Quantitative anatomy of the large variant right hepatic vein: A systematic three-dimensional analysis.
Anatomical variations of the right hepatic vein, especially large variant right hepatic veins (≥5 mm), have important clinical implications in liver transplantation and resection. This study aimed to evaluate anatomical variations of the right hepatic vein using quantitative three-dimensional visualization analysis. Computed tomography images of 650 patients were retrospectively analyzed, and three-dimensional visualization was applied using the derived data to analyze large variant right hepatic veins. The proportion of the large variant right hepatic vein was 16.92% (110/650). According to the location and number of the variant right hepatic veins, the configuration of the right hepatic venous system was divided into seven subtypes. The length of the retrohepatic inferior vena cava had a positive correlation with the diameter of the right hepatic vein (rs = 0.266, p = 0.001) and the variant right hepatic veins (rs = 0.211, p = 0.027). The diameter of the right hepatic vein was positively correlated with that of the middle hepatic vein (rs = 0.361, p < 0.001), while it was inversely correlated with that of the variant right hepatic veins (rs = -0.267, p = 0.005). The right hepatic vein diameter was positively correlated with the drainage volume (rs = 0.489, p < 0.001), while the correlation with the variant right hepatic veins drainage volume was negative (rs = -0.460, p < 0.001). The number of the variant right hepatic veins and their relative diameters were positively correlated (p < 0.001). The volume and percentage of the drainage area of the right hepatic vein decreased significantly as the number of the variant right hepatic vein increased (p < 0.001). The findings of this study concerning the variations of the hepatic venous system may be useful for the surgical planning of liver resection or transplantation.


Chaperonins from an Antarctic archaeon are predominantly monomeric: crystal structure of an open state monomer.
Archaea are abundant in permanently cold environments. The Antarctic methanogen, Methanococcoides burtonii, has proven an excellent model for studying molecular mechanisms of cold adaptation. Methanococcoides burtonii contains three group II chaperonins that diverged prior to its closest orthologues from mesophilic Methanosarcina spp. The relative abundance of the three chaperonins shows little dependence on organism growth temperature, except at the highest temperatures, where the most thermally stable chaperonin increases in abundance. In vitro and in vivo, the M. burtonii chaperonins are predominantly monomeric, with only 23-33% oligomeric, thereby differing from other archaea where an oligomeric ring form is dominant. The crystal structure of an N-terminally truncated chaperonin reveals a monomeric protein with a fully open nucleotide binding site. When compared with closed state group II chaperonin structures, a large-scale ≈ 30° rotation between the equatorial and intermediate domains is observed resulting in an open nucleotide binding site. This is analogous to the transition observed between open and closed states of group I chaperonins but contrasts with recent archaeal group II chaperonin open state ring structures. The predominance of monomeric form and the ability to adopt a fully open nucleotide site appear to be unique features of the M. burtonii group II chaperonins.


Hair loss due to lichen planopilaris after hair transplantation: a report of two cases and a literature review.
Androgenetic alopecia is often treated by follicular unit transplantation, a technique that involves minimal risk of hair loss because of the more resistant nature of the donor area. Lichen planopilaris is a cicatricial alopecia that causes permanent destruction of hair follicles. We report two cases of post-transplantation lesions compatible with lichen planopilaris in both recipient and donor areas. The quality of the hair follicles in the donor area was apparently compromised by lichen planopilaris, the probable cause of hair loss. Similar reports are rare. When lichen planopilaris is suspected, a biopsy of the scalp must be performed to avoid transplantation during disease activity.


Repeat bedside percutaneous dilational tracheostomy is a safe procedure.
Previous tracheostomy has been considered a relative contraindication for percutaneous dilational tracheostomy. The objective of this study was to assess the safety of percutaneous dilational tracheostomy in critically ill patients with a history of previous tracheostomy. Retrospective, single-center case series of all consecutive patients requiring repeat tracheostomy for continued mechanical ventilatory support. Intensive care unit of a tertiary-care referral center. Fourteen patients (eight female, six male) with a median age of 70 yrs (range, 33-94). All patients had previously undergone tracheostomy. Bedside percutaneous dilational tracheostomy. Subjects' previous tracheostomies dated back between 10 days and 8 yrs. Present intubation time before percutaneous dilational tracheostomy varied between 4 and 30 days. Bedside percutaneous dilational tracheostomy was performed successfully in all 14 patients by trained pulmonologists and surgeons. Eleven patients received an 8-mm and three received a 7-mm tracheostomy tube. There were no significant periprocedural complications, and no patient required surgical revision. The only postprocedural complication was accidental decannulation in one patient, which was managed with repeat percutaneous dilational tracheostomy. Trained physicians can safely perform bedside percutaneous dilational tracheostomy after previous tracheostomy. Percutaneous dilational tracheostomy offers an alternative to surgical tracheostomy in this particular patient population and should not be considered contraindicated.


