Comparison of blood-brain barrier disruption by intracarotid iopamidol and methylglucamine iothalamate.
Using a canine model, the effect of intracarotid injections of the ionic contrast medium methylglucamine iothalamate was compared with that of the nonionic contrast medium iopamidol of similar iodine concentration (280 mg 1/ml). The degree and distribution of blood-brain barrier disruption was assessed using Evans blue stain as a visual marker and by contrast enhancement measured by a computed tomographic (CT) scanner. In all studies with methylglucamine iothalamate, Evans blue staining was demonstrated, and CT enhancement demonstrated a significant mean difference (p less than 0.01) between the control and injected hemispheres. The absence of blood-brain barrier disruption with iopamidol is probably related to its lower osmolality (570 mosmol/kg) compared with methylglucamine iothalamate (1,424 mosmol/kg) and the absence of any cation.


The characteristics of cardiac vagal preganglionic motoneurones in the dogfish.
Preganglionic vagal motoneurones supplying the heart of the dogfish have been located in the medulla by antidromic stimulation of the central cut end of the branchial cardiac branch of the vagus. They supplied axons with conduction velocities between 4.75 and 16.3 m s-1, which is similar to mammalian B fibres. Motoneurones were found in two locations: the rostromedial (N = 5) and lateral (N = 12) divisions of the vagal motor column. Their measured depths and rostrocaudal distributions with respect to obex corresponded with the location of branchial cardiac motoneurones determined by horseradish peroxidase (HRP) histochemistry. All the neurones located in the rostromedial division of the vagal motor column were spontaneously rhythmically active. Their activity contributed to the rhythmic, respiratory-related bursts in peripheral recordings of efferent activity from the branchial cardiac vagus. They could be induced to fire in a prolonged burst by mechanical stimulation of the gill arches. The neurones located lateral to the rostromedial division of the vagal motor column could be divided into three categories: (1) spontaneously, continuously active cells which could be induced to fire more frequently by mechanoreceptor stimulation, (2) silent cells which could be induced to fire by mechanoreceptor stimulation, (3) silent cells which did not respond to mechanoreceptor stimulation. It is concluded, from the response of the medial and two categories of lateral cells to mechanoreceptor stimulation (which results in a transient bradycardia), that branchial cardiac motoneurones from both these central locations exert a chronotropic influence on the heart.


Ulcerative Colitis Diagnosed through Evaluation of Underlying Diseases in a Pyoderma Gangrenosum Adolescent without Gastrointestinal Symptoms.
Pyoderma gangrenosum (PG) is a rare, non-infectious, neutrophilic dermatosis characterized by painful ulcers with indistinct borders and peripheral erythema. The diagnosis of PG requires the exclusion of other causes of similar appearing skin manifestations, including vasculitis and infections. The pathogenesis of PG is not clear; however, dysregulation of the immune system has been suggested in previous studies. More than half of the PG patients have underlying diseases; the most common being inflammatory bowel disease (IBD). The progression of PG in IBD patients is seen after the onset of IBD, usually during its exacerbation. On the other hand, PG may follow a course independent of the intestinal disease. We present a case of an 18-year-old young male with PG that presented before being diagnosed with ulcerative colitis as an associated condition. He had a painful ulcerative lesion on his right shin with no previous gastrointestinal symptoms. This case suggests that investigating for underlying disorders is essential in PG patients despite the lack of symptoms other than the skin lesions.


Molecular epidemiology of clinically significant antibiotic resistance genes.
Antimicrobials were first introduced into medical practice a little over 60 years ago and since that time resistant strains of bacteria have arisen in response to the selective pressure of their use. This review uses the paradigm of the evolution and spread of beta-lactamases and in particular beta-lactamases active against antimicrobials used to treat Gram-negative infections. The emergence and evolution particularly of CTX-M extended-spectrum beta-lactamases (ESBLs) is described together with the molecular mechanisms responsible for both primary mutation and horizontal gene transfer. Reference is also made to other significant antibiotic resistance genes, resistance mechanisms in Gram-negative bacteria, such as carbepenamases, and plasmid-mediated fluoroquinolone resistance. The pathogen Staphylococcus aureus is reviewed in detail as an example of a highly successful Gram-positive bacterial pathogen that has acquired and developed resistance to a wide range of antimicrobials. The role of selective pressures in the environment as well as the medical use of antimicrobials together with the interplay of various genetic mechanisms for horizontal gene transfer are considered in the concluding part of this review.


Behavior in preschool children with the 22q11.2 deletion syndrome.
Children with the 22q11.2 deletion syndrome (22q11DS) are at an increased risk of psychiatric problems from pre-adolescence; little is known, however, about behavioral problems at a preschool age and the relationship between speech and behavior in this group. Parents of 90 children (aged 1.42-5.99 years) with 22q11DS filled out the Child Behavior Checklist, documenting behaviors including speech problems. Their profiles were compared with those of a comparison group consisting of 33 children with nonsyndromic orofacial clefts without 22q11DS, since both children with 22q11DS and children with clefts are expected to have speech problems. In the 22q11DS group, data on intelligence was acquired by means of formal tests. Parents of children with 22q11DS reported significantly higher mean scores on withdrawn behavior, affective problems and pervasive developmental problems compared to children with nonsyndromic clefts. Approximately 30% of children with 22q11DS had a score above the 97th percentile on at least one of the behavior subscales, indicating psychopathology. In children with 22q11DS, the reported behavioral problems were not associated with speech problems. Behavioral problems were found in 30% of young children with 22q11DS and were unlikely to be caused by speech problems. Within the 22q11DS group, behavioral problems were not related to the degree of cognitive impairment. This shows that many children with 22q11DS, known to be at an increased risk of psychiatric problems from pre-adolescence, already show behavioral problems before the age of 6 years.


Fecal calprotectin use in inflammatory bowel disease and beyond: A mini-review.
Given the number of inflammatory disorders affecting the gastrointestinal tract directly and indirectly, coupled with the considerable overlap with functional disorders, it is evident that more useful noninvasive diagnostic tests are required to aid with diagnosis. If these tests can also have some utility for individual patient follow-up in terms of disease activity and response to treatment, as well as providing forewarning of disease relapse, it would be extremely useful information for the clinician. One recently described test that may fulfill several of these attributes is based on leakage of a mononuclear cell cytoplasmic protein, calprotectin, along the intestinal tract, which can then be quantified in feces. This has been used to distinguish patients exhibiting symptoms of irritable bowel syndrome from patients with inflammatory bowel disease, with a measure of success greater than with currently used techniques. The present article summarizes the experience with this test used in inflammatory bowel disease, as well as a variety of gastrointestinal disorders.


Hemosuccus pancreaticus following Parathyroidectomy for Primary parathyroid adenoma.
Hemosuccus pancreaticus is defined as intermittent bleeding from the pancreatic duct into the GI tract, usually caused by the rupture of an pseudoaneurysm, which is usually associated with pancreatitis, abdominal surgery, and can result in life-threatening hemorrhage. Hyperparathyroidism is a rare cause of pancreatitis, it should be suspected in all patients with a history of primary hyperpara¬t hyroidism, When these diseases coexist, the course of pancreatitis can be rather severe and often complicated. This report detail a case of Hemosuccus pancreaticus due to hyperparathyroidism associated pancreatitis and its successful emergency conservative management by Embolization Techniques.


[Directed trees and its application to social sciences].
Directed trees are special graphs usually employed as illustrative device of numerical methods applied in evolutionary biology. The present note explores its possible application in social sciences. Its utility as classificatory method in a classical anthropological study of cultural exchanges is exemplified.


Aesthetic management of the breast following explantation: evaluation and mastopexy options.
This retrospective study reviews 720 patients referred for evaluation of their silicone gel-filled breast implants from December of 1992 to January of 1996. Of the 720 patients evaluated, 282 (39.2 percent) subsequently underwent explantation, and 59 of these patients (20.9 percent) had a breast contouring procedure performed at the time of explantation. Our definition of explantation is the operative removal of the implant as well as the implant capsule. The overall complication rate for explantation was 5 out of 282 patients (1.8 percent), whereas the rate of complication among the patients who underwent simultaneous breast contouring was 2 out of 59 patients (3.4 percent). This article presents the management of the breast following explantation, implant removal, and capsulectomy. We review both the preoperative assessment of patients seeking explantation and our technique of explantation. Additionally, we address the importance of preoperative breast ptosis in technique selection and have developed a practical clinical algorithm for guiding simultaneous explantation and breast contouring. We also identify those patients who should undergo delayed breast contouring due to associated risk factors (smoking, need for > 4 cm of nipple movement, and paucity of breast parenchyma).


Assessing the exacerbations risk of influenza-associated chronic occupational asthma.
The purpose of this article was to conduct a risk-based study based on a linkage of experimental human influenza infections and fluctuation analysis of airway function to assess whether influenza viral infection was risk factor for exacerbations of chronic occupational asthma. Here we provided a comprehensive probabilistic analysis aimed at quantifying influenza-associated exacerbations risk for occupational asthmatics, based on a combination of published distributions of viral shedding and symptoms scores and lung respiratory system properties characterized by long-range peak expiratory flow (PEF) dynamics. Using a coupled detrended fluctuation analysis-experimental human influenza approach, we estimated the conditional probability of moderate or severe lung airway obstruction and hence the exacerbations risk of influenza-associated occupational asthma in individuals. The long-range correlation exponent (alpha) was used as a predictor of future exacerbations risk of influenza-associated asthma. For our illustrative distribution of PEF fluctuations and influenza-induced asthma exacerbations risk relations, we found that the probability of exacerbations risk can be limited to below 50% by keeping alpha to below 0.53. This study also found that limiting wheeze scores to 0.56 yields a 75% probability of influenza-associated asthma exacerbations risk and a limit of 0.34 yields a 50% probability that may give a representative estimate of the distribution of chronic respiratory system properties. This study implicates that influenza viral infection is an important risk factor for exacerbations of chronic occupational asthma.


