free survival was 11.3 months. Dabrafenib was well tolerated and resulted in durable responses in BRAF-mutant differentiated thyroid carcinoma patients.


[A case of bilateral diaphragmatic paralysis clinically suggestive of brachial neuritis].
A 55-year-old man complained of acute onset of shoulder pain and dyspnea in the supine position. A diagnosis of bilateral diaphragmatic paralysis was made based on clinical inspection of his breathing pattern, radiographic appearance, and pulmonary function tests. He had had no traumatic or thoracic surgery or inflammatory episode. He did not suffer from diabetes, other central neural diseases, or any neoplastic disease. From the clinical feature and electromyographic findings, phrenic nerve involvement of brachial neuritis without any other muscle involvement was considered as a causative disease. When he received non-invasive intermittent positive-pressure ventilation by nasal mask in a supine position, his dyspnea was substantially attenuated and Carbon dioxide retention was lessened. After 1 year, his shoulder pain is still persisting and radiographic findings are not remarkably improved.


Effects of atrial natriuretic factor on pressor responsiveness to angiotensin II, norepinephrine, and vasopressin in conscious sheep.
The effects of pretreatment with atrial natriuretic factor (ANF) on the pressor responsiveness to injections of angiotensin II (ANGII), arginine vasopressin (AVP), and norepinephrine (NE), as well as the effect of pretreatment with ANGII on the hypotensive responses to ANF injection were studied in conscious sheep. The hemodynamic effects of ANF infusion (100 micrograms/h for 60 min) were also examined in animals pretreated with the angiotensin-converting enzyme (ACE) inhibitor, captopril. Infusion of ANF attenuated the pressor responsiveness to exogenous AII and NE, but caused no significant change in the blood pressure increases produced by vasopressin. In contrast, infusion of AII had no effect on the immediate hypotensive response to ANF injection. Infusion of ANF for 60 min produced similar hemodynamic actions in sheep during ACE inhibition as compared with the responses observed in normal sheep, although the reduction in cardiac output and increase in calculated total peripheral resistance was attenuated. Infusion of captopril increased plasma concentration of renin (PRC), and infusion of ANF produced no further change in PRC. In conclusion, the short-term cardiovascular responses to ANF infusion in conscious sheep are not mediated solely by inhibition of the renin-angiotensin system. However, ANF attenuates the pressor actions of pharmacologic doses of exogenous ANGII and NE. In contrast, the vasodepressor response to exogenous ANF injection was not altered in animals receiving ANGII infusion. This study suggests that ANF may be important in regulating the effects of endogenous vasoconstrictor hormones on blood pressure (BP).


Development and Testing of an Intelligent Pain Management System (IPMS) on Mobile Phones Through a Randomized Trial Among Chinese Cancer Patients: A New Approach in Cancer Pain Management.
Cancer has become increasingly prevalent in China over the past few decades. Among the factors that determine the quality of life of cancer patients, pain has commonly been recognized as a most critical one; it could also lead to the ineffective treatment of the cancer. Driven by the need for better pain management for cancer patients, our research team developed a mobile-based Intelligent Pain Management System (IPMS). Our objective was to design, develop, and test the IPMS to facilitate real-time pain recording and timely intervention among cancer patients with pain. The system's usability, feasibility, compliance, and satisfaction were also assessed. A sample of 46 patients with cancer pain symptoms were recruited at the Oncology Center of Xinhua Hospital affiliated to Shanghai Jiao Tong University School of Medicine, Chongming Branch (hereinafter referred to as "the Oncology Center"). In a pretest, participants completed a pain management knowledge questionnaire and were evaluated using the baseline cancer pain assessment and Karnofsky Performance Status (KPS) evaluation. The participants were then randomly assigned into two groups (the trial group and the control group). After a 14-day trial period, another round of cancer pain assessment, KPS evaluation and pain management knowledge assessment were repeated. In the trial group, the data were fully automatically collected by the IPMS. In the control group, the data were collected using conventional methods, such as phone interviews or door-to-door visits by physicians. The participants were also asked to complete a satisfaction questionnaire on the use of the IPMS. All participants successfully completed the trial. First, the feasibility of IPMS by observing the number of daily pain assessments recorded among patients was assessed. Second, the users' satisfaction, effectiveness of pain management, and changes in the quality of their lives were evaluated. All the participants gave high satisfaction score after they used IMPS. Both groups reported similar pain scores and KPS scores at the baseline. At the end of the trial, the mean pain score of the trial group was significantly lower than of the control group (P<.001). The ending KPS score of the trial group was significantly higher than of the control group (P<.001). The improvement of pain management knowledge score in the trial group was more pronounced than that in the control group (P<.001). This study provided preliminary data to support the potentials of using IPMS in cancer pain communication between patients and doctors and to provide real-time supportive intervention on a convenient basis at a low cost. Overall, the IPMS can serve as a reliable and effective approach to control cancer pain and improve quality of life for patients with cancer pain. Clinicaltrials.gov NCT02765269; http://clinicaltrials.gov/ct2/show/NCT02765269 (Archived by WebCite at http://www.webcitation.org/6rnwsgDgv).


[Analysis of the common respiratory viruses in children with acute respiratory infection in a hospital in Lanzhou City from 2021 to 2022].
To explore the situation of 8 common respiratory pathogens in children with acute respiratory infection (ARI) from 2021 to 2022.The retrospective study selected 8 710 ARI patients from September 2021 to August 2022 in the Maternal and Child Health Hospital of Gansu Province as the study object, patients aged 0 to 17 years old, including 5 048 male children and 3 662 female children. Indirect immunofluorescence was used to detect 8 common respiratory pathogens, including influenza virus A (FluA), influenza virus B (FluB), parainfluenza virus (PIV), respiratory syncytial virus (RSV), adenovirus (ADV), Mycoplasma pneumoniae (MP), Chlamydia pneumoniae (CP), and Coxsackie virus group B (CoxB) IgM antibodies. χ2 test was used to analyze the results. The results showed that 1 497 of 8 710 children with ARI were positive, with a positive rate of 17.19%. The detection rate of MP among 8 common respiratory pathogens was 11.34%, accounting for 66.0%, followed by FluB, CoxB, PIV, RSV, ADV, FluA and CP, accounting for 13.83%, 9.55%, 6.01%, 2.61%, 1.47%, 0.40% and 0.13%, respectively. Respiratory tract viruses (FluA, FluB, RSV, ADV, PIV, CoxB) accounted for 33.86%.There were significant differences in the detection rates of PIV, ADV and MP among children of different genders (χ2=6.814, 5.154 and 17.784, P<0.05). The detection rate of school-age children (6-17 years old) was the highest, accounting for 33.27% (184/553). The detection rates of 8 common respiratory pathogens in patients with ARI were higher in spring and winter and lower in summer and autumn. To sum up, from 2021 to 2022, MP and FluB infection were dominant in ARI patients in our hospital. The peak period of 8 common respiratory pathogens was in spring and winter. The physical examination rate of 8 common respiratory pathogens in ARI patients aged 6-17 years old was the highest.


CTRP1 Attenuates Cerebral Ischemia/Reperfusion Injury via the PERK Signaling Pathway.
Cerebral ischemic stroke is one of the leading causes of death worldwide. Previous studies have shown that circulating levels of CTRP1 are upregulated in patients with acute ischemic stroke. However, the function of CTRP1 in neurons remains unclear. The purpose of this study was to explore the role of CTRP1 in cerebral ischemia reperfusion injury (CIRI) and to elucidate the underlying mechanism. Middle cerebral artery occlusion/reperfusion (MCAO/R) and oxygen-glucose deprivation/reoxygenation (OGD/R) models were used to simulate cerebral ischemic stroke in vivo and in vitro, respectively. CTRP1 overexpression lentivirus and CTRP1 siRNA were used to observe the effect of CTRP1 expression, and the PERK selective activator CCT020312 was used to activate the PERK signaling pathway. We found the decreased expression of CTRP1 in the cortex of MCAO/R-treated rats and OGD/R-treated primary cortical neurons. CTRP1 overexpression attenuated CIRI, accompanied by the reduction of apoptosis and suppression of the PERK signaling pathway. Interference with CTRP1 expression in vitro aggravated apoptotic activity and increased the expression of proteins involved in the PERK signaling pathway. Moreover, activating the PERK signaling pathway abolished the protective effects of CTRP1 on neuron injury induced by CIRI in vivo and in vitro. In conclusion, CTRP1 protects against CIRI by reducing apoptosis and endoplasmic reticulum stress (ERS) through inhibiting the PERK-dependent signaling pathway, suggesting that CTRP1 plays a crucial role in the pathogenesis of CIRI.