Medial malleolar stress fracture in athletes: diagnosis and operative treatment.
Stress fractures are common overuse injuries in athletes. Medial malleolar stress fractures are rare but they have an important clinical relevance because of their tendency to cause considerable disability and loss of time in sports without proper treatment. The diagnosis of medial malleolar stress fracture is often a challenge and it is therefore often delayed which may cause even further problems in the treatment. The purpose of this study was to increase the awareness of medial malleolar stress fractures as a possible cause for medial ankle pain and to stress the importance of MRI in the diagnostics as well as to evaluate the results of surgical treatment. Between 1995 and 2008, a total of ten athletes with a medial malleolar stress fracture were operated at our centre. All operated cases during those years were included in the study. The cases were retrospectively analyzed. Return to pre-injury level of sport was evaluated and considered as an indicator of successful treatment. Initially all standard radiographs were negative whereas in MRI the fractures were all visible. After operative treatment all medial malleolar stress fractures healed clinically in three to four months and all except one of the athletes were able to return to their pre-injury level of sports. Early MRI is recommended if a medial malleolar stress fracture is suspected. Surgical treatment seems to result in rapid healing of the fracture and return to sports. In our opinion early surgery should be considered especially in athletes.


Relationship of HeLa cell infectivity to biochemical, serological, and virulence characteristics of Yersinia enterocolitica.
Examination of 69 strains of Yersinia enterocolitica which represented 20 serotypes and nontypable isolates for HeLa cell infectivity by a roller tube technique that provided a quantitative index of infection showed that infectivity (index, greater than 3.50) was confined to strains of serotypes O:8, O:3, O:5, 27, O:9, O:1, O:1,2,3, O:2,3, O:4,32, and O:21. All strains that were HeLa positive were sucrose positive and negative for salicin, esculin, rhamnose, raffinose, melibiose, alpha-methylglucoside, and citrate. All HeLa-negative strains were either sucrose and salicin positive or were sucrose negative. Twenty-one strains were examined for virulence by the ability to produce guinea pig conjunctivitis and diarrhea in mice. Positive strains were limited to those that were HeLa positive and were autoagglutination positive and calcium dependent at 35 degrees C. There was no association between virulence and the ability to produce enterotoxin measured by the infant mouse assay. Loss of autoagglutinability and calcium dependency was accompanied by loss of virulence, but HeLa cell infectivity was unchanged. The results suggest that at least two properties are necessary for virulence: the presence of the V and W antigens, mediated by the same plasmid as autoagglutination and calcium dependency, and an invasive factor demonstrated in vitro by HeLa cell infectivity. These virulence properties are found in only certain biotypes of Y. enterocolitica.


Experimental dengue virus challenge of human subjects previously vaccinated with live attenuated tetravalent dengue vaccines.
Protection against dengue requires immunity against all 4 serotypes of dengue virus (DENV). Experimental challenge may be useful in evaluating vaccine-induced immunity. Ten subjects previously vaccinated with a live attenuated tetravalent dengue vaccine (TDV) and 4 DENV-naive control subjects were challenged by subcutaneous inoculation of either 10(3) plaque-forming units (PFU) of DENV-1 or 10(5) PFU of DENV-3. Two additional subjects who did not develop DENV-3 neutralizing antibody (NAb) from TDV were revaccinated with 10(4) PFU of live attenuated DENV-3 vaccine to evaluate memory response. All 5 TDV recipients were protected against DENV-1 challenge. Of the 5 TDV recipients challenged with DENV-3, 2 were protected. All DENV-3-challenge subjects who developed viremia also developed elevated liver enzyme levels, and 2 had values that were >10 times greater than normal. Of the 2 subjects revaccinated with DENV-3 vaccine, 1 showed a secondary response to DENV-2, while neither showed such response to DENV-3. All 4 control subjects developed dengue fever from challenge. Protection was associated with presence of NAb, although 1 subject was protected despite a lack of measurable NAb at the time of DENV-1 challenge. Vaccination with TDV induced variable protection against subcutaneous challenge. DENV-3 experimental challenge was associated with transient but marked elevations of transaminases.