[Palliative treatment of primary lung tumors with transpulmonary chemoembolization (TPCE)].
To evaluate transpulmonary chemoembolization (TPCE) as a symptomatic palliative method for treating inoperable primary lung tumors. From 2002 to 2005, 17 patients (17 males, 3 females; average age: 64.5 years) suffering from primary lung tumors were treated in 3.6 sessions (range: 2 to 8) using TPCE. The patients had the following primary tumors: adenocarcinoma (n = 6), pleural mesothelioma (n = 2), squamous cell carcinoma (n = 1), small cell carcinoma (n = 1), and non-small cell carcinoma (n = 7). After femoral vein puncture, tumor-supplying pulmonary arteries were selectively explored, and 5 - 10 mg mitomycin C and 5 - 10 mL lipiodol and microsphere particles (Spherex) (20 - 70 microm in diameter) were applied with balloon protection. Diagnosis and follow-up were performed in 4-week intervals with unenhanced and contrast-enhanced computed tomography (CT). The mean follow-up was 11.3 months. Treatment was well tolerated by all patients with no major side effects or complications. The laboratory parameters were not significantly influenced. 11.8 % of the patients (n = 2) showed high or moderate lipiodol uptake, and 76.5 % (n = 13) showed low lipiodol uptake. After evaluation of morphologic criteria, a mean volume regression of 12.1 ml (40.4 %) of the embolized areas was achieved in four patients (23.5 %), while a constant value was identified during follow-up for seven patients (41.2 %). In six patients (35.3 %), progression of the treated lung tumors was recorded. The tumor increased by a mean of 38.37 ml (165.38 %). TPCE is a well-tolerated palliative treatment option for patients with primary lung tumors.


Early onset demyelinating Charcot-Marie-Tooth disease caused by a novel in-frame isoleucine deletion in peripheral myelin protein 2.
Peripheral myelin protein 2 (PMP2) is a small protein located on the cytoplasmic side of compact myelin, involved in the lipids transport and in the myelination process. In the last years few families affected with demyelinating Charcot-Marie-Tooth neuropathy (CMT1), caused by PMP2 mutations, have been identified. In this study we describe the first case of a PMP2 in-frame deletion. PMP2 was analyzed by direct sequencing after exclusion of the most frequent CMT-associated genes by using a next generation sequencing (NGS) genes panel. Sanger sequencing was used for family's segregation analysis. Molecular modeling analysis was used to evaluate the mutation impact on the protein structure. A novel PMP2: p.I50del has been identified in a child with early onset CMT1 and in three affected family members. All family members show an early onset demyelinating neuropathy without other distinguish features. Molecular modeling analysis and in silico evaluations do not suggest a strong impact on the overall protein structure, but a most likely altered protein function. This study suggests the importance to add PMP2 in CMT NGS genes panels or, at most, to test it after major CMT1 genes exclusion, due to the lack of diagnostic-addressing additional features.


A general, versatile and divergent synthesis of selectively deuterated amines.
Deuterated organic molecules are of utmost importance in many areas of science and have been recently intensively investigated in medicinal chemistry due to their enhanced metabolic stability. The development of efficient and broadly applicable methods for the selective incorporation of deuterium atoms into organic molecules from readily available starting materials and reagents is therefore of extreme importance. Such methods however often lack generality and selectivity, notably in the nitrogen series. With nitrogen-containing molecules being indeed ubiquitous in medicinal chemistry, there is a strong need for efficient methods enabling the selective synthesis of deuterated amines. In this perspective, we report herein a general, versatile, divergent and metal-free synthesis of amines selectively deuterated at their α and/or β positions. Upon simple treatment of readily available ynamides with a mixture of triflic acid and triethylsilane, either deuterated or not, a range of amines can be smoothly obtained with high levels of deuterium incorporation by a unique sequence involving a domino keteniminium/iminium activation.


Bimanual coordination in alcohol-exposed children: role of the corpus callosum.
The corpus callosum (CC) is one of several brain structures affected in children prenatally exposed to alcohol. This structure plays a major role in coordinating motor activity from opposite sides of the body, and deficits in bimanual coordination have been documented in individuals with agenesis of or damage to the CC, particularly when the task is performed without visual feedback. The Bimanual Coordination Test was used to assess speed and accuracy on a task where both hands must coordinate to guide a cursor through angled pathways providing measures of interhemispheric interaction or the ability of the two hemispheres to coordinate activity via the corpus callosum. Twenty-one children with fetal alcohol spectrum disorders (FASD) and 17 non-exposed control children (CON), matched closely in age, sex, and ethnicity were tested. For trials with visual feedback (WV), children with FASD were slower than CON children but were equally accurate. Although statistically significant group differences were not observed on most trials completed without visual feedback (WOV), accuracy of the FASD group on WOV trials was highly variable. Group differences in accuracy on WOV angles approached significance after accounting for performance on the WV angles, and children with FASD were significantly less accurate on an individual angle believed to be particularly sensitive to interhemispheric interaction. These results indicate that children with FASD are slower than CON children but equally accurate on basic visuomotor tasks. However, as task complexity and reliance on interhemispheric interaction increases, children with FASD demonstrate variable and inaccurate performance.


Body dysmorphic disorder in a facial plastic and reconstructive surgery clinic: measuring prevalence, assessing comorbidities, and validating a feasible screening instrument.
Body dysmorphic disorder (BDD) is underrecognized and underdetected among patients undergoing facial plastic and reconstructive surgery. Patients with BDD require psychiatric care, not cosmetic surgery. To measure the prevalence of BDD in a facial plastic and reconstructive surgery clinic, to validate the Body Dysmorphic Disorder Questionnaire (BDDQ) in that setting, to assess the feasibility of implementing a standardized process for identifying patients with BDD accurately in a busy clinical practice, and to assess the comorbidity of BDD, depression, and anxiety in our patient population. We performed a prospective prevalence and instrument validation study in an academic facial plastic and reconstructive surgery clinic. We included 234 consecutive patients 18 years or older presenting to the clinic from March 3 to June 30, 2014. Prevalence of BDD as determined by the BDDQ and the criterion-standard BDD Structured Clinical Interview for DSM-IV (BDD SCID). The BDD SCID and a defect severity scale were used to confirm the BDD diagnosis and to validate the BDDQ finding. We also assessed demographic characteristics, depression, and anxiety in all patients. As confirmed by the BDD SCID, 13.1% of patients undergoing cosmetic surgery and 1.8% of those undergoing reconstructive surgery had BDD. We found the BDDQ to be an accurate (91.7%), sensitive (100%), and specific (90.3%) screening instrument for BDD. Patients with a positive BDD finding were most commonly concerned with their nose (10 of 18 [56%]), skin (5 of 18 [28%]), hair (1 of 18 [6%]), chin (1 of 18 [6%]), and ears (1 of 18 [6%]), that is, areas commonly addressed by facial plastic surgeons. Furthermore, compared with patients who did not have BDD, patients with BDD had elevated depression scores (mean [SD], 12.6 [10.8] vs 3.2 [4.2]). In the non-BDD population, mean (SD) state and trait anxiety scores resembled the norm (30.0 [7.7] and 30.3 [7.7], respectively, for men; 31.7 [10.0] and 32.1 [8.7], respectively, for women); in the BDD population, these scores were elevated (53.8 [23.4] and 50.4 [19.5], respectively, for men; 47.8 [12.7] and 49.0 [11.6], respectively, for women). Depression and anxiety scores were highly correlated in patients with BDD (r = 0.84 for patients with a BDDQ-positive screen result; r = 0.94 for patients with a positive BDD SCID finding) compared with those who did not have BDD (r = 0.51). Patients with BDD often seek cosmetic treatment. Given the documented risks and harms of surgery for patients with BDD, a systematic process for identifying such patients who seek cosmetic surgery is imperative. We found a BDD screening instrument (BDDQ) followed by the BDD SCID for patients with a BDDQ-positive screen result to be a feasible and effective way to identify patients with BDD. NA.


An investigation of the co-occurrence of panic and somatization disorders through temperamental variables.
The relation of temperamental dimensions to somatization and panic disorders in women was investigated. Eighteen patients with both panic and somatization disorders, 41 patients with panic disorder only, and 22 control subjects were compared on the Tridimensional Personality Questionnaire. The patients with somatization and panic disorders had significantly higher Novelty Seeking values than both the patients with panic only and the control subjects. The only significant correlation was between the number of symptoms of somatization disorder and Novelty Seeking. The presence of other psychiatric conditions, comorbid with panic disorder in the patient group, did not affect significantly the temperamental variables. A temperamental disposition characterized by extraversion, impulsivity, as well as frequent exploratory activity and pursuit of novel and pleasurable activities may help differentiate women with both somatization and panic disorders from women with panic disorder only.


Vitamin D in Primary Hyperparathyroidism: Effects on Clinical, Biochemical, and Densitometric Presentation.
Vitamin D (25-hydroxyvitamin D [25OHD]) deficiency (<20 ng/mL) and insufficiency (20-29 ng/mL) are common in primary hyperparathyroidism (PHPT), but data regarding their skeletal effects in PHPT are limited. The objective was to evaluate the association between 25OHD levels and PHPT severity. This is a cross-sectional analysis of 100 PHPT patients with and without 25OHD insufficiency and deficiency from a university hospital setting. We measured calciotropic hormones, bone turnover markers, and bone mineral density (BMD) by dual x-ray absorptiometry. Lower 25OHD was associated with some (PTH: r = -0.42; P < .0001; 1,25-dihydroxyvitamin D: r = -0.27; P = .008; serum PO4: r = 0.31; P = .002) but not all (serum/urine calcium) indicators of PHPT severity. Lower 25OHD was also associated with younger age, higher body mass index, male gender, better renal function, and lower vitamin D intake. Comparison of those with deficient (<20 ng/mL; 19%) vs insufficient (20-29 ng/mL; 35%) vs replete (≥30 ng/mL; 46%) 25OHD demonstrated more severe PHPT as reflected by higher PTH (mean ± SEM, 126 ± 10 vs 81 ± 7 vs 72 ± 7 pg/mL; P < .0001) but no difference in nephrolithiasis, osteoporosis, fractures, serum or urinary calcium, bone turnover markers, or BMD after adjustment for age and weight. In women, T-scores at the 1/3 radius were lower in those with 25OHD of 20-29 ng/mL, compared to those who were vitamin D replete (P = .048). In multiple regression modeling, 25OHD (but not PTH) was an independent predictor of 1/3 radius BMD. Vitamin D deficiency is associated with more severe PHPT as reflected by PTH levels, but effects on BMD are limited to the cortical 1/3 radius and are quite modest. These data support international guidelines that consider PHPT patients with 25OHD <20 ng/mL to be deficient. However, in this cohort with few profoundly vitamin D-deficient patients, vitamin D status did not appear to significantly impact clinical presentation or bone density.