The topography of the furcation entrance in Chinese molars. Furcation entrance dimensions.
The purpose of the present study was to document the furcation entrance dimensions (FEDs) of the maxillary and mandibular 1st and 2nd molars and relate them to the choice of periodontal therapy. Study samples consisted of 89 maxillary molars (49 1st and 40 2nd molars) and 93 mandibular molars (50 1st and 43 2nd molars). All the FEDs of the molars were examined and measured under a stereomicroscope at 2.5 x equipped with a Bioscan OPTIMAS Image Analyzer (BOIA). The results may be summarized as follows. (1) The mean FEDs in the buccal, distal and mesial furcations of maxillary 1st and 2nd molars were 0.74 mm, 0.99 mm and 1.04 mm in the 1st molars, and 0.63 mm, 0.67 mm, 0.90 mm in the 2nd molars, respectively. In the buccal and lingual furcations of mandibular 1st and 2nd molars, they measured 0.88 mm and 0.81 mm, and 0.73 mm and 0.71 mm, respectively. (2) The %s of FEDs of 0.56 mm or less (the tip width of a Cavitron tip being 0.56 mm) in the buccal, distal and mesial furcations of maxillary 1st and 2nd molars, accounted for 32%, 8% and 6% of 1st molars, and 40%, 40% and 18% of 2nd molars. In the buccal and lingual areas of mandibular 1st and 2nd molars, they accounted for 16% and 26%, and 35% and 33% of the furcations, respectively.(ABSTRACT TRUNCATED AT 250 WORDS)


Variation in the use of point-of-care ultrasound in general practice in various European countries. Results of a survey among experts.
Before implementing point-of-care ultrasound in general practice in Denmark, we sought inspiration from other countries/regions. To collect information about the use and organizational aspects of point-of-care ultrasound in general practice in different European countries/regions. Fifteen key persons with knowledge about the use of ultrasound in general practice in Austria, Catalonia, Denmark, Finland, Germany, Greenland, Iceland, the Netherlands, Norway, Scotland, Sweden, and Switzerland were included. Participants received a link to a web-based questionnaire. The primary outcome measures were educational aspects regarding the use of point-of-care ultrasound; clinical application and use; financial aspects; and main barriers to using ultrasound in general practice. In eight out of 12 countries/regions there were national societies for the use of ultrasound in general practice. The respondents from three countries/regions reported that the use of ultrasound was integrated into undergraduate medical education. In nine of the countries/regions, there was formalized training for general practitioners, but only three reported this to be part of the specialization to become a general practitioner. In seven out of 12 countries/regions, general practitioners received payment for ultrasound scans. However, the payment and the requirements for reimbursement differed between countries. Lack of time, lack of training, and financial aspects were important common barriers across countries/regions. There were significant differences regarding the use and organizational aspects of point-of-care ultrasound in general practice in Europe. Lack of time and training as well as financial aspects were important barriers to the use of point-of-care ultrasound in general practice.[Box: see text].


The calcium-binding protein VILIP in olfactory neurons: regulation of second messenger signaling.
Visinin-like-protein (VILIP), a member of the neuronal subfamily of EF-hand calcium-sensor proteins is shown to be expressed in olfactory sensory cells of the rat nasal epithelium. Its prominent localization in cilia and dendritic knobs-the chemosensory compartments of olfactory neurons-suggests that the calcium-binding protein could be involved in olfactory signal transduction. Consistent with this assumption, it was found that recombinant VILIP attenuates in a calcium-dependent manner odorant-induced cAMP formation in olfactory cilia preparations. Kinetic data indicate that VILIP does not interfere with odorant-induced receptor desensitization. Since VILIP inhibits the forskolin-induced formation of cAMP, it is conceivable that VILIP may directly affect the olfactory adenylyl cyclase. Thus, VILIP may play a role in adaptation of olfactory neurons.


Short- and long-term therapy with tocainide for malignant ventricular tachyarrhythmias.
Tocainide was administered to 228 patients referred for treatment of recurrent ventricular tachyarrhythmias that were refractory to therapy with conventional antiarrhythmic drugs. After baseline studies, 1200 to 2400 mg tocainide/day was given for 4 days. Tocainide was effective in 49% of 180 patients evaluated with monitoring and exercise testing and in 35% of 48 patients undergoing electrophysiologic testing. No clinical parameter predicted the response to tocainide, although there was a correlation with the effect of lidocaine. Tocainide was selected for long-term treatment in 73 patients who were followed for an average of 26.4 months (range 1 to 92 months). The incidence of sudden death was 4.3% per year and two patients had nonfatal recurrence of arrhythmia. It is concluded that tocainide is effective and well tolerated during long-term use if therapy is evaluated carefully and is individualized.


[Ultrasonic sludge treatment and its application on aerobic digestion].
In order to enhance the degradation efficiency of waste activated sludge (WAS) in conventional aerobic digestion, various ultrasonic assisted treatment methods were investigated including ultrasonic disintegration of influent sludge, ultrasonic improvement of influent sludge activity and ultrasonic disintegration of return sludge. Firstly the effects of ultrasonic sludge treatment were studied to choose appropriate ultrasonic parameters, and then the experiments of aerobic digestion with different ultrasonic treatments were carried out. The results show that 1.0 W/mL, 10 minutes ultrasonic treatment can increase soluble chemical oxygen demand (SCOD) in the supernatant phase of sludge sample by 5.4 times and decrease total suspended solid (TSS) by 16%; 0.05 W/mL, 10 min ultrasonic treatment can increase the specific oxygen uptake rate (SOUR) of sludge sample by 29%. The two kinds of ultrasonic influent sludge pretreatment can't improve aerobic digestion effectively. Ultrasonic return sludge disintegration can enhance the volatile suspended solid (VSS) degradation ratio by 15%. Furthermore, the settlement performance of digested sludge is still good and the pollutant concentrations of supernatant phase increase slightly. So ultrasonic return sludge disintegration is considered as the most appropriate assisted treatment mode for aerobic digestion.


Pathogenicity and phenotypic sulfadiazine resistance of Toxoplasma gondii isolates obtained from livestock in northeastern Brazil.
Toxoplasma gondii is the causative protozoan agent of toxoplasmosis, which is a common infection that is widely distributed worldwide. Studies revealed stronger clonal strains in North America and Europe and genetic diversity in South American strains. Our study aimed to differentiate the pathogenicity and sulfadiazine resistance of three T. gondii isolates obtained from livestock intended for human consumption. The cytopathic effects of the T. gondii isolates were evaluated. The pathogenicity was determined by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) using a CS3 marker and in a rodent model in vivo. Phenotypic sulfadiazine resistance was measured using a kinetic curve of drug activity in Swiss mice. IgM and IgG were measured by ELISA, and the dihydropteroate synthase (DHPS) gene sequence was analysed. The cytopathic effects and the PCR-RFLP profiles from chickens indicated a different infection source. The Ck3 isolate displayed more cytopathic effects in vitro than the Ck2 and ME49 strains. Additionally, the Ck2 isolate induced a differential humoral immune response compared to ME49. The Ck3 and Pg1 isolates, but not the Ck2 isolate, showed sulfadiazine resistance in the sensitivity assay. We did not find any DHPS gene polymorphisms in the mouse samples. These atypical pathogenicity and sulfadiazine resistance profiles were not previously reported and served as a warning to local health authorities.


Promoting continuity of care for homeless adults with unmet health needs: The role of brief interventions.
Promoting timely and continuous care for people experiencing homelessness has been a challenge in many jurisdictions, plagued by access barriers and service fragmentation. As part of a larger programme evaluation, this study used qualitative methods to examine the role of a brief interdisciplinary intervention in supporting continuity of care for this population in a large Canadian urban centre. The intervention provides time-limited case management, primary and psychiatric care, and peer accompaniment to homeless adults with unmet health needs discharged from hospital. Data were collected from 52 study participants between July 2013 and December 2014. Three focus groups were conducted with service providers and people with lived experience of homelessness, and 29 individual, semi-structured interviews were conducted with service users and other key informants. Transcripts were analysed using thematic analysis. Analysis was informed by existing frameworks for continuity of care, while remaining open to additional or unexpected findings. Findings suggest that brief interdisciplinary interventions can promote continuity of care by offering low-barrier access, timely and responsive service provision, including timely connection to long-term services and supports, appropriate individualised services and effective co-ordination of services. Although brief interdisciplinary interventions were perceived to promote access, timeliness and co-ordination of care for this population with complex health and social needs, gaps in the local service delivery context can present persisting barriers to care comprehensiveness and continuity.


The epileptic pre-aura.
The term "pre-aura" is used for the designation of earliest pre-ictal manifestations that escape detection by conventional EEG and both objective and subjective behavioral signs. Fairly recent reports have indicated earliest pre-ictal vascular changes (perifocal ischemia and hyperemia) and also changes found in various methods of quantified EEG. These changes may precede the seizure onset (in conventional terms) by 1 to 20 minutes. There is good reason to assume that pre-auras are limited to focal epilepsies and do not occur in the various subforms of primary generalized epilepsy. The electrodecremental type of seizure onset, mostly found in the frontal region, could also denote a pre-aura in the earliest decremental phase. The very earliest pre-aura phenomena are most likely to be neuronal rather than vascular. Further proof could come with the use of ultrafast EEG frequency recordings.


Pulmonary amyloidosis and multiple myeloma mimicking lymphoma in a patient with Sjogren's syndrome: A case report.
Sjogren's syndrome (SS), which affect salivary gland function, is an autoimmune disease. SS may involve extraglandular organs. Approximately 10 to 20 percent of SS patients have clinically significant lung disease, but presentation of pulmonary amylodosis is extremly rare. The incidence of benign monoclonal gammopathy in SS patients is high, but multiple myeloma is rare. No case involving the simultaneous occurrence of two rare diseases, pulmonary amyloidosis and multiple myeloma, in the same patient with SS has been reported so far. A 41-year-old male patient was referred to our hematology department due to incidentally detected gastric plasmacytoma. He had been diagnosed with SS four years earlier. Multiple miliary nodules, ground glass opacity in both lung fields, and enlargement of both inguinal lymph nodes was observed on chest and abdomen computer tomography. Based on the pathological findings of lung and lymph node biopsied specimens, the patient was diagnosed with pulmonary amyloidosis and multiple myeloma. Pulmonary amyloidosis and multiple myeloma associated with SS has rarely been reported. This is an extremely rare case of simultaneous pulmonary amyloidosis and multiple myeloma in the same patient with SS.


Integrin connections map: to infinity and beyond.
Integrins are transmembrane proteins that serve as primary sensors of the extracellular matrix (ECM) environment. In response to interactions with the ECM, integrins initiate signaling pathways that regulate cell migration, growth, and survival. Advances in imaging have contributed to the understanding of the dynamic nature of these cell-ECM interactions and the complexes that form at these sites and have provided insights into their regulation and signal organizing functions.