Disposable ultrasound-sensing chronic cranial window by soft nanoimprinting lithography.
Chronic cranial window (CCW) is an essential tool in enabling longitudinal imaging and manipulation of various brain activities in live animals. However, an active CCW capable of sensing the concealed in vivo environment while simultaneously providing longitudinal optical access to the brain is not currently available. Here we report a disposable ultrasound-sensing CCW (usCCW) featuring an integrated transparent nanophotonic ultrasonic detector fabricated using soft nanoimprint lithography process. We optimize the sensor design and the associated fabrication process to significantly improve detection sensitivity and reliability, which are critical for the intend longitudinal in vivo investigations. Surgically implanting the usCCW on the skull creates a self-contained environment, maintaining optical access while eliminating the need for external ultrasound coupling medium for photoacoustic imaging. Using this usCCW, we demonstrate photoacoustic microscopy of cortical vascular network in live mice over 28 days. This work establishes the foundation for integrating photoacoustic imaging with modern brain research.


In vitro assessments of nanoplexes of polyethylenimine-coated graphene oxide-plasmid through various cancer cell lines and primary mesenchymal stem cells.
Efficient gene therapy relies on an efficient gene delivery system. Viral gene delivery approaches excel in transferring and expressing external genes; however, their immunogenicity and difficulty in large-scale production limit their clinical applications. In contrast, nanoparticle-based gene delivery systems have gained increasing attention due to less immunogenicity and more convenience for large-scale production. Nevertheless, their poor transfection efficiency compared to viral systems remains a significant obstacle. In the present study, we investigated the transfection efficiency of our PEI-coated graphene oxides in HEK293T, Calu-3, Calu-6 cell lines, and primary human bone marrow mesenchymal stem cell (MSC). The high surface ratio and good biocompatibility of graphene oxide make it an appealing tool for gene delivery systems. However, the low dispersity of graphene oxide in aqueous environments is the first barrier that needs to be conquered. For this, we enhanced the dispersity and stability of graphene oxide in water by sonicating it for at least 5 hours at a pH of 7. Then, graphene oxide was conjugated with branched PEI (25 kDa) to have a positive charge, enabling it to condense nucleic acids with a naturally negative potential. The physio-chemical characteristics of our synthesized nano-carriers (GO-PEI) were determined by DLS, FT-IR, and AFM. The utilized plasmid in polyplexes contained a GFP gene, allowing us to verify transfection efficiency through fluorescent microscopy and flow cytometry. While GO-PEI carriers were highly efficient in transfecting HEK293T cells, the transfection efficiency in MSCs and Calu-3 cells was notably low. We suppose that the main reason for the low transfection efficiency of GO-PEI in these cells is due to its higher toxicity. Despite this, considering the various advantages of graphene oxide in drug delivery as well as its optical and electrical applications in biomedicine, we propose to functionalize graphene oxide with more biocompatible materials to enhance its potential as a successful gene carrier in these cell types.


PD-L1/B7H-1 inhibits the effector phase of tumor rejection by T cell receptor (TCR) transgenic CD8+ T cells.
Although increased circulating tumor antigen-specific CD8(+) T cells can be achieved by vaccination or adoptive transfer, tumor progression nonetheless often occurs through resistance to effector function. To develop a model for identifying mechanisms of resistance to antigen-specific CTLs, poorly immunogenic B16-F10 melanoma was transduced to express the K(b)-binding peptide SIYRYYGL as a green fluorescent protein fusion protein that should be recognized by high-affinity 2C TCR transgenic T cells. Although B16.SIY cells expressed high levels of antigen and were induced to express K(b) in response to IFN-gamma, they were poorly recognized by primed 2C/RAG2(-/-) T cells. A screen for candidate inhibitory ligands revealed elevated PD-L1/B7H-1 on IFN-gamma-treated B16-F10 cells and also on eight additional mouse tumors and seven human melanoma cell lines. Primed 2C/RAG2(-/-)/PD-1(-/-) T cells showed augmented cytokine production, proliferation, and cytolytic activity against tumor cells compared with wild-type 2C cells. This effect was reproduced with anti-PD-L1 antibody present during the effector phase but not during the priming culture. Adoptive transfer of 2C/RAG2(-/-)/PD-1(-/-) T cells in vivo caused tumor rejection under conditions in which wild-type 2C cells or CTLA-4-deficient 2C cells did not reject. Our results support interfering with PD-L1/PD-1 interactions to augment the effector function of tumor antigen-specific CD8(+) T cells in the tumor microenvironment.