Popliteal artery aneurysms: is endovascular reconstruction durable?
To describe an endovascular method of performing femoropopliteal in situ saphenous vein (SV) bypass and popliteal artery aneurysm (PAA) embolization. Twenty-two patients underwent PAA operations. Twelve patients had conventional SV bypasses with PAA proximal and distal ligation, whereas 10 underwent PAA embolization and an endovascular in situ SV bypass (EISB). The endovascular procedure was performed using an angioscopically guided side branch coil occlusion system. The PAAs were coil embolized under fluoroscopic surveillance. No deaths or wound complications occurred in the EISB group. The mean hospital length of stay (LOS) was 2.1 days. Seven EISB procedures were performed through 2 incisions, whereas 3 operations required an additional incision. One graft occluded at 3 months. All PAAs remained occluded by color-flow ultrasonography at follow-up ranging from 4 to 23 months (mean 13.6); cumulative primary patency was 89%. In the conventional bypass group, no deaths occurred, but 3 (25%) patients had wound complications. The mean LOS was 6.2 days, and 1 graft occluded, giving an 86% cumulative primary patency at 42 months. This minimally invasive technique obviates an extensive incision to harvest the SV and ligate the PAA proximally and distally. If long-term endovascular bypass graft patency and PAA occlusion rates prove to be similar to open operative results, the benefits of reduced wound complications, decreased hospital LOS, and increased health care savings support further investigation of this endovascular approach for the treatment of PAA.


A summertime peak of "winter vomiting disease": surveillance of noroviruses in England and Wales, 1995 to 2002.
Noroviruses are the most common cause of gastroenteritis outbreaks in industrialised countries. Gastroenteritis caused by Norovirus infection has been described as a highly seasonal syndrome, often referred to as "winter vomiting disease". The Public Health Laboratory Service Communicable Disease Surveillance Centre has systematically collected reports of laboratory confirmed cases of Norovirus-gastroenteritis since 1995. We analysed these data for annual and seasonal trends and age distribution. A mid-summer peak in reported cases of Norovirus was observed in 2002, unlike all six previous years when there was a marked summer decline. Total reports from 2002 have also been higher than all previous years. From the first 10 months of 2002, a total of 3029 Norovirus diagnoses were reported compared the previous peak in 1996 of 2437 diagnoses for the whole 12-month period. The increase in 2002 was most marked in the 65 and older age group. This surveillance data challenges the view that Noroviruses infections exclusively have wintertime seasonality.


Presence and activity of compounds with GnRH-like activity in the ovary of seabream Sparus aurata.
The binding and activity of gonadotropin-releasing hormone (GnRH) were characterized in the mature gilthead seabream (Sparus aurata) ovary by use of an analogue of salmon GnRH([D-Arg6,Trp7,Leu8,Pro9-N(Et)]GnRH, sGnRH-A) as labeled ligand. The binding of 125I-sGnRH-A to the seabream ovarian membrane preparation was saturable, displaceable, reversible, and dependent on time, temperature and tissue concentration. Addition of unlabeled s-GnRH-A displaced the radio-ligand in a dose-related manner, indicating the presence of one class of high-affinity binding sites with an equilibrium dissociation constant of 45.5 +/- 6.2 nM. Addition of other GnRH peptides, including salmon GnRH ([Trp7,Leu8]GnRH) and chicken GnRH-II ([His5,Trp7,Tyr8]GnRH), also displaced 125I-sGnRH-A; all these peptides bound with lower affinities than sGnRH-A to the seabream ovarian binding site. In this study, we also demonstrated the presence of compounds with GnRH-like activity in the ovary of seabream. Seabream ovarian extract stimulated pituitary gonadotropin release from the goldfish pituitary and displaced 125I-sGnRH-A binding in the seabream ovary. Furthermore, addition of sGnRH-A to cultured seabream oocytes directly stimulated reinitiation of oocyte meiosis, as indicated by germinal vesicle breakdown. Overall, the present study characterizes GnRH-binding sites in the seabream ovary and supports the hypothesis that GnRH or compounds with GnRH-like activity play an autocrine/paracrine role in the regulation of ovarian function in the seabream ovary.


Sex differences in symptom presentation and their impact on diagnostic accuracy in Werner syndrome.
Whether sex differences exist in hereditary progeroid syndromes remains unclear. In this study, we investigated sex differences in patients with Werner syndrome (WS), a model of human aging, using patient data at the time of diagnosis. The presence of six cardinal signs in the diagnostic criteria was retrospectively evaluated. We found that the percentage of patients with all cardinal signs was higher in males than in females (54.2% vs. 21.2%). By the age of 40 years, 57.1% of male patients with WS presented with all the cardinal signs, whereas none of the female patients developed all of them. In particular, the frequency of having a high-pitched, hoarse voice, a characteristic of WS, was lower in female patients. The positive and negative predictive values for clinical diagnosis were 100% for males and females, indicating the helpfulness of diagnostic criteria regardless of sex. More female patients than male (86.7% vs. 64%) required genetic testing for their diagnosis because their clinical symptoms were insufficient, suggesting the importance of genetic testing for females even if they do not show typical symptoms of WS. Finally, the frequency of abnormal voice was lower in patients with WS harboring the c.3139-1G > C homozygous mutation. These results indicate, for the first time, that there are sex differences in the phenotypes of hereditary progeroid syndromes. The analysis of this mechanism in this human model of aging may lead to the elucidation of sex differences in the various symptoms of normal human aging. Geriatr Gerontol Int 2023; ••: ••-••.


Short- vs. long-circulating magnetoliposomes as bone marrow-seeking MR contrast agents.
We evaluated the relaxation enhancement and biodistribution of short- vs. long-circulating magnetoliposomes as a new contrast agent for magnetic resonance (MR) imaging of bone marrow. Magnetoliposomes with (ML-PEG) and without (ML) incorporation of polyethylene glycol (PEG, Mw 2000) were prepared, measuring 40 nm in diameter with 1-6 iron oxide crystals/vesicle. PEGylation selectively enhanced the T2 relaxivity of magnetoliposomes by 10% to 15%, with R1 and R2 values of 3 and 240 s(-1)/mM at 1.5 T and 37 degrees C. ML (n = 6) and ML-PEG (n = 6) preparations were administered IV into young (6-8 weeks old) and adult (>1 year old) Sprague-Dawley rats at 100 pmol Fe/kg. PEGylation increased blood half-life (P<0.05 for t > 30 minutes), following a biexponential clearance with a long half-life of 53.2+/-13.2 minutes. The clearance of ML was monoexponential, with a half-life 7.4+/-0.4 minutes. MR imaging revealed a pronounced uptake in bone marrow, including the iliac bone, femur, tibia, and upper and lower vertebrae. The bone marrow uptake of ML-PEG was comparable to that of ML, with both reaching a plateau within 30 minutes following injection. Fast spin-echo T2-weighted imaging was found to provide optimal contrast enhancement and allowed a clear depiction of red to yellow marrow conversion due to normal aging. While the use of magnetoliposomes can provide the added benefit of therapeutic drug or gene delivery, further investigation is warranted to assess their usefulness in differentiating normal vs. abnormal marrow conditions.


A potential risk assessment of a dengue outbreak in north central Texas, USA. (Part 1 of 2): Abundance and temporal variation of dengue vectors.
In response to three imported dengue cases in north central Texas as well as increased case numbers in Texas and adjoining Mexican states in 2005, the authors assessed the potential risk of a dengue outbreak in north central Texas by investigating abundance and temporal variation of dengue vectors in 2006. Dengue vector abundance was monitored from 54 sites in Dallas County, Texas, from June to November 2006, using oviposition traps. Both dengue vectors--the yellow fever mosquito, Aedes aegypti, and the Asian tiger mosquito, Aedes albopictus--were present. Of the two, Ae. albopictus was more abundant and its abundance appeared to be positively affected by temperature and precipitation. Potential risk of a dengue outbreak was predicted based on the abundance and temporal variation of dengue vectors and a long-term trend of breeding season precipitation and warmer winter temperatures.


Comparing three different methods to detect selective loci using dominant markers.
We carried out a simulation study to compare the efficiency of three alternative programs (DFDIST, DETSELD and BAYESCAN) to detect loci under directional selection from genome-wide scans using dominant markers. We also evaluated the efficiency of correcting for multiple testing those methods that use a classical probability approach. Under a wide range of scenarios, we conclude that BAYESCAN appears to be more efficient than the other methods, detecting a usually high percentage of true selective loci as well as less than 1% of outliers (false positives) under a fully neutral model. In addition, the percentage of outliers detected by this software is always correlated with the true percentage of selective loci in the genome. Our results show, nevertheless, that false positives are common even with a combination of methods and multitest correction, suggesting that conclusions obtained from this approach should be taken with extreme caution.


Sequence of central nervous system myelination in human infancy. I. An autopsy study of myelination.
This study establishes the sequence of myelination in a population of autopsied infants from birth through the second postnatal year. Myelination was assessed in 62 precisely defined central nervous system (CNS) sites of 162 infants with diverse diseases who were autopsied from 1972 to 1984 at Children's Hospital, Boston. The degree of myelination was graded on an ordinal scale of 0-4 using the inferior cerebellar peduncle as an internal standard. This grading system is a modification of that used for fetal myelination in the National Collaborative Perinatal Project (NCPP). The data are summarized by, median degree of myelination for each age group and site; and Ayer estimates for the age at which at least 10, 50, and 90% of infants reach a particular myelin degree in each site. "Marker" sites in the cerebrum are provided for the pathologist to compare myelination between an individual infant brain and the brains from our autopsy population. These data should be useful in identifying diverse peri- and postnatal conditions affecting myelination in human infancy. They also provide guidelines for the assessment of CNS myelination by sophisticated imaging techniques in living infants.


Hsp90 regulates the phosphorylation and activity of serum- and glucocorticoid-regulated kinase-1.
SGK-1 (serum- and glucocorticoid-regulated kinase-1), a member of the AGC protein kinase family, plays an important role in regulating ion channel expression and contributes to malignant epithelial cell proliferation and survival. SGK-1 activity is regulated on three levels: transcriptional induction following a variety of environmental and intracellular stresses, proteasomal degradation, and phosphorylation. Here we report that phosphoinositide 3-kinase (PI3K)-dependent phosphorylation of SGK-1 requires formation of a complex between SGK-1 and heat-shock protein 90 (Hsp90). Inactivation of Hsp90 by geldanamycin led to decreased SGK-1 phosphorylation independently of increased proteasomal protein degradation, and inhibition of PI3K activity by LY294002 appeared to eliminate SGK-1 phosphorylation at the same residues as those affected by geldanamycin treatment. Interestingly, geldanamycin-targeted phosphorylation sites were not limited to the known conserved PI3K-dependent sites Thr-256 and Ser-422 in SGK-1 but included additional unknown PI3K-dependent residues. Inhibition of Hsp90 also resulted in a complete loss of SGK-1 kinase activity, suggesting that Hsp90 activity is essential for regulating the PI3K/SGK-1 pathway.