The role of microRNAs in neurons and neuroimmune communication in the dorsal root ganglia in chronic pain.
Despite being a life-restricting condition, chronic pain remains poorly treated. A better understanding of the underlying mechanisms of chronic pain and thence development of innovative targets is therefore essential. Recently we have started to elucidate the importance of the role of microRNAs (miRs) in preclinical chronic pain. miRs are small, non-coding RNAs that regulate genes including those involved in nociceptive signalling. MiRs can exert their effects both intracellularly and extracellularly, the latter of which requires that they are released either as naked species or packaged in exosomes. Here we discuss changes in miR expression that occur in the dorsal root ganglia in murine models of chronic pain. We consider the downstream targets of changes in miR expression, including voltage-gated ion channels, as well as discuss extracellular consequences such as changes in macrophage phenotype that constitute of means by which neuron-immune cell crosstalk occurs. Such miR-mediated intracellular communication could provide a novel target for the treatment of chronic pain, which would be most effective if tailored to the specific cause of pain. Indeed, we conclude by reviewing evidence for the involvement of miRs in clinical cases of chronic pain, supporting the notion that tailored, miR-targeted therapies could prove to be an effective new strategy for the treatment of chronic pain clinically.


The Euphausia superba transcriptome database, SuperbaSE: An online, open resource for researchers.
Antarctic krill (Euphausia superba) is a crucial component of the Southern Ocean ecosystem, acting as the major link between primary production and higher trophic levels with an annual predator demand of up to 470 million tonnes. It also acts as an ecosystem engineer, affecting carbon sequestration and recycling iron and nitrogen, and has increasing importance as a commercial product in the aquaculture and health industries. Here we describe the creation of a de novo assembled head transcriptome for E. superba. As an example of its potential as a molecular resource, we relate its exploitation in identifying and characterizing numerous genes related to the circadian clock in E. superba, including the major components of the central feedback loop. We have made the transcriptome openly accessible for a wider audience of ecologists, molecular biologists, evolutionary geneticists, and others in a user-friendly format at SuperbaSE, hosted at http://www.krill.le.ac.uk.


Three-dimensional total synchronous luminescence spectroscopy criteria for discrimination between normal and malignant breast tissues.
Specimens of malignant and normal female human breast tissues were analyzed after surgery by means of synchronous luminescence spectroscopy. Measurements were performed in the ranges of excitation wavelengths from 330 to 650 nm and synchronous wavelengths from 30 to 120 nm to obtain ordinary and first derivative three-dimensional total synchronous luminescence spectra (3d-TSLS) of each specimen. Arithmetic mean of these spectra has been calculated for normal and malignant specimens and analyzed to establish criteria for tissue differentiation. Spectral domain volumes (volumes below luminescence intensity surface) and mean spectral slopes have been calculated and also analyzed as tissue discrimination criteria. The obtained results are discussed in view of the possible relevance of synchronous luminescence spectroscopy in discrimination between normal and malignant breast tissue.


Impact of telephonic interviews on persistence and daily adherence to insulin treatment in insulin-naïve type 2 diabetes patients: dropout study.
The objective of this study is to evaluate the impact of sequential telephonic interviews on treatment persistence and daily adherence to insulin injections among insulin-naïve type 2 diabetes patients initiated on different insulin regimens in a 3-month period. A total of 1,456 insulin-naïve patients with type 2 diabetes (mean [standard deviation, SD] age: 56.0 [12.0] years, 49.1% were females) initiated on insulin therapy and consecutively randomized to sequential (n=733) and single (n=723) telephonic interview groups were included. Data on insulin treatment and self-reported blood glucose values were obtained via telephone interview. Logistic regression analysis was performed for factors predicting increased likelihood of persistence and skipping an injection. Overall, 76.8% patients (83.2% in sequential vs 70.3% in single interview group, (P<0.001) remained on insulin treatment at the third month. Significantly higher rate for skipping doses was noted in basal bolus than in other regimens (27.0% vs 15.0% for premixed and 15.8% basal insulin, respectively, P<0.0001). Logistic regression analysis revealed sequential telephonic interview (odds ratio [OR], 1.531; 95% confidence interval [CI], 1.093-2.143; P=0.013), higher hemoglobin A1c levels (OR, 1.090; 95% CI, 0.999-1.189; P=0.049), and less negative appraisal of insulin therapy as significant predictors of higher persistence. Basal bolus regimen (OR, 1.583; 95% CI, 1.011-2.479; P=0.045) and higher hemoglobin A1c levels (OR, 1.114; 95% CI, 1.028-1.207; P=0.008) were the significant predictors of increased likelihood of skipping an injection. Our findings revealed positive influence of sequential telephonic interview, although including no intervention in treatment, on achieving better treatment persistence in type 2 diabetes patients initiating insulin.


Targeting the pericyte antigen DLK1 with an alpha type-1 polarized dendritic cell vaccine results in tumor vascular modulation and protection against colon cancer progression.
Despite the availability of various treatment options, colorectal cancer (CRC) remains a significant contributor to cancer-related mortality. Current standard-of-care interventions, including surgery, chemotherapy, and targeted agents like immune checkpoint blockade and anti-angiogenic therapies, have improved short-term patient outcomes depending on disease stage, but survival rates with metastasis remain low. A promising strategy to enhance the clinical experience with CRC involves the use of dendritic cell (DC) vaccines that incite immunity against tumor-derived blood vessels, which are necessary for CRC growth and progression. In this report, we target tumor-derived pericytes expressing DLK1 with a clinically-relevant alpha type-1 polarized DC vaccine (αDC1) in a syngeneic mouse model of colorectal cancer. Our pre-clinical data demonstrate the αDC1 vaccine's ability to induce anti-tumor effects by facilitating cytotoxic T lymphocyte activity and ablating the tumor vasculature. This work, overall, provides a foundation to further interrogate immune-mediated mechanisms of protection in order to help devise efficacious αDC1-based strategies for patients with CRC.


Assessment of the effect of alpha-galactosides injected during embryogenesis on selected chicken traits.
The effect of different doses of alpha-galactoside (RFOs) preparations from Pisum sativum L. cv. Opal, injected into eggs during embryogenesis, on maintaining a high number of bifidobacteria, selected chicken broiler traits and the lipoprotein level of blood were studied. Two independent experiments were conducted. In the first, Ringer water solution containing 1.763 mg/egg of fructooligosaccharides (FOS) (I group), 2.1158 mg of pea RFO preparation containing 20% sucrose (II group) and 0.4232 mg of sucrose (III group) were injected into Hubbard broiler breeder eggs containing 12-day old embryos. Only Ringer water solution was applied to the eggs of the control group (IV group). The number of bifidobacteria determined in faeces of two-day old chicken of groups I and II was significantly higher in comparison with the sucrose and control groups. The high level ofbifidobacteria of groups I and II was maintained during 6 weeks. The dose of both preparations had no influence on the body weight, carcass, breast muscle, leg and abdominal fat ratio, total cholesterol, HDL and LDL serum concentrations. Broiler mortality and breast muscle cholesterol concentration was highest (P < 0.05) for the control group. On the other hand, the European Production Index, as well as serum triglycerides, were the lowest for this group. The second experiment was performed on Hybro G chicken breeder eggs. 0.69, 3.43 and 6.87 mg/egg of pea RFO preparation doses containing 20% sucrose were injected into the experimental groups. The number of bifidobacteria in the caecum and selected meat traits of broilers were determined. The results of this experiment confirmed that RFO injection in ovo causes the long-time maintenance of a high level ofbifidobacteria. The dose of the preparations does not have any effect on the selected broiler meat traits, except that the highest dose increases the percent of carcase in body weight. However, this dose reduced the hatchability of the treated embryos.


Establishing a Cutoff Serum Thyroglobulin Value for the Diagnosis and Management of Well-Differentiated Thyroid Cancer.
Objective The aim of this study was to define a cutoff serum thyroglobulin (Tg) level associated with either residual or metastasis that may help decide postoperative radioactive iodine (RAI) scan and treatment in differentiated thyroid cancer (DTC) patients residing in low-income countries like Nepal. Methods We prospectively studied a total of 81 patients (female-to-male ratio of 3.0:1; mean age: 37.3 ± 14.0 years, within age range of 14-88 years) who underwent total thyroidectomy with/without neck dissection and were referred for RAI whole-body scan (WBS) ± RAI ablation or adjuvant treatment in the department of Nuclear Medicine, Chitwan Medical College. We calculated the cutoff value of Tg using receiver operating characteristic (ROC) curve analysis. Results Forty-six of 81 patients (56.7%) had remnants in the thyroid bed, 26/81 (32.1%) had regional lymph node metastasis, 9/81 (11.1%) had distant lymph node metastasis, 3/81 (3.7%) had lung metastases, and only 1/81 (1.2%) had bone metastases. RAI WBS was positive in 61/81 (75.3%) patients and negative in 20/81 (24.7%) patients. Seventeen of 81 (20.9%) patients had negative RAI scans with low serum Tg levels; only 3/81 (3.7%) patients had Tg elevated negative RAI scan (TENIS). Although scan was positive in 61/81 (75.3%) patients, 64/81 (79.0%) patients received treatment with RAI, of which 3/81 (3.7%) patients were TENIS patients. There was a significant difference in serum Tg levels between patients who received or did not receive RAI ablation or treatment ( p < 0.05). On ROC curve analysis, the cutoff value of Tg levels between patients who received and did not receive treatment was 2.9 ng/mL (sensitivity: 85.9%; specificity: 94.1%; positive predictive value [PPV], 98.2%; negative predictive value [NPV]: 64.0%; AUC: 0.938). Conclusion We identified a cutoff value of 2.9 ng/mL between patients who required or did not require treatment with high sensitivity, specificity, and PPVs.