The depth and breadth of multiple perceptual asymmetries in right handers and non-right handers.
Several non-verbal perceptual and attentional processes have been linked with specialization of the right cerebral hemisphere. Given that most people have a left hemispheric specialization for language, it is tempting to assume that functions of these two classes of dominance are related. Unfortunately, such models of complementarity are notoriously hard to test. Here we suggest a method which compares frequency of a particular perceptual asymmetry with known frequencies of left hemispheric language dominance in right-handed and non-right handed groups. We illustrate this idea using the greyscales and colourscales tasks, chimeric faces, emotional dichotic listening, and a consonant-vowel dichotic listening task. Results show a substantial "breadth" of leftward bias on the right hemispheric tasks and rightward bias on verbal dichotic listening. Right handers and non-right handers did not differ in terms of proportions of people who were left biased for greyscales/colourscales. Support for reduced typical biases in non-right handers was found for chimeric faces and for CV dichotic listening. Results are discussed in terms of complementary theories of cerebral asymmetries, and how this type of method could be used to create a taxonomy of lateralized functions, each categorized as related to speech and language dominance, or not.


DNA Methylation and Protein Markers of Chronic Inflammation and Their Associations With Brain and Cognitive Aging.
To investigate chronic inflammation in relation to cognitive aging by comparison of an epigenetic and serum biomarker of C-reactive protein and their associations with neuroimaging and cognitive outcomes. At baseline, participants (n = 521) were cognitively normal, around 73 years of age (mean 72.4, SD 0.716), and had inflammation, vascular risk (cardiovascular disease history, hypertension, diabetes, smoking, alcohol consumption, body mass index), and neuroimaging (structural and diffusion MRI) data available. Baseline inflammatory status was quantified by a traditional measure of peripheral inflammation-serum C-reactive protein (CRP)-and an epigenetic measure (DNA methylation [DNAm] signature of CRP). Linear models were used to examine the inflammation-brain health associations; mediation analyses were performed to interrogate the relationship between chronic inflammation, brain structure, and cognitive functioning. We demonstrate that DNAm CRP shows significantly (on average 6.4-fold) stronger associations with brain health outcomes than serum CRP. DNAm CRP is associated with total brain volume (β = -0.197, 95% confidence interval [CI] -0.28 to -0.12, p FDR = 8.42 × 10-6), gray matter volume (β = -0.200, 95% CI -0.28 to -0.12, p FDR = 1.66 × 10-5), and white matter volume (β = -0.150, 95% CI -0.23 to -0.07, p FDR = 0.001) and regional brain atrophy. We also find that DNAm CRP has an inverse association with global and domain-specific (speed, visuospatial, and memory) cognitive functioning and that brain structure partially mediates this CRP-cognitive association (up to 29.7%), dependent on lifestyle and health factors. These results support the hypothesis that chronic inflammation may contribute to neurodegenerative brain changes that underlie differences in cognitive ability in later life and highlight the potential of DNAm proxies for indexing chronic inflammatory status. This study provides Class II evidence that a DNAm signature of CRP levels is more strongly associated with brain health outcomes than serum CRP levels.