Microscopic interactions of the imidazolium-based ionic liquid with molecular liquids depending on their electron-donicity.
Microscopic interactions of an imidazolium-based ionic liquid, 1-ethyl-3-methylimidazolium bis(trifluoromethanesulfonyl)imide (C2mimTFSI), with dimethyl sulfoxide (DMSO), methanol (MeOH), and acetonitrile (AN) have been analyzed by means of Raman, attenuated total reflectance infrared (ATR-IR), (1)H and (13)C NMR spectroscopy techniques. The magnitude of the red-shift of the C(2)-H vibration mode of the imidazolium ring and the deshielding of the C(2)-H hydrogen and carbon atoms, compared with that of the other atoms of the ring or the anion, indicated a strong interaction between the C(2)-H hydrogen atom and the molecular liquids in the following order; DMSO ≫ MeOH > AN. This correlates with the order of the electron donicities of these molecular liquids which allows us to suggest a hydrogen bonding character of these interactions. The behavior of S= O vibration of DMSO as a function of the DMSO molar fraction xDMSO also suggested that DMSO molecules are stoichiometrically hydrogen-bonded with the three hydrogen atoms, C(2,4,5)-H, of the ring. In contrast, the hydrogen bonding between MeOH and the C(4,5)-H atoms is much weaker than that in DMSO. AN hardly forms hydrogen bonds with the C(4,5)-H atoms. Instead, AN molecules may interact with the imidazolium ring through the π-π interaction. The interactions between the imidazolium ring and the molecular liquids lead to the loosening of the TFSI anion from the cation; this correlates with both the blue-shift of the S=O stretching vibration of TFSI and the deshielding of the trifluoromethyl carbon atoms with an increase in the molar fraction of the molecular liquid xML. The latter is weak in the MeOH solutions, and may be explained by the possible hydrogen bonding of the MeOH hydroxyl group as an electron-acceptor with the TFSI anion. Furthermore, the organization of MeOH molecules around the ethyl and methyl groups of the cation is discussed in terms of the chemical shift of the hydrogen and carbon atoms in these groups as a function of xML.


Transactivation of a ribosomal gene by simian virus 40 large-T antigen requires at least three activities of the protein.
Simian virus 40 large-T antigen transactivates the ribosomal genes which are transcribed by RNA polymerase (pol I), as well as genes that are dependent on either pol II or pol III. This report identifies regions and activities of T antigen that are required to transactivate a pol I-dependent rat ribosomal gene promoter. By using the rat ribosomal gene (rDNA) promoter linked to a chloramphenicol acetyltransferase gene, we show that at least three separable T-antigen regions are necessary to achieve wild-type levels of transactivation of rDNA in transiently transfected monkey cells. One activity depends on the region of T antigen shared with small-t antigen (T/t common region). A second activity maps to amino acids 109 to 626 and is highly sensitive to mutational inactivation. Complementation analyses suggest that at least one activity in this region is independent of and must be in cis with the activity within the T/t common region. In addition, a functional nuclear localization signal is required for maximal T-antigen-mediated transactivation of rat rDNA. The three activities work in concert to override cellular species-specific controls and transactivate the rat ribosomal gene promoter. Finally, we provide evidence that although the tumor suppressor protein Rb has been shown to repress a pol I-dependent promoter, transactivation of the rat rDNA promoter does not depend on T antigen's ability to bind the tumor suppressor product Rb.


Protein AA amyloidosis in Nigeria.
Eight cases of multi-system amyloid have been found in 4235 autopsies (0.2%) performed at University College Hospital, Ibadan, Nigeria between 1970 and 1979. Blocks and sections of seven cases showed the organ distribution associated with secondary amyloid, involving kidney, spleen and liver. Potassium permanganate oxidation prior to Congo Red staining showed that all seven cases contained protein AA amyloid. Five patients had tuberculosis but in two patients there was no recognized cause, idiopathic AA amyloid. Patients usually presented with nephrotic syndrome or renal failure and hypertension was absent. There appears to be a low incidence of amyloidosis in Africans, despite the occurrence of a large number of potential amyloidogenic stimuli, indicating the importance of individual host reactivity and failure of amyloid degradation in the causation of amyloidosis.


Minimum prick test panel for adult patients with asthma and rhinitis in Ankara, Turkey.
Determination of the number and type of allergens needed to be tested in epidemiological studies is important in order to identify most of the sensitized subjects with a cost-effective approach. This study aimed to investigate the minimum skin prick test panel for the identification of at least 95% of the sensitized subjects with symptoms of asthma and/or allergic rhinitis (AR) in Ankara, Turkey. Skin prick test results of 7492 patients who were referred to our outpatient clinic with clinical symptoms of asthma and/or AR between 1991 and 2005 were evaluated retrospectively. Seven allergens were tested in all and 13 allergens in 4202 patients. The allergen group needed for detection of 95% of the sensitized subjects was determined for both the 7 and 13 allergen panels. The study protocol was approved by the local ethics committee of Hacettepe University. The atopy prevalences in the whole study population and in 4202 patients tested with the 13 allergen panel were calculated as 32.2% and 42.6%, respectively. Three allergens (Phleum pratense, Dermatophagoides pteronyssinus and Artemisia vulgaris) within the 7 allergen panel were adequate for the identification of at least 95% of the sensitized subjects. Olea europae was added to the previous three allergens when the 13 allergen panel was applied. Three to four allergens are sufficient for identification at least 95% of sensitized subjects with asthma and/or AR in Ankara, Turkey.


How to Test Human CAR T Cells in Solid Tumors, the Next Frontier of CAR T Cell Therapy.
Chimeric antigen receptor (CAR) T cell therapy has proven to be a successful treatment option for leukemias and lymphomas. These encouraging outcomes underscore the potential of adoptive cell therapy for other oncology applications, namely, solid tumors. However, CAR T cells are yet to succeed in treating solid tumors. Unlike liquid tumors, solid tumors create a hostile tumor microenvironment (TME). CAR T cells must traffic to the TME, survive, and retain their function to eradicate the tumor. Nevertheless, there is no universal preclinical model to systematically test candidate CARs and CAR targets for their capacity to infiltrate and eliminate human solid tumors in vivo. Here, we provide a detailed protocol to evaluate human CAR CD4+ helper T cells and CD8+ cytotoxic T cells in immunodeficient (NSG) mice bearing antigen-expressing human solid tumors.


IL-15 induces the expression of chemokines and their receptors in T lymphocytes.
IL-15 is a T cell growth factor that shares many biological activities with IL-2 and uses the same beta/gamma polypeptides of the IL-2R complex for signal transduction. Accumulating evidence implicates an important role for this cytokine in the inflammatory response of the host. Consistent with such a role, IL-15 has been shown to be a chemoattractant for T lymphocytes, NK cells, and neutrophils. Extending these observations, we now show that IL-15 is a potent inducer of CC-, CXC-, and C-type chemokines in T lymphocytes. In addition, we demonstrate that IL-15 induces CC chemokine receptors, but not CXC chemokine receptors, in a dose-dependent manner. Thus, our findings suggest that the proinflammatory effects of IL-15 at least in part may be due to the induction of chemokines and their receptors in T cells. Furthermore, we demonstrate that IL-15 promotes entry and replication of macrophage-tropic HIV in T lymphocytes and suggest a plausible mechanism by which IL-15, a cytokine that is elevated in HIV-infected individuals, may promote the transition of HIV displaying the M-tropic phenotype primarily associated with the initial transmission into the T cell-tropic phenotype that predominates as the disease progresses.


Routine EGFR and KRAS Mutation analysis using COLD-PCR in non-small cell lung cancer.
Aims: Epidermal growth factor receptor (EGFR) antagonists are particularly active in non-small cell lung cancer (NSCLC) patients with tumours bearing mutations in the EFGR gene. EGFR mutation prevalence is very low in squamous histology. Response rates using these drugs in patients with KRAS mutations are low, so available KRAS mutation information may aid treatment selection in the second-line setting. Since 2009, patients presenting to this hospital with non-squamous histology have been routinely screened for mutations in both the EGFR and KRAS genes, with results used to inform treatment. We present an analysis of 215 consecutive patients for whom EGFR mutation analysis was informative. Methodology: EGFR and KRAS mutations were identified using a COLD-PCR technique confirmed with sequencing, which makes no prior assumption about location of specific mutations. Results were correlated with clinical and demographic data from hospital records, where available. Results: The prevalence of patients with EGFR mutations was 14% and for KRAS mutations it was 27%. Despite the conventional understanding that EGFR and KRAS mutations are mutually exclusive, we identified two dual mutations. Of 29 patients identified with mutated EGFR, there were 3/8/8/10 mutations in exons 18/19/20/21 respectively. Exon 20 mutations were identified in a proportion exceeding many other series because of the unbiased mutation analysis used, and clinical benefit was seen in some of these. Of 23 different EGFR mutations identified, 11 have not previously been described in the literature. Conclusions: The high prevalence of EGFR, KRAS or both mutations (40%) in this non-squamous population tested in clinical practice supports a policy of routine screening for these mutations in NSCLC.


Levodopa-responsive parkinsonism in a patient with corticobasal degeneration and bilateral choroid plexus xanthogranulomas.
Corticobasal degeneration (CBD) has substantial overlap of clinical features with other neurodegenerative diseases including Parkinson's disease (PD). Its clinical diagnostic accuracy is the lowest among the common neurodegenerative diseases, and its antemortem diagnosis is more challenging when CBD is comorbid with another brain disease. We report an elderly male patient with multiple medical conditions and a family history of essential tremor. He presented with progressive tremor that was initially thought to be essential tremor and later diagnosed as PD despite head computerized tomography showing bilateral intraventricular masses and other minor changes. The clinical diagnosis of PD was supported by his responsiveness to low-dose levodopa. However, postmortem neuropathological examination revealed CBD and bilateral choroid plexus xanthogranulomas with mild ventricular enlargement and multifocal ependymal lining injury presumably due to mild hydrocephalus. CBD is typically levodopa-unresponsive, but hydrocephalus-associated parkinsonism is commonly levodopa-responsive. We raise awareness of the present comorbidity and atypical parkinsonism due to the choroid plexus xanthogranuloma-induced hydrocephalus for the clinical diagnosis and management of parkinsonism.