To get hold of the wrong end of the stick: reasons for poor idiom understanding in children with reading comprehension difficulties.
The aim was to identify the source of idiom understanding difficulties in children with specific reading comprehension failure. Two groups (ns = 15) of 9- to 10-year-olds participated. One group had age-appropriate word reading and reading comprehension; the other group had age-appropriate word reading but poor reading comprehension. Each child completed an independent assessment of semantic analysis skills and 2 multiple-choice assessments of idiom comprehension. In 1 assessment, idiomatic phrases were embedded in supportive story contexts; in the other assessment, they were presented out of context. Performance on transparent idioms (which are amenable to interpretation by semantic analysis) and opaque idioms (which can only be interpreted by inference from context if the meaning is not known) was compared. The groups demonstrated comparable semantic analysis skills and understanding of transparent idioms. Children with poor comprehension were impaired in the use of supportive context to aid their understanding of the opaque idioms. The study identifies poor inference from context as a source of the idiom understanding difficulties in children with poor reading comprehension; there was no evidence that poor semantic analysis skills contributed to their difficulties. Children with poor comprehension should be supported in the use of context to understand unfamiliar figurative language.


Overcoming multidrug resistance of cancer cells by direct intranuclear drug delivery using TAT-conjugated mesoporous silica nanoparticles.
The development of multidrug resistance (MDR) in cancer cells is one of major obstacles to the effective cancer chemotherapy. In this report we demonstrate the effective circumvention of multidrug resistance in cancer cells by an active nuclear-targeted drug delivery system that was constructed by conjugating TAT peptide onto the surface of mesoporous silica nanoparticles (MSNs-TAT). The conjugation of TAT peptide facilitated the intranuclear localization of MSNs-TAT and the release of the encapsulated drugs directly within the nucleoplasm. The direct intranuclear drug delivery of doxorubicin (DOX) in multidrug resistant MCF-7/ADR cancer cells was capable of increasing the intracellular as well as intranuclear drug concentrations much more effectively than free DOX or delivered by MSNs in the absence of TAT peptide. With the nuclear drug delivery fashion, DOX-MSNs-TAT presents a promising strategy in overcoming MDR in cancer cells and improving the therapeutic index of currently available chemotherapeutics by enhancing therapeutic efficacy and reducing side effects.


Compatible solutes and fungal development.
Compatible solutes are components that can be quickly accumulated and degraded inside fungal cells. They do not disturb the functioning of proteins and protect the cell under adverse conditions. In this issue of the Biochemical Journal, Solomon and co-workers evaluate the role of mannitol, one of these components, in Stagonospora nodorum, a plant-pathogenic fungus, and find surprising effects on the development of spores and spore-forming structures.


Comparison of finger and forehead oximetry sensors in postanesthesia care patients.
We compared loss of pulse oximetry signal (dropout rates) for both finger and forehead sensors in postanesthesia patients. Pulse oximetry is a widely practiced method for measuring oxygen saturation. Several studies in various patient populations have demonstrated that low flow states, patient movement, and hypothermia may result in poor signal quality with the use of finger oximetry sensors. These clinical conditions are common in patients as they emerge from anesthesia. New forehead sensors may reduce signal dropout. A method-comparison design was used to compare finger and forehead oximetry signal dropout rates. Of 48 subjects studied, only three had a signal dropout. Overall, there were seven episodes of signal dropout; six of seven occurred with the finger sensor. Signal dropout occurred rarely in PACU subjects. Use of finger sensors for routine postanesthesia monitoring should be adequate in the majority of patients.


Metastatic squamous cell carcinoma to the pancreas: Report of an extremely rare case.
Secondary pancreatic lesions are very uncommon. Resection of these metastatic lesions with a curative intent has been reported in selected patients; however, the survival benefit from these procedures has yet to be clearly determined. A 78-year-old male patient presented to our department with obstructive jaundice. Three years prior to presentation the patient had undergone right pneumonectomy, due to stage IIA, low-grade squamous cell lung carcinoma. After resection, the patient received adjuvant chemotherapy, with no evidence of local or systemic recurrence over the following 3 years. Abdominal computer tomography on admission revealed a 3-cm lesion located at the head of the pancreas, causing biliary obstruction. Endoscopic ultrasound biopsy revealed malignant cells, suspicious for squamous cell carcinoma. Due to the limited extent of the metastatic disease and in view of the patient's good condition, surgical resection was proposed and the patient successfully underwent pancreaticoduodenectomy. Histological examination of the resected specimen confirmed a squamous cell carcinoma, with an immunochemical profile similar to that of the primary lung tumor. Therefore, pancreatic resection with curative intent may be feasible in selected patients with secondary metastatic tumors. However, further studies are required in order to determine the benefit of these major procedures in terms of survival outcomes.


TRPC channels determine human keratinocyte differentiation: new insight into basal cell carcinoma.
Aberrant keratinocyte differentiation is considered to be a key mechanism in the onset of hyperproliferative dermatological diseases, including basal cell carcinoma (BCC). It is, therefore, vital to understand what drives keratinocytes to develop such pathological phenotypes. The role of calcium in keratinocyte differentiation is uncontested but the mechanisms controlling calcium-induced differentiation have yet to be completely elucidated. This study was designed to investigate the role of calcium-permeable TRPC channels in human keratinocyte differentiation and BCC, using a combination of molecular and cell biology approaches, involving electrophysiology and Ca(2+)-imaging, on the HaCaT cell line, primary cultures of normal human keratinocytes, and BCC cells. We demonstrated that TRPC1/TRPC4 channel expression was important for keratinocyte differentiation, as knocking out these channels (by siRNA strategy) prevented the induction of Ca(2+)-induced differentiation. TRPC1/TRPC4-mediated calcium entry and endoplasmic reticulum Ca(2+) content increased significantly in differentiated keratinocytes. However, the failure of BCC cells to differentiate was related to a lack of TRPC channel expression and calcium entry. In summary, our data demonstrate that TRPC1 and TRPC4 channels are key elements in keratinocyte Ca(2+) homeostasis and differentiation and may therefore be responsible for skin pathologies.


MicroRNA expression profiles associated with mutational status and survival in malignant melanoma.
Malignant cutaneous melanoma is a highly aggressive form of skin cancer. Despite improvements in early melanoma diagnosis, the 5-year survival rate remains low in advanced disease. Therefore, novel biomarkers are urgently needed to devise new means of detection and treatment. In this study, we aimed to improve our understanding of microRNA (miRNA) deregulation in melanoma development and their impact on patient survival. Global miRNA expression profiles of a set of melanoma lymph node metastases, melanoma cell lines, and melanocyte cultures were determined using Agilent array. Deregulated miRNAs were evaluated in relation with clinical characteristics, patient survival, and mutational status for BRAF and NRAS. Several miRNAs were differentially expressed between melanocytes and melanomas as well as melanoma cell lines. In melanomas, miR-193a, miR-338, and miR-565 were underexpressed in cases with a BRAF mutation. Furthermore, low expression of miR-191 and high expression of miR-193b were associated with poor melanoma-specific survival. In conclusion, our findings show miRNA dysregulation in malignant melanoma and its relation to established molecular backgrounds of BRAF and NRAS oncogenic mutations. The identification of an miRNA classifier for poor survival may lead to the development of miRNA detection as a complementary prognostic tool in clinical practice.


Therapeutic strategy and outcome of spine tumors in pregnancy: a report of 21 cases and literature review.
A retrospective study was performed. To illustrate the characteristics of spine tumors during pregnancy and obtain better insight into therapeutic strategies of such tumors by analyzing 21 cases treated in Changzheng Hospital and reviewing previous reports in the literature. The concurrence of spine tumors and pregnancy is relatively rare. There are controversies over the treatment options for this disease, which increase the difficulty of the clinical treatment. Between 2002 and 2013, 21 pregnant patients were identified with spine tumors. Clinical data including symptoms, signs, treatment options, and obstetrical and neonatal outcomes were recorded and preserved. Clinical data and treatment efficacy were analyzed via medical record review. The median age of the 21 patients was 28.87 years (interquartile range, 6.00 yr). Tumor types in this series were giant cell tumor (5 cases), hemangioma (5 cases), schwannoma (4 cases), eosinophilic granuloma (2 cases), neurofibroma (1 case), multiple myeloma (1 case), and with metastatic tumor (3 cases). Two patients underwent spine surgery during pregnancy and 8 patients accepted tumor resection immediately after delivery. Pregnancy termination occurred in 5 patients, whereas the rest of the patients smoothly gave birth to healthy babies including 3 premature infants. Two patients died and 2 patients experienced local recurrence during follow-up. With close observation, it was found that most of pregnant patients with benign spine tumors could postpone surgery after delivery. Surgical treatment should be adopted during pregnancy when patients are with highly malignant tumor or experience a sharp deterioration and the guard of it is safer than radiotherapy and chemotherapy during pregnancy. 4.


A case of desmoplastic ameloblastoma of the maxilla.
A case of desmoplastic ameloblastoma of the maxilla is presented. The tumor showed characteristic radiographical and histological features. Radiographically, the tumor showed a mixed radiolucent-radiopaque lesion occupying the left maxillary sinus with undefined borders in the alveolar process. Histopathologically, the tumor was characterized by abundant, densely collagenous stroma with small nests and strands of odontogenic epithelium and bone tissues. Findings during operation suggested that the tumor arising in the alveolar process had grown upwards and almost displaced the left maxillary sinus. Complete resection is recommended for this variant because of the possibility of tumor infiltration into the surrounding bone marrow spaces.