Use of Intracardiac Compared With Transesophageal Echocardiography for Left Atrial Appendage Occlusion in the Amulet Observational Study.
The aim of this study was to compare patient and procedural characteristics, clinical events, and left atrial appendage (LAA) occlusion rates among implantation procedures guided either by intracardiac echocardiography (ICE) or by transesophageal echocardiography (TEE) in the Amulet Observational Study. TEE is standard for intraprocedural imaging during LAA occlusion. ICE is an alternative enabling local rather than general anesthesia. The Amulet Observational Study is a prospective, multicenter, post-market study of the Amulet device in patients with atrial fibrillation at risk for thromboembolism from the LAA. Procedures were guided by ICE in the left atrium or TEE. A clinical events committee adjudicated serious adverse events. LAA closure was assessed by a core laboratory 1 to 3 months post-implantation. Nine hundred fifty-five procedures were TEE guided and 130 (12%) were ICE guided. Local anesthesia was used in 93% of ICE-guided cases. Implantation success was >99% in both groups (p = 1.00). The procedure was longer (40 ± 31 min vs. 33 ± 21 min; p = 0.01) and contrast use higher (145 ± 157 ml vs. 98 ± 76 ml; p < 0.01) for ICE cases. LAA closure rate was 100% using ICE and 98% using TEE. Vascular access serious adverse events occurred in 0.8% of ICE patients and 1.5% of TEE patients (p = 0.52). Pericardial effusions or tamponade was observed at rates of 2.5% per year and 1.7% per year in ICE and TEE patients, respectively (p = 0.57). Rates of ischemic stroke (4.1% per year vs. 2.6% per year; p = 0.37) and major bleeding (8.2% per year vs. 10.6% per year; p = 0.44) did not differ significantly between ICE and TEE patients. Amulet LAA occlusion procedures guided by ICE and TEE resulted in comparable clinical event and LAA closure rates, without differences in procedural or vascular complications despite longer procedures and more contrast use.


One for the road! A study to assess the efficacy of single low-dose regimen of rasburicase in controlling hyperuricaemia in patients with tumour lysis syndrome due to haematological malignancies.
We conducted a retrospective audit of six patients with various haematological malignancies (two acute lymphoblastic leukaemia, one acute myeloid leukaemia, and three non-Hodgkin lymphoma); these patients were eligible to receive rasburicase, being at high risk of development of tumour lysis syndrome (TLS). They received a fixed single low-dose regimen of rasburicase (7.5 mg) mainly due to financial restriction, as patients were not supported by the National Health Service and did not have health insurance. We compared uric acid, creatinine levels, and electrolytes (i.e. phosphate, potassium, and calcium) before and after rasburicase administration and also assessed the need for renal replacement therapy after treatment. All six patients had a significant reduction in uric acid levels on the first day, achieving a response rate of 100% (p = 0.008994); creatinine, phosphate, and potassium were reduced significantly as well, with the p values of 0.0439, 0.014326, and 0.002008, respectively; only one patient needed renal replacement therapy in the form of haemodialysis, due to concerns about hyperphosphataemia. Financial difficulties faced either because patients lacked insurance or because of the restricted National Health Service budget in Egypt have resulted in the unavailability of certain modalities of treatment in cancer care and the need to consider more economic yet efficient approaches. Our experience suggests that a single low-dose rasburicase injection (7.5 mg) is an efficient and cost-effective method to control hyperuricaemia in patients with a high risk of developing TLS when compared with the more expensive and extended standard regimen and doses recommended.


Who's really hypertensive?--Quality control issues in the assessment of blood pressure for randomized trials.
The characterization of blood pressure in treatment trials assessing the benefits of blood pressure lowering regimens is a critical factor for the appropriate interpretation of study results. With numerous operators involved in the measurement of blood pressure in many thousands of patients being screened for entry into clinical trials, it is essential that operators follow pre-defined measurement protocols involving multiple measurements and standardized techniques. Blood pressure measurement protocols have been developed by international societies and emphasize the importance of appropriate choice of cuff size, identification of Korotkoff sounds, and digit preference. Training of operators and auditing of blood pressure measurement may assist in reducing the operator-related errors in measurement. This paper describes the quality control activities adopted for the screening stage of the 2nd Australian National Blood Pressure Study (ANBP2). ANBP2 is cardiovascular outcome trial of the treatment of hypertension in the elderly that was conducted entirely in general practices in Australia. A total of 54 288 subjects were screened; 3688 previously untreated subjects were identified as having blood pressure >140/90 mmHg at the initial screening visit, 898 (24%) were not eligible for study entry after two further visits due to the elevated reading not being sustained. For both systolic and diastolic blood pressure recording, observed digit preference fell within 7 percentage points of the expected frequency. Protocol adherence, in terms of the required minimum blood pressure difference between the last two successive recordings, was 99.8%. These data suggest that adherence to blood pressure recording protocols and elimination of digit preferences can be achieved through appropriate training programs and quality control activities in large multi-centre community-based trials in general practice. Repeated blood pressure measurement prior to initial diagnosis and study entry is essential to appropriately characterize hypertension in these elderly patients.