Laryngeal involvement by thyroid disease.
Thyroid tissue or neoplastic derivatives therefrom in the larynx or trachea have their genesis in either ectopia or a direct invasion by carcinoma of the thyroid gland proper. The benign lesions are found preponderantly in patients from regions in which goiter is endemic, and they may represent as much as 7% of all benign tracheal tumors. Laryngotracheal invasion by thyroid carcinoma also has a frequency of approximately 7%, with all histologic types represented.


A point mutation affecting an SP1 binding site in the promoter of the ferrochelatase gene impairs gene transcription and causes erythropoietic protoporphyria.
Clinical manifestation of erythropoietic protoporphyria (EPP) results from coinheritance of a mutated allele and a wild-type low-expressed allele of the ferrochelatase (FECH) gene. Currently, up to 90 different mutations affecting the coding region or splicing junctions of the FECH gene have been identified. Despite the high molecular heterogeneity, no functional mutations have been previously reported in the promoter region. The weaker allele expression has been controversially associated to the presence of different intragenic polymorphisms. We applied a two-step screening strategy using denaturing gradient gel electrophoresis followed by direct sequencing in order to rapidly identify FECH gene mutations in Italian EPP patients. We identified two unrelated subjects showing a normal FECH coding region but a single G>C base substitution at position -250 in the FECH promoter and the -251G, IVS1-23T, and IVS3-48C polymorphisms in trans to the substitution. To investigate the effect of the -250G>C mutation on protein binding to the FECH promoter, we conducted electro mobility shift assay (EMSA) and supershift analysis. To determine its effect on the transcriptional activity, K562 and Jurkat cell lines were transiently transfected. EMSA showed that the -250G>C mutation results in the loss of an SP1 binding site, and transient transfection assays demonstrated that such mutation strongly impairs promoter activity. Moreover, we showed that the -251A>G polymorphism, although unable to affect SP1 binding, displays a significant reduction in the transcriptional activity of the promoter. This is the first report of a mutation in the FECH promoter affecting binding of a transcription factor and causing EPP phenotype.


[Study of the relation between transformational leadership of the head nurse and supplementary efforts of nursing personnel].
The present study proposes to show that the ethos of the Head Nursing staff is related to the practice of transformational leadership, and to additional efforts displayed by the treatment staff. Using a sampling of 57 nurses, it was established that transformational leadership presented a positive influence on additional efforts which were superior to that produced by transactional leadership. Contrary to transactional leadership, transformational leadership was shown to exert a positive effect on the ethos of the leader who in turn, had a positive influence on the extra care supplied by the nursing staff.


Job satisfaction of Malaysian registered nurses: a qualitative study.
Job satisfaction is an important factor in health care settings. Strong empirical evidence supports a causal relationship between job satisfaction, patient safety and quality of care. However, there have not been any studies exploring the job satisfaction of Malaysian nurses. The main purpose of this qualitative descriptive study was to explore the factors related to feelings of job satisfaction as well as job dissatisfaction experienced by registered nurses in Malaysia. A convenient sample of 46 Malaysian nurses recruited from a large hospital (number of beds = 895) participated in the study. A total of seven focus group discussions were conducted with nurses from surgical, medical and critical care wards. A semi-structured interview guide was used to facilitate the interviews, which were audio-recorded, transcribed verbatim and checked. The transcripts were used as data and were analysed using a thematic approach. The study identified three main themes that influenced job satisfaction: (1) nurses' personal values and beliefs; (2) work environment factors and (3) motivation factors. Concerning the nurses' personal values and beliefs, the ability to help people made the nurses felt honoured and happy, which indirectly contributed to job satisfaction. For work environment factors, team cohesion, benefit and reward, working conditions play an important role in the nurses' job satisfaction. Motivation factors, namely, professional development and clinical autonomy contributed to job satisfaction. It is important for nurse leaders to provide more rewards, comfortable work environments and to understand issues that affect nurses' job satisfaction. Our findings highlight the importance of factors that can improve nurses' job satisfaction. The study provides basic information for hospital administrators in planning effective and efficient policies to improve nursing job satisfaction in order to increase the quality of patient care and decrease nursing turnover.


Parental care buffers against inbreeding depression in burying beetles.
When relatives mate, their inbred offspring often suffer a reduction in fitness-related traits known as "inbreeding depression." There is mounting evidence that inbreeding depression can be exacerbated by environmental stresses such as starvation, predation, parasitism, and competition. Parental care may play an important role as a buffer against inbreeding depression in the offspring by alleviating these environmental stresses. Here, we examine the effect of parental care on the fitness costs of inbreeding in the burying beetle Nicrophorus vespilloides, an insect with facultative parental care. We used a 2 × 2 factorial design with the following factors: (i) the presence or absence of a caring female parent during larval development and (ii) inbred or outbred offspring. We examined the joint influence of maternal care and inbreeding status on fitness-related offspring traits to test the hypothesis that maternal care improves the performance of inbred offspring more than that of outbred offspring. Indeed, the female's presence led to a higher increase in larval survival in inbred than in outbred broods. Receiving care at the larval stage also increased the lifespan of inbred but not outbred adults, suggesting that the beneficial buffering effects of maternal care can persist long after the offspring have become independent. Our results show that parental care has the potential to moderate the severity of inbreeding depression, which in turn may favor inbreeding tolerance and influence the evolution of mating systems and other inbreeding-avoidance mechanisms.


Diffusion-tensor MR imaging in children with developmental delay: preliminary findings.
To determine whether diffusion-tensor magnetic resonance (MR) imaging can depict abnormalities in patients with a diagnosis of developmental delay but structurally normal brain MR imaging results. Twenty pediatric patients who received a diagnosis of developmental delay underwent brain MR examinations, including diffusion-tensor MR imaging. The MR findings in these patients were compared with those in 10 age-matched neurodevelopmentally healthy children. Diffusion constant (Dav) and anisotropy were measured bilaterally in regions of interest in the centrum semiovale, corona radiata, internal capsule, corpus callosum, and subcortical white matter of the frontal and parieto-occipital lobes. By using a one-tailed Student t test in the positive direction for Dav and in the negative direction for anisotropy and P <.05 to indicate a significant difference, the Dav and anisotropy values for children with developmental delay were compared with those for children who were neurodevelopmentally healthy. The children with developmental delay had significant increases in Dav in all measured structures (P, <.001 to <.03). Significant decreases in anisotropy were detected in all white matter fiber tracts studied (P, <.001 to <.03) except the posterior limb of the internal capsule. In the children with developmental delay, diffusion-tensor MR imaging depicted decreases in anisotropy and increases in Dav in the white matter fiber tracts, which appeared to be normal at conventional MR imaging.


Variation in Pyrethroid Resistance Phenotypes in Anopheles darlingi in an Area with Residual Malaria Transmission: Warning of Suspected Resistance in French Guiana.
Anopheles darlingi is the main vector of malaria in South America. In French Guiana, malaria transmission occurs inland and along the rivers with a regular reemergence in the lower Oyapock area. Control against malaria vectors includes indoor residual spraying of deltamethrin and the distribution of long-lasting impregnated bednets. In this context, the level of resistance to pyrethroids was monitored for 4 years using CDC bottle tests in An. darlingi populations. A loss of susceptibility to pyrethroids was recorded with 30-minute knock-down measured as low as 81%. However, no pyrethroid molecular resistance was found by sequencing a 170 base pair fragment of the S6 segment of domain II of the voltage-gated sodium channel gene. Fluctuation of resistance phenotypes may be influenced by the reintroduction of susceptible alleles from sylvatic populations or by other mechanisms of metabolic resistance.


Prediction of nutrient digestibility in grower-finisher pigs based on faecal microbiota composition.
Microbiota play an important role in total tract nutrient digestion, especially when fibrous diets are fed to pigs. This study aimed to use metagenomics to predict faecal nutrient digestibility in grower-finisher pigs. The study design consisted of 160 three-way crossbreed grower-finisher pigs (80 female and 80 male) which were either fed a diet based on corn/soybean meal or a more fibrous diet based on wheat/barley/by-products. On the day before slaughter, faecal samples were collected and used to determine faecal digestibility of dry matter, ash, organic matter, crude protein, crude fat, crude fibre and non-starch polysaccharides. The faecal samples were also sequenced for the 16S hypervariable region of bacteria (V3/V4) to profile the faecal microbiome. With these data, we calculated the between-animal variation in faecal nutrient digestibility associated with variation in the faecal microbiome, that is the "microbiability". The microbiability values were significantly greater than zero for dry matter, organic matter, crude protein, crude fibre and non-starch polysaccharides, ranging from 0.58 to 0.93, as well as for crude fat with a value of 0.37, but not significantly different from zero for ash. Using leave-one-out cross-validation, we estimated the accuracy of predicting digestibility values of individual pigs based on their faecal microbiota composition. The accuracies of prediction for crude fat and ash digestibility were virtually 0, and for the other nutrients, the accuracies ranged from 0.42 to 0.63. In conclusion, the faecal microbiota composition gave high microbiability values for faecal digestibility of dry matter, organic matter, crude protein, crude fibre and non-starch polysaccharides. The accuracies of prediction are relatively low if the interest is in precisely predicting faecal nutrient digestibility of individual pigs, but are promising from the perspective of ranking animals in a genetic selection context.


Chitosans as absorption enhancers for poorly absorbable drugs. 1: Influence of molecular weight and degree of acetylation on drug transport across human intestinal epithelial (Caco-2) cells.
Chitosan has recently been demonstrated to effectively enhance the absorption of hydrophilic drugs such as peptides and proteins across nasal and intestinal epithelia (1-3). In this study, the effect of the chemical composition and molecular weight of chitosans on epithelial permeability and toxicity was investigated using monolayers of human intestinal epithelial Caco-2 cells as a model epithelium. Eight chitosans varying in degree of acetylation (DA) and molecular weight were studied. The incompletely absorbed hydrophilic marker molecule 14C-mannitol was used as a model drug to assess absorption enhancement. Changes in intracellular dehydrogenase activity and cellular morphology were used to assess toxicity. Chitosans with a low DA (1 and 15%) were active as absorption enhancers at low and high molecular weights. However, these chitosans displayed a clear dose-dependent toxicity. Chitosans with DAs of 35 and 49% enhanced the transport of 14C-mannitol at high molecular weights only, with low toxicity. One chitosan (DA = 35%; MW = 170 kD) was found to have especially advantageous properties such as an early onset of action, very low toxicity, and a flat dose-absorption enhancement response relationship. The structural features of chitosans determining absorption enhancement are not correlated with those determining toxicity, which makes it possible to select chitosans with maximal effect on absorption and minimal toxicity.