MicroRNA-30b Regulates High Phosphorus Level-Induced Autophagy in Vascular Smooth Muscle Cells by Targeting BECN1.
Autophagy is an evolutionarily conserved mechanism that affects the survival and functions of vascular smooth muscle cells (VSMCs). We explored the role of microRNAs (miRNAs) in regulating autophagy in VSMCs exposed to high phosphorus (Pi) levels. VSMCs were isolated from the thoracic aorta of rats and were cultured primarily. Real-time PCR was used to measure the mRNA expression of indicated genes. Western blotting was performed to detect the protein expression of autophagy-related markers. We found that treatment with high Pi levels (1 and 3 mM) activated LC3II expression and promoted autophagic flux in VSMCs. Conversely, treatment with an autophagy inhibitor decreased LC3II expression. Pi stimulation dysregulated the expression of several miRNAs such as miR-18a, miR-21, miR-23a, miR-30b, and miR-31a. However, miR-30b overexpression decreased Pi-induced expression of autophagy-related marker genes such as BECN1, ATG5, and LC3b, whereas miR-30b downregulation increased Pi-induced expression of these genes. In addition, we found that miR-30b directly targeted BECN1. These data suggest that miR-30b plays an important role in the regulation of high Pi level-induced autophagy in VSMCs by targeting BECN1.


Case report of hypnic headache: a rare headache disorder with nocturnal symptoms.
Headache is one of the commonest complaints reported to physicians worldwide. Yet, arriving at the proper diagnosis can be a challenge in many patients. Although most headaches belong to common categories of migraine and tension-type headache, which are diagnosed and managed relatively easily, several uncommon headache disorders can lead to delays in diagnosis. Certain medications are more efficacious than others in managing these headache disorders, hence establishing the correct diagnosis is of paramount importance. An 86-year-old female presented with chronic daily headache of 1 year duration. Her headaches were exclusively nocturnal and woke her up daily around midnight. Clinical examination was unremarkable. All basic investigations were normal. Subsequent gadolinium enhanced Magnetic Resonance Imaging (MRI) brain did not show any significant pathology. There was no satisfactory response to paracetamol, diclofenac sodium, mefenamic acid, tramadol, flunarizine and sodium valproate. Indomethacin was started with the provisional diagnosis of hypnic headache. There was absolute response by day 3 of indomethacin. She remains headache free on low dose indomethacin maintenance at 1 year after the diagnosis. Better understanding of uncommon headache syndromes can help in early diagnosis and appropriate treatment. Hypnic headache should be considered in the differential diagnosis of chronic daily headaches, especially when nocturnal and occurs during sleep.


Neglected partial denture in the lower oesophagus presenting after 7 days.
Accidental ingestion of a denture is an acute emergency, and the denture is usually removed on the same day it is discovered. We present a patient who had a seizure while asleep, during which his denture broke; he accidentally swallowed a major part of it, which had a clasp attached. He was unaware that he had ingested the denture, since he was asymptomatic, but he started developing symptoms after 5 days and presented to us on the eighth day of ingestion. With much difficulty, the impacted section of the denture in the distal oesophagus was removed with the combined effort of flexible endoscopy and a rigid oesophagoscopy. Post-procedure, the patient developed mediastinitis, which was managed conservatively; he is doing well on follow-up.


Characterization of a cathepsin D protease from CHO cell-free medium and mitigation of its impact on the stability of a recombinant therapeutic protein.
During purification process development of a recombinant therapeutic protein, an endoproteolytic activity endogenous to the Chinese hamster ovary (CHO) cells and leading to degradation at particular hydrophobic amino acid residues (e.g., Phe and Trp) was observed when processing at acidic pH. The presence of residual levels of protease activity in purified protein batches affected the inherent activity of the product when stored as a solution. To develop a robust purification strategy to minimize this undesirable impact, identification and characterization of this protease was essential to ultimately ensure that a solution formulation was stable for many years. A protease was isolated from CHO cell-free medium (CFM) using a combination of immobilized pepstatin-A agarose chromatography and size exclusion chromatography (SEC). The isolated protease has significant proteolytic activity at pH ∼ 3 to neutral pH and was identified as cathepsin D by mass spectrometry. Analytical SEC, chip-based capillary gel electrophoresis, imaged capillary isoelectric focusing (cIEF), and circular dichroism (CD) spectropolarimetry analyses were performed for additional characterization of the protease. The identification and characterization of this protease enabled the development of a robust purification process by implementation of a controlled temperature inactivation unit operation (heat inactivation) that enabled essentially complete inactivation of the protease, resulting in the production of a stable drug product that had not been possible using column chromatography alone. © 2017 American Institute of Chemical Engineers Biotechnol. Prog., 34:120-129, 2018.


Microbial community in resistant and susceptible Churra sheep infected by Teladorsagia circumcincta.
Gastrointestinal nematodes (GIN) are a major threat to health and welfare in small ruminants worldwide. Teladorsagia circumcincta is a nematode that inhabits the abomasum of sheep, especially in temperate regions, causing important economic losses. Given that T. circumcincta and microbiome share the same niche, interactions between them and the host are expected. Although it is known that within a sheep breed there are animals that are more resistant than others to infection by GIN, it is not known if the microbiome influences the phenotype of these animals. Under this condition, 12 sheep were classified according to their cumulative faecal egg count (cFEC) at the end of a first experimental infection, 6 as resistant group (RG) and 6 as susceptible group (SG) to T. circumcincta infection. Then, all sheep were experimentally infected with 70,000 L3 of T. circumcincta and at day 7 days post-infection were euthanized. At necropsy, gastric mucosa and gastric content from abomasum were collected to extract bacterial DNA and sequence V3-V4 region from 16S rRNA gene using Ilumina technology. After bioanalysis performed, results showed that α-diversity and β-diversity remained similar in both groups. However, resistant phenotype sheep showed a higher number of bacteria butyrate-fermenting species as Clostridium sensu stricto 1 (abundance in RG: 1.29% and in SG: 0.069%; p = 0.05), and Turicibacter (abundance in RG: 0.31% and in SG: 0.027%; p = 0.07) in gastric content but also Serratia spp in gastric mucosa (abundance in RG: 0.12% and in SG: 0.041%; p = 0.07). A trend towards a significant negative correlation between cFEC and Clostridium sensu stricto 1 abundance in gastric content was detected (r = - 0.537; p = 0.08). These data suggest that microbiome composition could be another factor associated with the development of the resistant phenotype modifying the interaction with the host and the in last instance affecting the individual risk of infection.


Ballistic study on the penetration potential and injury potential of different bullet types in the use of a newly developed bullet shooting stunner for adequate stunning of heavy cattle.
Recently, a special bullet shooting stunner for heavy cattle has been developed that fires a bullet instead of a bolt. In the search for a suitable ammunition, the following criteria must be met: First, the energy of the bullet must be sufficient to penetrate the thick frontal bones of heavy cattle. Second, the injury potential at the corresponding penetration depth should preferably be large in order to damage brain tissue relevant to stunning. Third, the bullet must not perforate the occipital bone (over-penetration). Four different bullet types [Hornady FTX, Hydra-Shok, Black Mamba, and a common full metal jacket (FMJ) bullet] were evaluated in a series of experiments on soap blocks and removed bone plates followed by computed tomography examinations. Penetration potential was evaluated in terms of kinetic energy relative to the caliber of the bullet, i.e., mean energy density (ED). Injury potential was evaluated by the mean extent of the cavity volume (e CV ) at the relevant penetration depth of 5.5 to 7.5 cm in the soap block. All four bullet types passed through the frontal bone plate. The ED was 17.50 J/mm2 (Hornady FTX), 17.46 J/mm2 (Hydra-Shok), 13.47 J/mm2 (Black Mamba), and 13.47 J/mm2 (FMJ). The Hornady FTX and the Hydra-Shok each fragmented heavily. The FMJ was excluded after three experiments due to over-penetrations. The e CV was e CV = 3.77 cm2 (Hornady FTX), 2.71 cm2 (Hydra-Shok), and 1.31 cm2 (Black Mamba), with a significant difference (p = 0.006) between the Hornady FTX and the Black Mamba. For use in heavy cattle, the Hornady FTX and the Hydra-Shok are recommended due to the larger e CV than the Black Mamba.


Specific interaction of CD4-bearing liposomes with HIV-infected cells.
The CD4 molecule was reconstituted into the bilayers of large liposomes. Fluorescence microscopy and electron microscopy showed that these liposomes interact with HIV-infected H9-HT cells, delivering their contents to the cell interior. Liposomes bearing CD4 did not interact in this way with noninfected H9-HT cells nor did liposomes without CD4 interact significantly with HIV-infected cells. From electron micrographs, it appeared that HIV binds to liposomes bearing CD4; no attachment of virions to liposomes without CD4 was observed.


Aberrant Expression of Anaplastic Lymphoma Kinase in Ovarian Carcinoma Independent of Gene Rearrangement.
Ovarian carcinoma is the leading cause of death from gynecologic malignancies. The oncogenic role of anaplastic lymphoma kinase (ALK) is well characterized in many hematopoietic and solid tumors. ALK expression in ovarian carcinoma has been reported but the exact status of ALK protein and its association with clinicopathologic features requires further investigation. ALK expression was determined by immunohistochemistry in 110 primary ovarian carcinomas, including 85 cases of serous carcinoma and 25 cases of mucinous carcinoma. Fluorescence in situ hybridization (FISH) and real-time reverse transcription polymerase chain reaction (RT-PCR) were used for evaluating ALK translocation in ALK-positive ovarian carcinomas. Among 110 ovarian carcinomas, 23 (20.9%) cases were ALK positive by immunohistochemistry. All ALK-positive cases were ovarian high-grade serous carcinoma. ALK expression was detected in 23/85 (27.1%) ovarian serous carcinoma and 0/25 (0%) in ovarian mucinous carcinoma. None of the 23 ALK IHC-positive cases harbored ALK gene translocations by FISH or RT-PCR. ALK protein expression was associated with patient age, tumor stage, and histologic type. Specifically, the probability of ALK protein expression was significantly higher in high-grade serous carcinomas in older patients (above 50 y) with advanced disease (FIGO stage III and IV) compared with the low-grade serous and mucinous carcinomas in younger patients with relatively early disease. In conclusion, aberrant ALK expression is observed in ovarian serous carcinoma but not in mucinous carcinoma, is independent of gene translocation, and might be associated with progression and prognosis.