[Somatization of erectile disorders by the introduction of sildenafil].
With the introduction of the erection-supporting oral drug sildenafil (Viagra) the development of somatic treatments of male sexual dysfunction has reached a temporary peak. The availability of a medicament that is easy to take will result in an increase of the incidence and possibly also the prevalence of erectile disorder. It may even lead to the myth that only a perfect erection is normal. The question is where to draw the line between a normal inconvenience of life and a serious disease. The erection pill will probably be preferred to all other treatments. Therefore, most patients will consult their family physician, who will refer only difficult cases to urologists or sexologists. Physicians should take 'the man behind the penis' into consideration before writing a prescription, otherwise a distressed man without an erection might become a distressed man with an erection. In the long run, psychosexual therapy is less expensive than taking sildenafil for many years.


Properties of short-term synaptic depression at larval neuromuscular synapses in wild-type and temperature-sensitive paralytic mutants of Drosophila.
The larval neuromuscular synapse of Drosophila serves as an important model for genetic and molecular analysis of synaptic development and function. Further functional characterization of this synapse, as well as adult neuromuscular synapses, will greatly enhance the impact of this model system on our understanding of synaptic transmission. Here we describe a form of short-term synaptic depression observed at larval, but not adult, neuromuscular synapses and explore the underlying mechanisms. Larval neuromuscular synapses exhibited a form of short-term depression that was strongly dependent on stimulation frequency over a narrow range of low frequencies (0.1-1 Hz). This form of synaptic depression, referred to here as low-frequency short-term depression (LF-STD), results from an activity-dependent reduction in neurotransmitter release. However, in contrast to the predictions of depletion models, the degree of depression was independent of the initial level of neurotransmitter release over a range of extracellular calcium concentrations. This conclusion was confirmed in two temperature-sensitive (TS) paralytic mutants, cacophony and shibire, which exhibit reduced neurotransmitter release resulting from conditional disruption of presynaptic calcium channels and dynamin, respectively. Higher stimulation frequencies (40 or 60 Hz) produced two components of depression that appeared to include LF-STD as well as a more conventional component of short-term depression. These findings reveal novel properties of short-term synaptic depression and suggest that complementary genetic analysis of larval and adult neuromuscular synapses will further define the in vivo mechanisms of neurotransmitter release and short-term synaptic plasticity.


Reduction and s-carboxymethylation of proteins: microscale method.
INTRODUCTIONReduction and alkylation of microscale amounts of protein (10-50 μg) require special attention. Classical methods for desalting and dialysis should be avoided due to large sample losses. Adding a small quantity of detergent to the protein solution during all manipulations can also reduce protein loss. This protocol describes a procedure for reduction and S-carboxymethylation of low-microgram amounts of protein. Samples are reduced and alkylated in small volumes (<200 μl) in the presence of detergent and desalted (and concentrated) by RP-HPLC in yields of >90%.


Tubulointerstitial responses in the progression of glomerular diseases: albuminuria modulates alpha v beta 5 integrin.
Proteinuria represents one of the most unfavorable prognostic factors in the progression of nephropathies. Several lines of evidence support a role for proteinuria per se in the development of interstitial fibrosis, albeit the molecular mechanisms are still unknown. We investigated the potential role of integrins expressed on tubular cells in regulating the synthesis and organization of interstitial matrix or as mediators of tubulointerstitial damage in conditions mimicking the nephrotic milieu. Under basal conditions, cultured tubular cells highly expressed alpha 3 beta 1 and, at focal contacts, alpha v beta 3. In contrast, alpha v beta 5 was weakly and diffusely distributed all over the plasma membrane. Cultures on a variety of matrix substrates (fibronectin, laminin, collagen types I and IV, vitronectin, von Willebrand factor, fibrinogen) did not induce any phenotypic change in integrin expression by tubular cells. Conversely, the addition of albumin resulted in a highly increased membrane expression of beta 5, which was organized in typical focal contacts and was related to the dose of albumin added. Immunofluorescence, flow cytometry, immunoprecipitation and RT-PCR experiments argue for a complex mechanism that includes increased post-transcriptionally regulated protein synthesis, accelerated conversion of precursors to mature forms, and increased surface delivery to discrete adhesive structures. Up-regulation of the beta 5 chain in tubular cells was confirmed in 9 out of 11 kidney biopsies from proteinuric glomerulonephritides including membranous and focal sclerosing glomerulonephritis, while it was not expressed in nonproteinuric kidneys including five biopsy specimens. This is the first report indicating that proteinuria up-regulates the surface expression and distribution of a specific integrin chain on tubular cells. These observations suggest the participation of integrins in a hitherto unexplored mechanism of tubulo-interstitial responses to glomerular injury.