Prevalence of C-shaped Configurations in the Mandibular First and Second Premolars: A Cone-beam Computed Tomographic In Vivo Study.
The mandibular premolar C-shaped anatomy has been presented as a complex morphology to be treated. The objective of this retrospective observational study was to evaluate and characterize, in in vivo conditions, the incidence of these morphologies using cone-beam computed tomographic (CBCT) technology. Mandibular premolar CBCT samples were collected from a preexisting database. All teeth were analyzed in 3 planes (axial, coronal, and sagittal), and the C-shape classification was performed at 3 different axial levels (coronal, middle, and apical). C-shape presence and configuration were recorded as well as the number of roots, the presence of a radicular groove, and Vertucci classification. The Z test for proportions was used to analyze the differences between independent groups. Intraobserver reliability was tested using the Cohen kappa test. Two thousand twelve mandibular premolars were included in this study. A prevalence of C-shaped morphologies was noted in 2.3% and 0.6% of mandibular first and second premolars, respectively. This clinical condition was mostly unilateral. The C-shaped configuration (C1 and C2) was found mainly in the middle axial level. Its presence was uncommon in the apical level and null in the coronal level; 61.5% of all mandibular first premolar C shapes were identified in Vertucci type V roots. Differences were observed among sex, teeth, and Vertucci root configuration at P < .05. Although the mandibular premolar C-shaped anatomy has a low prevalence ratio, a clinician should be aware of its existence, mainly when treating roots with Vertucci type V configuration. The prevalence was statistically higher in first premolars and males.


Changes in gene expression during chemical-induced nephrocarcinogenicity in the Eker rat.
Hydroquinone (HQ) is a rodent carcinogen and a potential human carcinogen. Glutathione conjugation of HQ enhances its biological reactivity, and 2,3,5-tris-(glutathion-S-yl)hydroquinone (TGHQ) is a potent nephrotoxicant and nephrocarcinogen in the Eker rat. Moreover, a single exposure of primary epithelial cells derived from Eker rat kidneys to TGHQ transforms these cells into an immortalized phenotype (quinol-thioether transformed rat renal epithelial (QT-RRE) cells). The Eker rat bears a mutation in one allele of the tuberous sclerosis-2 (Tsc-2) tumor suppressor gene, which predisposes the animals to the development of spontaneous and chemical-induced renal cell carcinoma. Thus, the Eker rat provides a unique model for elucidating the mechanisms of renal tubular epithelial carcinogeneisis. cDNA microarray analysis of QT-RRE3 cells and of tumor tissue derived from the kidneys of Eker rats treated with TGHQ revealed alterations (by threefold or greater) in the expression of a total of 80 genes. Fifteen percent of these genes exhibited similar expression patterns in both QT-RRE cells and tumor tissue. The differentially expressed genes primarily participate in three major areas: (1) signal transduction or in the regulation of signal transduction (extracellular signal regulated kinase 2 (ERK2); protein kinase CK2; protein kinase B; c-jun; NF-kappaB; ras-related GTPases; annexins), (2) stress response, tissue remodeling, and DNA repair (glutathione-S-transferases; procollagen c proteinase enhancer; plasminogen activator; tissue inhibitor of metalloprotease 3; apurinic/apyrimidic endonuclease), and (3) electron transport and energy homeostasis (cytochrome c oxidase subunits). The changes in the expression of many of these genes was confirmed by reverse transcription (RT)-polymerase chain reactions (PCR) using primers specific for the differentially expressed genes. As an example, the annexin I and II genes, implicated in signal transduction, were highly induced in tumor tissue and also in dysplastic lesions isolated from the kidneys of rats treated chronically with TGHQ. The annexin I and II proteins were also upregulated in tumor tissue, which probably play an important role in TGHQ-induced nephrocarcinogenesis. Moreover, in the present study, a tumorigenicity assay using athymic nude mice revealed that QT-RRE cell lines formed tumors when injected in the subcutis of nude mice, providing evidence that the cells are malignantly transformed. Histopathological analysis further indicated that the tumors were composed of neoplastic cells, resembling renal carcinoma cells with varying degrees of atypia, with the presence of apoptotic and mitotic figures.


Mini-guts in a dish: Perspectives of adult Cystic Fibrosis (CF) patients and parents of young CF patients on organoid technology.
Organoid technology enables the cultivation of human tissues in a dish. Its precision medicine potential could revolutionize the Cystic Fibrosis (CF) field. We provide a first thematic exploration of the patient perspective on organoid technology to set the further research agenda, which is necessary for responsible development of this ethically challenging technology. 23 semi-structured qualitative interviews with 14 Dutch adult CF patients and 12 parents of young CF patients to examine their experiences, opinions, and attitudes regarding organoid technology. Four themes emerged: (1) Respondents express a close as well as a distant relationship to organoids; (2) the open-endedness of organoid technology sparks hopes and concerns, (3) commercial use evokes cautiousness. (4) Respondents mention the importance of sound consent procedures, long-term patient engagement, responsible stewardship, and stringent conditions for commercial use. The precision medicine potential of organoid technology can only be realized if the patient perspective is taken adequately into account.


A twenty eight-day repeated dose toxicity study of black soybean extract in Sprague-Dawley rats.
This study was designed to evaluate any adverse effect of a hot water extract of black soybeans (Glycine max (L.) Merr.), when administered to both sexes of Crj:CD(SD)IGS rats at dietary levels of 0 (control), 0.5, 1.5 and 5.0% (6 rats/sex/group). During the study, the treatment had no adverse effects on clinical signs, survival, body weights, and food and water consumption, or on findings of ophthalmology, urinalysis, hematology, or blood biochemistry. Organ weights, gross pathology and histopathology exhibited no differences of toxicological significance between control and treated rats. Thus, the no-observed-adverse-effect level (NOAEL) of black soybean extract was concluded to be 5.0% (3,618 mg/kg body weight/day for males and 4,066 mg/kg body weight/day for females) from the present study.


Development and performance of a sleep estimation algorithm using a single accelerometer placed on the thigh: an evaluation against polysomnography.
Accelerometers placed on the thigh provide accurate measures of daily physical activity types, postures and sedentary behaviours, over 24 h and across consecutive days. However, the ability to estimate sleep duration or quality from thigh-worn accelerometers is uncertain and has not been evaluated in comparison with the 'gold-standard' measurement of sleep polysomnography. This study aimed to develop an algorithm for sleep estimation using the raw data from a thigh-worn accelerometer and to evaluate it in comparison with polysomnography. The algorithm was developed and optimised on a dataset consisting of 23 single-night polysomnography recordings, collected in a laboratory, from 15 asymptomatic adults. This optimised algorithm was then applied to a separate evaluation dataset, in which, 71 adult males (mean [SD] age 57 [11] years, height 181 [6] cm, weight 82 [13] kg) wore ambulatory polysomnography equipment and a thigh-worn accelerometer, simultaneously, whilst sleeping at home. Compared with polysomnography, the algorithm had a sensitivity of 0.84 and a specificity of 0.55 when estimating sleep periods. Sleep intervals were underestimated by 21 min (130 min, Limits of Agreement Range [LoAR]). Total sleep time was underestimated by 32 min (233 min LoAR). Our results evaluate the performance of a new algorithm for estimating sleep and outline the limitations. Based on these results, we conclude that a single device can provide estimates of the sleep interval and total sleep time with sufficient accuracy for the measurement of daily physical activity, sedentary behaviour, and sleep, on a group level in free-living settings.


Influences of evergreen gymnosperm and deciduous angiosperm tree species on the functioning of temperate and boreal forests.
It has been recognized for a long time that the overstorey composition of a forest partly determines its biological and physical-chemical functioning. Here, we review evidence of the influence of evergreen gymnosperm (EG) tree species and deciduous angiosperm (DA) tree species on the water balance, physical-chemical soil properties and biogeochemical cycling of carbon and nutrients. We used scientific publications based on experimental designs where all species grew on the same parent material and initial soil, and were similar in stage of stand development, former land use and current management. We present the current state of the art, define knowledge gaps, and briefly discuss how selection of tree species can be used to mitigate pollution or enhance accumulation of stable organic carbon in the soil. The presence of EGs generally induces a lower rate of precipitation input into the soil than DAs, resulting in drier soil conditions and lower water discharge. Soil temperature is generally not different, or slightly lower, under an EG canopy compared to a DA canopy. Chemical properties, such as soil pH, can also be significantly modified by taxonomic groups of tree species. Biomass production is usually similar or lower in DA stands than in stands of EGs. Aboveground production of dead organic matter appears to be of the same order of magnitude between tree species groups growing on the same site. Some DAs induce more rapid decomposition of litter than EGs because of the chemical properties of their tissues, higher soil moisture and favourable conditions for earthworms. Forest floors consequently tend to be thicker in EG forests compared to DA forests. Many factors, such as litter lignin content, influence litter decomposition and it is difficult to identify specific litter-quality parameters that distinguish litter decomposition rates of EGs from DAs. Although it has been suggested that DAs can result in higher accumulation of soil carbon stocks, evidence from field studies does not show any obvious trend. Further research is required to clarify if accumulation of carbon in soils (i.e. forest floor + mineral soil) is different between the two types of trees. Production of belowground dead organic matter appears to be of similar magnitude in DA and EG forests, and root decomposition rate lower under EGs than DAs. However there are some discrepancies and still are insufficient data about belowground pools and processes that require further research. Relatively larger amounts of nutrients enter the soil-plant biogeochemical cycle under the influence of EGs than DAs, but recycling of nutrients appears to be slightly enhanced by DAs. Understanding the mechanisms underlying forest ecosystem functioning is essential to predicting the consequences of the expected tree species migration under global change. This knowledge can also be used as a mitigation tool regarding carbon sequestration or management of surface waters because the type of tree species affects forest growth, carbon, water and nutrient cycling.


NF-κB c-Rel Is Dispensable for the Development but Is Required for the Cytotoxic Function of NK Cells.
Natural Killer (NK) cells are cytotoxic lymphocytes critical to the innate immune system. We found that germline deficiency of NF-κB c-Rel results in a marked decrease in cytotoxic function of NK cells, both in vitro and in vivo, with no significant differences in the stages of NK cell development. We found that c-Rel binds to the promoters of perforin and granzyme B, two key proteins required for NK cytotoxicity, and controls their expression. We generated a NK cell specific c-Rel conditional knockout to study NK cell intrinsic role of c- Rel and found that both global and conditional c-Rel deficiency leads to decreased perforin and granzyme B expression and thereby cytotoxic function. We also confirmed the role of c-Rel in perforin and granzyme B expression in human NK cells. c-Rel reconstitution rescued perforin and granzyme B expressions in c-Rel deficient NK cells and restored their cytotoxic function. Our results show a previously unknown role of c-Rel in transcriptional regulation of perforin and granzyme B expressions and control of NK cell cytotoxic function.