A family with various symptomatology suggestive of Anderson-Fabry disease and a genetic polymorphism of alpha galactosidase A gene.
Anderson/Fabry disease expresses a wide range of clinical variability in patients that it is possible to explain referring to a genetic variability with numerous mutations described in the literature (more than 600). We report some clinical cases of some members of a Sicilian family to express phenotypical variability of this disease in subjects with the same genetic mutation The first case was a 59-year-old female. Brain MRI revealed right frontal periventricular white matter of likely vascular-degenerative origin. The proband's alpha galactosidase A activity was 3.7nmol/mL/h. Molecular genetics revealed a polymorphism: -10 C>T; IVS 2-76_80del5; IVS4-16 A>G; IVS6-22 C>T. The second case was a 30year-old male affected by acroparesthesias and hypoidrosis since he was an adolescent. Renal impairment was first detected at age 29; it began with high plasma levels of creatinine and microalbuminuria date. The third case was a 41year-old daughter that presented with acroparesthesias, hypoidrosis since she was very young. The patient's alpha galactosidase A activity was 4.1nmol/mL/h, in whole blood, which is compatible with heterozygote subject for Fabry's disease or healthy control. The fourth case was a male grandson of the proband, 9year-old child. He had a classic gastrointestinal involvement. He complained of recurrent abdominal pain, post prandial bloating and pain. This child's enzyme activity was 1.65nmol/mL/h. In cases 2, 3, and 4, molecular genetics revealed a polymorphism: -10 C>T; IVS 2-76_80del5; IVS4-16 A>G; IVS6-22 C<T DISCUSSION: A recent study reported that IVS4+68 A>G, IVS6-22C>T polymorphisms occurred in 8.9% and 3.7% of the subjects respectively, and the significance of this haplotype in FD pathology remains unknown but possibly suggestive of Anderson/Fabry disease.


Age at diagnosis of cancer as predictor of mutation occurrence in families suspected of HNPCC.
Analysis of significance of age at cancer diagnosis as a factor allowing identification of a subgroup of patients with a high frequency of hMSH2 and hMLH1 mutations among families that fulfil suspected HNPCC criteria was performed. DNA from thirty-one unrelated patients affected by colorectal cancer from families matching the above criteria were studied by direct sequencing for occurrence of hMSH2 and hMLH1 gene mutations. Seven unequivocal constitutional mutations were detected: five in the hMLH1 gene and two in the hMSH2 gene. Additionally, one hMLH1 alteration of unknown significance was found. All seven mutations were found in a subgroup of 19 patients with cancer diagnosed before the age of 50 years. In a subgroup of 12 patients with cancer diagnosed at an older age only one case with hMLH1 alteration of unknown significance was detected. Our results indicate that early age at cancer diagnosis seems to be a crucial pedigree factor in discrimination of patients with hMSH2 or hMLH1 mutations among families suspected of HNPCC and matching criteria I of ICG-HNPCC.


Evaluation of Disease Severity and Global Transcriptome Response Induced by Citrus bark cracking viroid, Hop latent viroid, and Their Co-Infection in Hop (Humulus lupulus L.).
Viroids are small non-capsidated, single-stranded, covalently-closed circular noncoding RNA replicons of 239-401 nucleotides that exploit host factors for their replication, and some cause disease in several economically important crop plants, while others appear to be benign. The proposed mechanisms of viroid pathogenesis include direct interaction of the genomic viroid RNA with host factors and post-transcriptional or transcriptional gene silencing via viroid-derived small RNAs (vd-sRNAs) generated by the host defensive machinery. Humulus lupulus (hop) plants are hosts to several viroids among which Hop latent viroid (HLVd) and Citrus bark cracking viroid (CBCVd) are attractive model systems for the study of viroid-host interactions due to the symptomless infection of the former and severe symptoms induced by the latter in this indicator host. To better understand their interactions with hop plant, a comparative transcriptomic analysis based on RNA sequencing (RNA-seq) was performed to reveal the transcriptional alterations induced as a result of single HLVd and CBCVd infection in hop. Additionally, the effect of HLVd on the aggressiveness of CBCVd that underlies severe stunting in hop in a mixed infection was studied by transcriptomic analysis. Our analysis revealed that CBCVd infection resulted in dynamic changes in the activity of genes as compared to single HLVd infection and their mixed infection. The differentially expressed genes that are involved in defense, phytohormone signaling, photosynthesis and chloroplasts, RNA regulation, processing and binding; protein metabolism and modification; and other mechanisms were more modulated in the CBCVd infection of hop. Nevertheless, Gene Ontology (GO) classification and pathway enrichment analysis showed that the expression of genes involved in the proteolysis mechanism is more active in a mixed infection as compared to a single one, suggesting co-infecting viroids may result in interference with host factors more prominently. Collectively, our results provide a deep transcriptome of hop and insight into complex single HLVd, CBCVd, and their coinfection in hop-plant interactions.


The human rosette-forming cell as a marker of a population of thymus-derived cells.
Sheep red blood cells can surround, in vitro, some human peripheral blood lymphocytes in a formation called a rosette. The number of rosetteforming cells (RFC) in 50 normal persons had a wide range (4-40%). The organs of 13 human fetuses (11-19 wk conceptional age) were examined for the presence of RFC. The thymus possessed the highest percentage of RFC, the maximum being 65% of total thymocytes in two 15-16 wk fetal specimens. Blood RFC were always present and their number slightly increased in the oldest fetuses. The bone-marrow showed 0-8% in the six fetuses studied. RFC were found in the spleen around the 13th wk and in the liver around the 17th wk of gestation. These observations lead to the hypothesis that human blood RFC may be chiefly thymic derived. Studies of patients with immunological disorders support this hypothesis: one patient with Nezelof syndrome had no blood RFC and four patients with Wiskott-Aldrich syndrome had a low number of blood RFC (1 and 1.5%). Patients with acquired hypogammaglobulinemia showed a normal percentage of RFC. With the fetal thymocytes, the percentage of inhibition with anti-mu serum increased with the fetal age to become complete in the oldest fetuses studied. Incubation of the oldest fetal thymocytes or the blood lymphocytes with anti-gamma serum of anti-mu serum completely inhibited the rosette formation. These results suggest that mu-chain determinants are present on human fetal thymocytes and blood RFC. The significance of the presence of gamma-chain determinants on these cells is unclear.


Patients with high-grade gliomas harboring deletions of chromosomes 9p and 10q benefit from temozolomide treatment.
Surgical cure of glioblastomas is virtually impossible and their clinical course is mainly determined by the biologic behavior of the tumor cells and their response to radiation and chemotherapy. We investigated whether response to temozolomide (TMZ) chemotherapy differs in subsets of malignant glioblastomas defined by genetic lesions. Eighty patients with newly diagnosed glioblastoma were analyzed with comparative genomic hybridization and loss of heterozygosity. All patients underwent radical resection. Fifty patients received TMZ after radiotherapy (TMZ group) and 30 patients received radiotherapy alone (RT group). The most common aberrations detected were gains of parts of chromosome 7 and losses of 10q, 9p, or 13q. The spectrum of genetic aberrations did not differ between the TMZ and RT groups. Patients treated with TMZ showed significantly better survival than patients treated with radiotherapy alone (19.5 vs 9.3 months). Genomic deletions on chromosomes 9 and 10 are typical for glioblastoma and associated with poor prognosis. However, patients with these aberrations benefited significantly from TMZ in univariate analysis. In multivariate analysis, this effect was pronounced for 9p deletion and for elderly patients with 10q deletions, respectively. This study demonstrates that molecular genetic and cytogenetic analyses potentially predict responses to chemotherapy in patients with newly diagnosed glioblastomas.


Effect of serotonin on ovulation in the fowl.
Serotonin (5-hydroxytryptamine) injected 8 hours before the expected time of ovulation effectively blocked ovulation (90%). When serotonin was administered for 10 days, the ovarian weight number follicles, adrenal weights, and ovulation rate were significantly changed (p less than or equal to .05). Serotonin or reserpine treatment induced premature oviposition if the egg was in the genital tract. It was suggested from this study that there is stimulation for ovulation at 8 hour before ovulation which can be inhibited by serotonin.


Trigeminal Neuralgia in Pregnancy: A Management Challenge.
Pregnancy is known to aggravate pre-existing chronic painful conditions. Trigeminal neuralgia (TN), albeit a disease of the elderly, may afflict pregnant females, which can further complicate its management. Teratogenic effects of the commonly used drugs on the developing fetus limit pharmacological treatment. Moreover, safety of commonly performed interventional therapies is marred by their inherent fetomaternal effects and more importantly the risk for radiation effects on the fetus due to the use of fluoroscopy. This rare coexistence of TN in pregnancy has not been reported before. Here we present a case of TN in a young woman, whose pain was aggravated when she became pregnant, and she was treated successfully by conventional radiofrequency ablation of the Gasserian ganglion.