Reduced fitness and abnormal cardiopulmonary responses to maximal exercise testing in children and young adults with sickle cell anemia.
Physiologic contributors to reduced exercise capacity in individuals with sickle cell anemia (SCA) are not well understood. The objective of this study was to characterize the cardiopulmonary response to maximal cardiopulmonary exercise testing (CPET) and determine factors associated with reduced exercise capacity among children and young adults with SCA. A cross-sectional cohort of 60 children and young adults (mean 15.1 ± 3.4 years) with hemoglobin SS or S/β(0) thalassemia and 30 matched controls (mean 14.6 ± 3.5 years) without SCA or sickle cell trait underwent maximal CPET by a graded, symptom-limited cycle ergometry protocol with breath-by-breath, gas exchange analysis. Compared to controls without SCA, subjects with SCA demonstrated significantly lower peak VO2 (26.9 ± 6.9 vs. 37.0 ± 9.2 mL/kg/min, P < 0.001). Subjects demonstrated slower oxygen uptake (ΔVO2/ΔWR, 9 ± 2 vs. 12 ± 2 mL/min/watt, P < 0.001) and lower oxygen pulse (ΔVO2/ΔHR, 12 ± 4 vs. 20 ± 7 mL/beat, P < 0.001) as well as reduced oxygen uptake efficiency (ΔVE/ΔVO2, 42 ± 8 vs. 32 ± 5, P < 0.001) and ventilation efficiency (ΔVE/ΔVCO2, 30.3 ± 3.7 vs. 27.3 ± 2.5, P < 0.001) during CPET. Peak VO2 remained significantly lower in subjects with SCA after adjusting for age, sex, body mass index (BMI), and hemoglobin, which were independent predictors of peak VO2 for subjects with SCA. In the largest study to date using maximal CPET in SCA, we demonstrate that children and young adults with SCA have reduced exercise capacity attributable to factors independent of anemia. Complex derangements in gas exchange and oxygen uptake during maximal exercise are common in this population.


Stability of CSF metabolites measured by proton NMR.
Analysis of cerebrospinal fluid (CSF) metabolites can provide data regarding CNS involvement in neurologic and psychiatric illness. However, there is lack of research into the effect of processing and storage of CSF specimens on the levels of metabolites analyzed. CSF specimens from 10 depressed patients were analyzed by proton NMR before and after 72 hours exposure to room temperature. No effect of exposure was found on myoinositol, glucose, acetate, and alanine CSF levels and there was a substantial decrease of citrate (>50%) and increase in lactate, glutamine, creatine, and creatinine levels.


Computer-based records. Still searching for the 'Holy Grail'.
The developers of three new databases propose to evaluate computer-based patient record systems in great detail. They promise to rigorously benchmark how CPR systems perform on a suite of tests in what they are calling a 'CPR certification laboratory.'


[Effect of intralesional treatment with emetine hydrochloride on Leishmania (Viannia) braziliensis in hamsters].
The therapeutic effect of the emetine hydrochloride alkaloid administered intralesionally was compared with that of standard parenteral treatment with Glucantime in outbred male hamsters experimentally infected with 4 x 10(3) amastigotes of Leishmania (Viannia) braziliensis. Both chemotherapeutic agents reduced significantly (P < 0.01) the average lesion sizes in experimental animals in comparison with those untreated. The alkaloid infiltration was found to be as effective as the antimonial injection for clinical resolution. The ultrastructural effects on the Leishmania parasites exposed to emetine were observed mainly in the inner cytoplasm, which appeared disorganized, pycnotic and with loss of morphological definition; however, any known emetine hydrochloride action mechanism factor could not be directly related with ultrastructure effects detected on leishmanial parasites. Smears, conventional histopathology, culture in NNN medium and indirect immunoperoxidase method showed viable amastigotes in nodules and/or scars of all the evaluated hamsters 75 to 230 days after the end of treatment. These findings suggest that measurement of the size of cutaneous leishmania lesions does not appear to be a valid criterion for evaluating the efficiency of chemotherapy in experimental LT. Detection of leishmania parasites in the lesion scars, supports the hypothesis that man could be considered as an domestic reservoir.


[Structiure of beta-beta-hairpins closed into cycles by S-S-bridges].
In the present study, a stereochemical analysis of proteins containing beta-beta-hairpins closed into cycles by S-S-bridges has been performed. A database of these proteins has been compiled from the Protein Data Bank (total 428 PDB entries). 390 beta-beta-hairpins closed into cycles by S-S-bridges have been found in non-homologous proteins included into the database. Analysis showed that 118 hairpins contain S-S-bridges formed by cysteins located opposite to each other in the neighboring beta-strands. Among them, 110 hairpins are left-turned and 8 beta-hairpins are right -turned when viewed from the same side where S-S-bridges are located. In the other group of 272 beta-hairpins, the S-S-bridge is formed by two cysteins one of which is located in the beta-strand and the other in the loop juxtaposed to the beta-hairpin at the N- (84%) or C-terminus (16%). As shown, in most cases the loop-hairpin region closed into a cycle by the S-S-bridge formes a turn of a left-handed superhelix.


Targeting DNA Replication Stress for Cancer Therapy.
The human cellular genome is under constant stress from extrinsic and intrinsic factors, which can lead to DNA damage and defective replication. In normal cells, DNA damage response (DDR) mediated by various checkpoints will either activate the DNA repair system or induce cellular apoptosis/senescence, therefore maintaining overall genomic integrity. Cancer cells, however, due to constitutive growth signaling and defective DDR, may exhibit "replication stress" -a phenomenon unique to cancer cells that is described as the perturbation of error-free DNA replication and slow-down of DNA synthesis. Although replication stress has been proven to induce genomic instability and tumorigenesis, recent studies have counterintuitively shown that enhancing replicative stress through further loosening of the remaining checkpoints in cancer cells to induce their catastrophic failure of proliferation may provide an alternative therapeutic approach. In this review, we discuss the rationale to enhance replicative stress in cancer cells, past approaches using traditional radiation and chemotherapy, and emerging approaches targeting the signaling cascades induced by DNA damage. We also summarize current clinical trials exploring these strategies and propose future research directions including the use of combination therapies, and the identification of potential new targets and biomarkers to track and predict treatment responses to targeting DNA replication stress.


Scaling of mutational effects in models for pleiotropy.
Mutation-selection models provide a framework to relate the parameters of microevolution to properties of populations. Like all models, these must be subject to test and refinement in light of experiments. The standard mutation-selection model assumes that the effects of a pleiotropic mutation on different characters are uncorrelated. As a consequence of this assumption, mutations of small overall effect are suppressed. For strong enough pleiotropy, the result is a nonvanishing fraction of a population with the "perfect" phenotype. However, experiments on microorganisms and experiments on protein structure and function contradict the assumptions of the standard model, and Kimura's observations of heterogeneity within populations contradict its conclusions. Guided by these observations, we present an alternative model for pleiotropic mutations. The new model allows mutations of small overall effect and thus eliminates the finite fraction of the population with the perfect phenotype.


Impaired prefrontal sleep spindle regulation of hippocampal-dependent learning in older adults.
A hallmark feature of cognitive aging is a decline in the ability to form new memories. Parallel to these cognitive impairments are marked disruptions in sleep physiology. Despite recent evidence in young adults establishing a role for sleep spindles in restoring hippocampal-dependent memory formation, the possibility that disrupted sleep physiology contributes to age-related decline in hippocampal-dependent learning remains unknown. Here, we demonstrate that reduced prefrontal sleep spindles by over 40% in older adults statistically mediates the effects of old age on next day episodic learning, such that the degree of impaired episodic learning is explained by the extent of impoverished prefrontal sleep spindles. In addition, prefrontal spindles significantly predicted the magnitude of impaired next day hippocampal activation, thereby determining the influence of spindles on post-sleep learning capacity. These data support the hypothesis that disrupted sleep physiology contributes to age-related cognitive decline in later life, the consequence of which has significant treatment intervention potential.


The effects of organizational flexibility on nurse utilization and vacancy statistics in Ontario hospitals.
Social and economic changes in industrial societies during the past quarter-century encouraged organizations to develop greater flexibility in their employment systems in order to adapt to organizational restructuring and labour market shifts (Kallenberg 2003). During the 1990s this trend became evident in healthcare organizations. Before healthcare restructuring, employment in the acute hospital sector was more stable, with higher levels of full-time staff. However, in the downsizing era, employers favoured more flexible, contingent workforces (Zeytinoglu 1999). As healthcare systems evolved, staffing patterns became more chaotic and predicting staffing requirements more complex. Increased use of casual and part-time staff, overtime and agency nurses, as well as alterations in skills mix, masked vacancy counts and thus rendered this measurement of nursing demand increasingly difficult. This study explores flexible nurse staffing practices and demonstrates how data such as nurse vacancy statistics, considered in isolation from nurse utilization information, are inaccurate indicators of nursing demand and nurse shortage. It develops an algorithm that provides a standard methodology for improved monitoring and management of nurse utilization data and better quantification of vacancy statistics. Use of standard methodology promotes more accurate measurement of nurse utilization and shortage. Furthermore, it provides a solid base for improved nursing workforce planning, production and management.


Peroxiredoxin Asp f3 Is Essential for Aspergillus fumigatus To Overcome Iron Limitation during Infection.
Aspergillus fumigatus is an important fungal pathogen that causes allergic reactions but also life-threatening infections. One of the most abundant A. fumigatus proteins is Asp f3. This peroxiredoxin is a major fungal allergen and known for its role as a virulence factor, vaccine candidate, and scavenger of reactive oxygen species. Based on the hypothesis that Asp f3 protects A. fumigatus against killing by immune cells, we investigated the susceptibility of a conditional aspf3 mutant by employing a novel assay. Surprisingly, Asp f3-depleted hyphae were killed as efficiently as the wild type by human granulocytes. However, we identified an unexpected growth defect of mutants that lack Asp f3 under low-iron conditions, which explains the avirulence of the Δaspf3 deletion mutant in a murine infection model. A. fumigatus encodes two Asp f3 homologues which we named Af3l (Asp f3-like) 1 and Af3l2. Inactivation of Af3l1, but not of Af3l2, exacerbated the growth defect of the conditional aspf3 mutant under iron limitation, which ultimately led to death of the double mutant. Inactivation of the iron acquisition repressor SreA partially compensated for loss of Asp f3 and Af3l1. However, Asp f3 was not required for maintaining iron homeostasis or siderophore biosynthesis. Instead, we show that it compensates for a loss of iron-dependent antioxidant enzymes. Iron supplementation restored the virulence of the Δaspf3 deletion mutant in a murine infection model. Our results unveil the crucial importance of Asp f3 to overcome nutritional immunity and reveal a new biological role of peroxiredoxins in adaptation to iron limitation. IMPORTANCE Asp f3 is one of the most abundant proteins in the pathogenic mold Aspergillus fumigatus. It has an enigmatic multifaceted role as a fungal allergen, virulence factor, reactive oxygen species (ROS) scavenger, and vaccine candidate. Our study provides new insights into the cellular role of this conserved peroxiredoxin. We show that the avirulence of a Δaspf3 mutant in a murine infection model is linked to a low-iron growth defect of this mutant, which we describe for the first time. Our analyses indicated that Asp f3 is not required for maintaining iron homeostasis. Instead, we found that Asp f3 compensates for a loss of iron-dependent antioxidant enzymes. Furthermore, we identified an Asp f3-like protein which is partially functionally redundant with Asp f3. We highlight an unexpected key role of Asp f3 and its partially redundant homologue Af3l1 in overcoming the host's nutritional immunity. In addition, we uncovered a new biological role of peroxiredoxins.


Protein synthesis in the liver of rats injected with cholesteryl 14-methylhexadecanoate.
1. Rats were injected intraperitoneally with cholesteryl 14-methylhexadecanoate and killed after various intervals of time up to 3 days; ribosomes and cell sap were isolated from their liver tissue. These fractions were tested for their ability to participate in protein synthesis. 2. Protein synthesis in complete systems containing ribosomes, cell sap and all necessary cofactors was significantly enhanced at 12 and 72h after the injection and significantly inhibited at 24h. At early times after injection isolated ribosomes had a slightly enhanced ability to bind nRNA. Peptide-elongation processes (i.e. binding of aminoacyl-tRNA to ribosomes, peptidyl transfer and polyphenylalanine synthesis) showed significant stimulation or inhibition depending on the time after injection of the ester. 3. A correlation was found between the ability of cell sap to stimulate polyphenylalanine synthesis and the relative cholesteryl 14-methylhexadecanoate content in the postmicrosomal supernatant at different time-intervals after administration of the ester. No significant changes were found in its content in the whole liver tissue. 4. Since the injected ester has previously been shown to accumulate in some enzymic fractions, the changes in its relative content may represent a regulatory mechanism modulating the rate of protein synthesis.


Localization of viral protein in avian-encephalomyelitis-virus-infected hens.
Eighteen 2-year-old hens were infected orally with wild avian encephalomyelitis virus and examined using the immunofluorescent-antibody technique for 30 days. Viral antigen was detected in 10 organ tissues (liver, spleen, kidney, pancreas, and ovary) in 7 birds during the first half of the observation period and sporadically in alimentary tracts of 12 birds throughout this period, especially the lower parts of the intestines, but it was not detected in other organ tissues. Thirteen test-positive hens showed more localization of the antigen than baby chicks of other studies did.


Targeted Thrombospondin-1 Expression in Ocular Vascular Development and Neovascularization.
Tight regulation of positive and negative regulators of angiogenesis is essential, particularly in the eye where their dysregulation can lead to vision loss. Thrombospondin-1 (TSP1) is a matricellular protein that negatively regulates angiogenesis and inflammation in the eye. It aids ocular vascular homeostasis such that its loss contributes to increased retinal vascular density and pathologic ocular neovascularization. Our previous studies demonstrated that mice globally lacking TSP1 expression had increased retinal vascular density, decreased hyperoxia-induced retinal vessel loss, and increased choroidal neovascularization. Here we determined the impact to the ocular vasculature of endothelial cell, pericyte, or astrocyte loss of TSP1 expression. Only lack of TSP1 expression in endothelial cells was sufficient to increase choroidal neovascularization with mice lacking expression in pericytes or astrocytes not demonstrating a significant impact. Although the global TSP1 knockout mice demonstrated increased retinal vascular density, individual cell type loss of TSP1 resulted in decreased retinal endothelial cell numbers before and/or after vascular maturation in a cell type specific fashion. Retinas from mice lacking TSP1 expression in endothelial cells, pericytes or astrocytes were not protected from retinal vessel regression in response to hyperoxia as we previously observed in the global knockout. Thus, modulation of TSP1 expression in individual cell types demonstrates a response that is unique to the role TSP1 plays in that cell type of interest, and their coordinated activity is critical for vision.


[Electrophoretic forms of glucose-6-phosphate dehydrogenase, acid phosphatase and esterase in Amoeba species amoebas].
Glucose-6-phosphate dehydrogenase (G6PD), acid phosphatase and esterases in free-living amoebae of 7 Amoeba species were investigated with the use of disc-electrophoresis in polyacrylamide gel. The evidence provided is suggestive that the electrophoretic isoenzyme patterns of acid phosphatase and esterases (and G6PD in some cases), in addition to a few morphological characters, can serve as a taxonomic criterion for species identification within this genus, as well as for revealing erroneously classified species and strains. It is suggested that A. indica is an independent species whose preliminary diagnosis has been given in this paper. It is concluded that A. discoides and A. lescherae are strains of A. proteus, rather than two independent species. A and As-102 amoebian strains, kept in the collection of protozoan strains and species of the Institute of Cytology RAS and referred to as strains of A. proteus, belong in reality to another Amoeba species and even to another genus within the family Amoebidae. This conclusion has been documented by results of our analysis of electrophoretic patterns of acid phosphatase and esterases in these strains.


The multi-year cumulative effects of alternative stocking rate and grazing management practices on pasture productivity and utilization efficiency.
The production and utilization of increased quantities of high quality pasture is of paramount importance in pasture-based milk production systems. The objective of this study was to evaluate the cumulative effects of alternative integrated grazing strategies, incorporating alternative stocking rate (SR) and grazing severities, on pasture productivity and grazing efficiency over multiple years within farm systems using perennial ryegrass dominant pastures. Three whole-farm SR treatments were compared over 4 complete grazing seasons (2009 to 2012 inclusive): low (2.51 cows/ha; LSR), medium (2.92 cows/ha; MSR), and high (3.28 cows/ha; HSR). Each system had its own farmlet containing 18 paddocks and remained on the same treatment for the duration of the study. Stocking rate had a significant effect on all grazing variables with the exception of soil fertility status and sward density. Increased SR resulted in increased total annual net pasture accumulation, improved sward nutritive value, and increased grazed pasture utilization. Total annual net pasture accumulation was greatest in HSR [15,410kg of dry matter (DM)/ha], intermediate for MSR (14,992kg of DM/ha), and least for LSR (14,479kg of DM/ha) during the 4-yr study period. A linear effect of SR on net pasture accumulation was detected with an increase in net pasture accumulation of 1,164.4 (SE=432.7) kg of DM/ha for each 1 cow/ha increase in SR. Pregrazing pasture mass and height and postgrazing residual pasture mass and height were greatest for LSR, intermediate for the MSR, and lowest for the HSR. In comparison with the LSR, the imposition of a consistently increased grazing severity coupled with increased whole farm SR in MSR and HSR treatments arrested the decline in sward nutritive value, typically observed during mid-season. Incorporating the individual beneficial effects of SR on pasture accumulation, nutritive value, and utilization efficiency, total proportional energy (unité fourragère lait) utilization per hectare increased significantly with increasing SR (+0.026 and +0.081 for MSR and HSR, respectively). These results quantify the significant effect of grazing management practices on the feed production capability of modern perennial ryegrass pastures for intensive grazing dairy production systems. Furthermore, these results highlight the importance of consistently imposing grazing treatments over multiple years, and within integrated whole farm systems, to accurately assess the longer term effects of alternate grazing management practices on pasture productivity.


Amino Acid and Sugar Catabolism in the Marine Bacterium Phaeobacter inhibens DSM 17395 from an Energetic Viewpoint.
Growth energetics and metabolic efficiency contribute to the lifestyle and habitat imprint of microorganisms. Roseobacters constitute one of the most abundant and successful marine bacterioplankton groups. Here, we reflect on the energetics and metabolic efficiency of Phaeobacter inhibens DSM 17395, a versatile heterotrophic roseobacter. Fourteen different substrates (five sugars and nine amino acids) and their degradation pathways were assessed for energetic efficiencies based on catabolic ATP yields, calculated from net formed ATP and reducing equivalents. The latter were converted into ATP by employing the most divergent coupling ratios (i.e., ions per ATP) currently known for F1Fo ATP synthases in heterotrophic bacteria. The catabolic ATP yields of the pathways studied in P. inhibens differed ∼3-fold. The actual free energy costs for ATP synthesis were estimated at 81.6 kJ per mol ATP (3.3 ions per ATP) or 104.2 kJ per mol ATP (4.3 ions per ATP), yielding an average thermodynamic efficiency of ∼37.7% or ∼29.5%, respectively. Growth performance (rates, yields) and carbon assimilation efficiency were determined for P. inhibens growing in process-controlled bioreactors with 10 different single substrates (Glc, Man, N-acetylglucosamine [Nag], Phe, Trp, His, Lys, Thr, Val, or Leu) and with 2 defined substrate mixtures. The efficiencies of carbon assimilation into biomass ranged from ∼28% to 61%, with His/Trp and Thr/Leu yielding the lowest and highest levels. These efficiencies correlated with catabolic and ATP yields only to some extent. Substrate-specific metabolic demands and/or functions, as well as the compositions of the substrate mixtures, apparently affected the energetic costs of growth. These include energetic burdens associated with, e.g., slow growth, stress, and/or the production of tropodithietic acid.IMPORTANCE Heterotrophic members of the bacterioplankton serve the marine ecosystem by transforming organic matter, an activity that is governed by the bacterial growth efficiencies (BGEs) obtained under given environmental conditions. In marine ecology, the concept of BGE refers to the carbon assimilation efficiency within natural communities. The marine bacterium studied here, Phaeobacter inhibens DSM 17395, is a copiotrophic representative of the globally abundant Roseobacter group, and the 15 catabolic pathways investigated are widespread among these marine heterotrophs. Combining pathway-specific catabolic ATP yields with in-depth quantitative physiological data could (i) provide a new baseline for the study of growth energetics and efficiency in further Roseobacter group members and other copiotrophic marine bacteria in productive coastal ecosystems and (ii) contribute to a better understanding of the factors controlling BGE (including the additional energetic burden arising from widespread secondary-metabolite formation) based on laboratory studies with pure cultures.