Wheelchair Neuroprosthesis for Improving Dynamic Trunk Stability.
Trunk instability is a major problem for individuals with thoracic and cervical spinal cord injury. Functional electrical stimulation (FES) neuroprosthesis, a technology that uses small electrical currents to artificially contract muscles, has previously been utilized to improve trunk stability during quasi-static and dynamic sitting. The aim of this paper was to develop the first powered wheelchair-based neuroprosthesis and to test its feasibility for improving trunk stability. Eleven male, able-bodied individuals participated in the feasibility study. While participants were seated, the wheelchair was moved in the forward or backward directions with slow and fast accelerations. Two different FES protocols were tested: 1) co-contraction and 2) directionally-dependent contraction of trunk extensors and flexors. Sham stimulations with intensities below the motor threshold were applied as the control conditions. Inertial motion sensors were used to quantify the maximum angular displacement and velocity of the trunk. Results showed that both directional contractions and co-contraction reduced trunk displacement and velocity, compared with the control conditions. However, directionally-dependent muscle contractions were more effective in improving trunk stability, compared with co-contractions. Overall, feasibility of the wheelchair-based neuroprosthesis was demonstrated. Future research will incorporate feedback from wheelchair movements and test the neuroprosthesis with individuals who sustained spinal cord injury.


Medication Adherence in Adolescents with Psychiatric Disorders.
Objective: This study investigates whether adolescents' adherence to psychotropic medication is associated with demographic and socioeconomic factors, and to what extent parents' assessments of their offspring's attitudes toward treatment correspond with the adolescents' self-report. Methods: This study is part of the multicenter SEMA study (Subjective Experience and Medication Adherence in Adolescents with Psychiatric Disorders). Adolescents' subjective attitudes toward medication and their adherence were assessed using the patient and parent versions of the QATT (Questionnaire on Attitudes Toward Treatment) and the MARS (Medication Adherence Rating Scale). Furthermore, we collected socioeconomic and demographic data. Results: Of the n = 75 adolescents included in the study, n = 45 (60 %) were classified as completely adherent. Patients receiving monotherapy were more often completely adherent than those receiving a combination of different medications. There was no statistically significant association between adherence and demographic or socioeconomic factors. Consensus between adolescents and their parents regarding adolescents' attitudes toward treatment ranged from slight (κ = 0.157) to fair (κ = 0.205). Conclusion: Incomplete medication adherence in adolescents with psychiatric disorders is a common phenomenon and still poorly understood. Demographic and socioeconomic factors do not seem to be relevant in this respect. However, adolescents' subjective attitudes towards medication, which parents are presumably unable to adequately assess, warrant more careful consideration in future research.


Stronger declines in youth alcohol consumption thanks to stronger integrated alcohol policies? A qualitative comparison of ten Dutch municipalities.
Little detailed evidence is available on how integrated policies could impact population health and under what conditions such policies could be realized. The aim of this study was to assess how youth alcohol consumption trends in the province of Noord-Brabant, The Netherlands, were related to the development and implementation of integrated policies. In a retrospective multiple case study, alcohol policies of six municipalities with stronger declines in youth alcohol consumption between 2007 and 2011 (cases) were compared to four municipalities with weaker declines (controls). Information on the policy process in the same period was obtained through semi-structured in-depth interviews with policy advisors. Information on implemented interventions was extracted from policy documents and checked by the interviewees. Interviews were analyzed for thematic content. Only municipalities with stronger declines in alcohol consumption involved sectors other than public health and had started to implement interventions that use regulatory or enforcement strategies. Their involvement was facilitated by framing youth alcohol consumption as a safety rather than a health problem, whereby local media played a substantial role. Implementation of integrated policies was further facilitated by dedicated leadership and sufficient resources. Reductions in youth alcohol consumption in Noord-Brabant were stronger when municipalities started to develop integrated policies. Results suggest that integrated policies framing a health problem as a broader societal problem could positively influence population health.


Aberrant splicing events that are induced by proviral integration: implications for myb oncogene activation.
Activation of the mouse c-myb oncogene in Abelson virus-induced plasmacytoid lymphosarcomas was studied using cDNA cloning and nucleotide sequence analysis. The results presented here show that viral integration in the myb locus generates splicing errors at the 5' and 3' regions. Viral integration results in transcriptional initiation within the viral long terminal repeat and generation of a chimeric mRNA that lacks the first three coding exons. The alterations at the 3' end are caused by an aberrant splicing event in which additional splice-donor and -acceptor sequences within intronic sequences are used to splice an additional 363 nucleotides into the myb transcripts. The resulting insertion of 121 amino acids is in a region of the protein where other activated forms of the myb gene product have deletions. These results suggest that alterations in the 3' end of the myb gene play a crucial role in the activation of this gene.


Synthesis and antitumor activity of 10-propargyl-10-deazaaminopterin.
Successive alkylation of dimethyl homoterephthalate with propargyl bromide and 2,4-diamino-6-(bromomethyl)pteridine followed by ester saponification at room temperature afforded 2,4-diamino-4-deoxy-10-carboxy-10-propargyl-10-deazapteroic acid. The 10-COOH was readily decarboxylated by heating in DMSO at a temperature of only 120 degrees C to yield the diamino-10-propargyl-10-deazapteroic acid intermediate. Coupling with diethyl L-glutamate and ester hydrolysis gave the title compound. The 10-propargyl analogue was about 5 times more potent than MTX as an inhibitor of growth in L1210 cells, but was only one-third as potent as an inhibitor of DHFR from L1210. The analogue was transported inward very effectively in L1210 cells showing a 10-fold advantage over MTX. At a dose of 36 mg/kg the 10-propargyl compound caused shrinkage of the E0771 solid murine mammary tumor to only 1% of untreated controls.


Diastereomeric Xe chemical shifts in tethered cryptophane cages.
Cryptophane cages serve as host molecules to a Xe atom. Functionalization of cryptophane-A has permitted the development of Xe as a biosensor. Synthetic routes used to prepare cryptophanes result in racemic mixtures of the chiral cages. In the preparation of a tethered cryptophane-A cage for biosensor applications, some achiral and chiral substituents such as left-handed amino acids have been used. When the substituent is achiral, the NMR signal of the Xe atom in the functionalized cage in solution is a single isotropic peak, since the Xe shielding tensor components in the R and L cages differ by no more than the signs of the off-diagonal elements. Chiral substituents can split the cage-encapsulated Xe NMR signal into one or more sets of doublets, depending on the number of asymmetric centers in the substituent. We carry out quantum mechanical calculations of Xe nuclear magnetic shielding for the Xe atom at the same strategic position within an L cryptophane-A cage, under the influence of chiral potentials that represent r or l substituents outside the cage. Calculations of the Xe shielding response in the Lr and Ll diastereomeric pairs permit the prediction of the relative order of the Xe chemical shifts in solutions containing the Rl and Ll diastereomers. Where the substituent itself possesses two chiral centers, comparison of the calculated isotropic shielding responses in the Llr, Lrl, Rll, and Lrr systems, respectively, permits the prediction of the Xe spectrum of diastereomeric systems in solutions containing Llr, Rlr, Lll, and Rll systems. Assignment of the peaks observed in the experimental Xe NMR spectra is therefore possible, without having to undertake the difficult synthetic route that produces a single optically pure enantiomer.


The quest for new innovative care delivery models.
In response to growing nurse shortages and heightened attention to patient quality, many healthcare organizational leaders are developing and testing new care delivery models. These models strive to improve patient quality and satisfaction by engaging nurses and other healthcare professionals in different roles across the continuum of care. In this article, the authors profile 5 new care delivery models from their current work sponsored by the Robert Wood Johnson Foundation. In addition, the authors identify common elements underlying the success of the models.


Comparative study on physicochemical properties of starch films prepared from five sweet potato (Ipomoea batatas) cultivars.
Five different sweet potato (Ipomoea batatas) cultivars (Daeyumi, Gogeonmi, Sincheonmi [SCM], Singeonmi, and Sinyulmi [SYM]) were used to extract sweet potato starch (SPS) for developing starch-based films. After the chemical composition and amylose contents of all SPSs were evaluated, the morphological, moisture, mechanical, and barrier properties of the SPS-based films were investigated. As one of the film characteristics, the X-ray diffractograms revealed that the SCM-based film with the highest amylose content (26.34%) had the highest relative crystallinity (24.31%). The SCM-based film also showed higher tensile strength (3.05-fold) and elastic modulus (2.38-fold) than the SYM-based film with the lowest amylose content (21.84%). The water vapor and oxygen permeabilities of the SPS-based films were negatively correlated with the amylose content. Thus, the SCM-based film was less permeable for water vapor (3.16-fold) and oxygen (1.81-fold) than the SYM-based film. These results demonstrated that the sweet potato cultivar, especially the amylose content, plays a significant role in determining the physicochemical properties of the SPS-based films.


Loss of DNA mismatch repair function and cancer predisposition in the mouse: animal models for human hereditary nonpolyposis colorectal cancer.
Germline mutations in DNA mismatch repair genes underlie one of the most common hereditary cancer predisposition syndromes known in humans, hereditary nonpolyposis colorectal cancer (HNPCC). Defects of the DNA mismatch repair system are also prevalent in sporadic colorectal cancers. The generation of mice with targeted inactivating mutations in the mismatch repair genes has facilitated the in vivo study of how these genes function and how their individual loss contributes to tumorigenesis. Although there are notable limitations when using murine models to study the molecular basis of human cancer, there is remarkable similarity between the two species with respect to the contribution of individual members of the mismatch repair system to cancer susceptibility, and mouse mutants have greatly enhanced our understanding of the normal role of these genes in mutation avoidance and suppression of tumorigenesis.


Data on the exon-intron organization of genes coding for B-cell receptor-like proteins.
B-cell receptor-associated protein (BAP) family plays important roles in the ER homeostasis and stress responses of eukaryotic cells [1]. We reported the analysis of plant BAP-like (PBL) genes and the encoded proteins of higher land plants [2]. The origin and functional divergence of these genes among all eukaryotes, however, are poorly studied, which impedes our understanding of the functional relationships and diversity among BAP-like proteins. One possible reason for the potential functional diversity may be the differences in the exon-intron structure of PBL genes. In this study, we first performed analysis of the exon-intron organization of these genes in the genome sequences of the Viridiplantae species in addition to previously published data on Angiosperms [2]. To further address the distribution of BAP-like genes in other eukaryotes, we extended our dataset to include the representative genes encoded by non-plant bikonts and unikonts [3].


CRISPR/Cas9-Mediated SlNPR1 mutagenesis reduces tomato plant drought tolerance.
NPR1, nonexpressor of pathogenesis-related gene 1, is a master regulator involved in plant defense response to pathogens, and its regulatory mechanism in the defense pathway has been relatively clear. However, information about the function of NPR1 in plant response to abiotic stress is still limited. Tomato is the fourth most economically crop worldwide and also one of the best-characterized model plants employed in genetic studies. Because of the lack of a stable tomato NPR1 (SlNPR1) mutant, little is known about the function of SlNPR1 in tomato response to biotic and abiotic stresses. Here we isolated SlNPR1 from tomato 'Ailsa Craig' and generated slnpr1 mutants using the CRISPR/Cas9 system. Analysis of the cis-acting elements indicated that SlNPR1 might be involved in tomato plant response to drought stress. Expression pattern analysis showed that SlNPR1 was expressed in all plant tissues, and it was strongly induced by drought stress. Thus, we investigated the function of SlNPR1 in tomato-plant drought tolerance. Results showed that slnpr1 mutants exhibited reduced drought tolerance with increased stomatal aperture, higher electrolytic leakage, malondialdehyde (MDA) and hydrogen peroxide (H2O2) levels, and lower activity levels of antioxidant enzymes, compared to wild type (WT) plants. The reduced drought tolerance of slnpr1 mutants was further reflected by the down-regulated expression of drought related key genes, including SlGST, SlDHN, and SlDREB. Collectively, the data suggest that SlNPR1 is involved in regulating tomato plant drought response. These results aid in further understanding the molecular basis underlying SlNPR1 mediation of tomato drought sensitivity.


Evaluation of the efficacy of green tea catechins as an adjunct to scaling and root planing in the management of chronic periodontitis using PCR analysis: A clinical and microbiological study.
Increased knowledge of anaerobic bacteria in the development of periodontal diseases has led to new treatment strategies aiming primarily at suppression or elimination of specific periodontal pathogens. Over the last few decades, green tea has been subjected to many scientific and medical studies. The purpose of the present study was to assess the effect of green tea catechins on the red complex organisms using Polymerase Chain Reaction for microbiological analysis. A split mouth study was conducted, in which a total of 20 subjects were included. Green tea catechin as local drug delivery was placed at study sites. Clinical parameters namely probing pocket depth (PPD), gingival index (GI), plaque index (PI) were recorded. Sub-gingival plaque samples were collected, and red complex micro-organisms were studied using PCR. Clinical and microbiological parameters were recorded at baseline, 1(st), and 5(th) week after treatment. The results showed statistically significant difference in PPD, GI, and PI and significant reduction of red complex organisms from baseline to 1(st) week and baseline to 5(th) week in both study and control groups (P < 0.001). Intergroup comparison between study and control group was statistically insignificant for PPD, PI, and GI. A significantly greater reduction in Tannerella forsythus (Tf) at 1(st) week and 5(th) week and Porphyromonas gingivalis (Pg) at 1(st) week was observed in study group when compared to control group. Green tea catechin can be used as an effective local drug delivery along with scaling and root planing in treatment of chronic periodontitis.


The Levels of Sera Malondialdehyde, Erythrocyte Membrane Na+-K+/Mg++ and Ca++/Mg++ Adenosine 5' Triphosphatase in Patients with Sickle Cell Anemia.
The various membrane abnormalities of sickle erythrocytes may result from excessive accumulation of oxidant damage. We measured the sera levels of malondialdehyde, products of lipid peroxidation, Na+-K+/Mg++ Adenosine 5' triphosphatase (ATPase) and Ca++/Mg++ Adenosine 5' triphosphatase, erythrocyte membrane enzymes, in patients with sickle cell anemia and compared their levels with that of normal controls. MDA, Na+-K+/Mg++ and Ca++/Mg++ ATPase levels of control groups were 0.90 ± 0.04 nmol/mL, 168 ± 12.9 and 140.6 ± 8.2 nmol Pi/mg prot/hour, respectively. MDA, Na+-K+/Mg++ and Ca++/Mg++ATPase levels of patients were 2.02 ± 0.01 nmol/mL, 127.0 ± 8.4 and 110.0 ± 9.6 nmol Pi/mg prot/hour, respectively. Our experimental results showed that there was a significant increase in MDA levels of patients with sickle cell anemia as compared with that of the controls. However, erythrocyte membrane Na+-K+/Mg++ and Ca++/Mg++ ATPase levels of patients with sickle cell anemia were significantly lower than the, Na+-K+/Mg++ and Ca++/Mg++ ATPase levels of normal controls.


Preferences for Attention-Deficit/Hyperactivity Disorder (ADHD) Non-Stimulant Treatment Characteristics Among Children and Adolescents With ADHD and Their Caregivers.
Background: Understanding patient and caregiver preferences for treatment is important for optimizing treatment decisions. Non-stimulant therapies are an alternative treatment option to stimulant therapy for attention-deficit/hyperactivity disorder (ADHD). Guanfacine extended release (GXR) and atomoxetine (ATX) are two non-stimulant medications approved in the United States for the treatment of ADHD. Objective: To identify non-stimulant ADHD medication attributes important to caregivers/patients. Methods: US caregivers of ADHD patients (6-17 years) and child/adolescent patients (10-17 years) completed an adaptive conjoint analysis survey. Respondents selected between hypothetical treatments with different attributes. Ordinary least-squares and hierarchical Bayes regression using Sawtooth Software were used to calculate utilities, importance ratings, and preferences. Results: 483 caregivers (mean age: 41.9 years, standard deviation [SD]: 8.7; 75% female) and 211 children/adolescents (mean age: 14.5 years, SD: 2.2; 70% male) completed the survey. Based on importance ratings, the most influential attributes for both caregivers and children/adolescents were chance of somnolence, efficacy, and for caregivers, effect on oppositionality and black box warning. Most caregivers (95.3%) and children/adolescents (93.8%) preferred GXR over ATX. In several sensitivity analyses in which attribute levels varied, GXR remained the preferred medication with the exception of one scenario. Conclusions: Children/adolescents and caregivers demonstrated in this study that they can clearly express their preferences for treatment attributes and treatment choices; in this case they preferred GXR to ATX. Patients and caregiver preferences could be useful inputs to the treatment selection decision-making process.


Taurine prevents streptozotocin impairment of hormone-stimulated glucose uptake in rat adipocytes.
Streptozotocin-treated rats were used as models of type 1 diabetes to study the effects of dietary taurine on insulin- and adrenergic-stimulated 2-deoxyglucose uptake by isolated adipocytes. In addition to the well-established impairment of basal and insulin-stimulated 2-deoxyglucose uptakes in adipocytes prepared from streptozotocin-diabetic rats, the alpha-(phenylephrine) and beta-(isoproterenol) adrenergic stimulations of glucose uptake were also abolished. The insulin stimulation of glucose uptake in adipocytes was selectively abolished by the phosphatidylinositol 3-kinase inhibitor wortmannin, whereas that by the adrenergic agonists, phenylephrine and isoproterenol, was inhibited by prazosin and propranolol, respectively. Dietary taurine, 4 weeks before and 4 weeks after streptozotocin administration, prevented the loss of both insulin and adrenergic agonist stimulations of 2-deoxyglucose uptake, without affecting hyperglycaemia. Because insulin and adrenergic activations of glucose transport by adipocytes are coupled to different signalling pathways, it is unlikely that these effects of taurine are related to these disparate postreceptor mechanisms.


[Proprotein Convertase Subtilisin/Kexin-Type 9 (PCSK9) New Opportunities of Lipid-Lowering Therapy].
In the literature review covered issues opening protein-proprotein convertase, subtilisin/kexin-type9 (PCSK9), its modern terminology, the results of its biochemical, molecular and genetic studies, metabolic regulation, functions and clinical findings in the blood content ofPCSK9 in lipid disorders and clinical pharmacological studies of monoclonal antibodies to this protein for the correction of lipid metabolism of major interest for cardiology and lipidology.


Real-time monitoring of epithelial cell-cell and cell-substrate interactions by infrared surface plasmon spectroscopy.
The development of novel technologies capable of monitoring the dynamics of cell-cell and cell-substrate interactions in real time and a label-free manner is vital for gaining deeper insights into these most fundamental cellular processes. However, the label-free technologies available today provide only limited information on these processes. Here, we report a new (to our knowledge) infrared surface plasmon resonance (SPR)-based methodology that can resolve distinct phases of cell-cell and cell-substrate adhesion of polarized Madin Darby canine kidney epithelial cells. Due to the extended penetration depth of the infrared SP wave, the dynamics of cell adhesion can be detected with high accuracy and high temporal resolution. Analysis of the temporal variation of the SPR reflectivity spectrum revealed the existence of multiple phases in epithelial cell adhesion: initial contact of the cells with the substrate (cell deposition), cell spreading, formation of intercellular contacts, and subsequent generation of cell clusters. The final formation of a continuous cell monolayer could also be sensed. The SPR measurements were validated by optical microscopy imaging. However, in contrast to the SPR method, the optical analyses were laborious and less quantitative, and hence provided only limited information on the dynamics and phases of cell adhesion.