Circulating immune complexes in the sera and ascites of hepatocellular carcinoma or chronic hepatitis patients.
Circulating immune complexes (CIC) in the sera or ascites of hepatocellular carcinoma (HCC), chronic hepatitis patients and normal healthy persons were measured by polyethylene glycol (PEG) and C1q solid-phase microassay (C1q-SPMA). Both the PEG and C1q-SPMA methods showed the serum CIC levels of HCC patients were significantly higher than those of chronic hepatitis patients and of normal persons. The CIC levels of chronic hepatitis patients were also significantly higher than those of normal persons as detected by PEG method but not by C1q-SPMA. The ascites from HCC patients also had CIC. But the amount of CIC in ascites was significantly lower than those of the serum from the same HCC patients. These results suggest that the increase of CIC may play some pathological role in the HCC patients.


The Effect of Telehealth Interventions on Quality of Life of Cancer Patients: A Systematic Review and Meta-Analysis.
In 2016, ∼1.7 million new cases of cancer were diagnosed. Cancer patients can have physical, functional, and psychosocial issues when dealing with cancer treatment. Telehealth has been effectively introduced to help deliver treatment to patients suffering from chronic disease; however, there is little consensus on its effectiveness in administering sociobehavioral cancer treatments. Thus, this study determines the benefits of telehealth-based interventions providing emotional and symptom support in improving quality of life (QOL) among cancer patients. Two researchers conducted comprehensive searches on PubMed, SCOPUS, Medline, PsycINFO, ERIC, Psychology and Behavioral Collection, and Medline Complete. Key search terms included telehealth or telemedicine and QOL and cancer. Articles were included if they assessed a telehealth-delivered intervention for adult cancer patients and provided a QOL assessment. Data were extracted to calculate mean effect sizes for QOL measures on the effectiveness of telehealth relative to usual care (UC) for cancer treatments. Out of 414 articles identified in our initial search, nine articles fit our inclusion criteria. Both telehealth (Hedges g = 0.211, p = 0.016) and standard of care (Hedges g = 0.217, p < 0.001) cancer treatment delivery methods demonstrated small, but statistically significant improvements in QOL measures. However, there were no statistically significant differences in effectiveness between the telehealth interventions and UC (p = 0.76). The results indicate that telehealth interventions are as effective at improving QOL scores in patients undergoing cancer treatment as in-person UC. Further studies should be undertaken on different modalities of telehealth to determine its appropriate and effective use in interventions to improve the QOL for cancer patients undergoing treatment.


Hippocampal sclerosis revisited.
Studies dating back more than 150 years reported a relationship between hippocampal sclerosis and epilepsy. Retrospective studies of patients who underwent temporal lobectomy for intractable partial epilepsy found a relationship between a history of early childhood convulsions, hippocampal sclerosis, and the development of temporal lobe epilepsy. Many believe that febrile seizures lead to hippocampal damage and this in turn predisposes the patient to the development of temporal lobe epilepsy. Studies in adult rats have shown that seizures can lead to hippocampal damage and unprovoked recurrent seizures. However, many questions remain as to the relevance of early childhood seizures to hippocampal sclerosis and temporal lobe epilepsy. Human prospective epidemiologic studies have not shown a relationship between early childhood seizures and temporal lobe epilepsy. Recent MRI studies in humans suggest that a preexisting hippocampal lesion may predispose infants to experience febrile seizures, later on hippocampal sclerosis, and possibly temporal lobe epilepsy may occur. Unlike the studies in adult rats, normal immature rats with seizures have not been shown to develop hippocampal damage or unprovoked seizures in adulthood. Furthermore, animal studies reveal that preexisting brain abnormalities can predispose to hippocampal damage following seizures early in life. This paper reviews evidence for and against the view that early childhood convulsions, hippocampal sclerosis, and temporal lobe epilepsy are related, while also exploring clinical and animal studies on how seizures can lead to hippocampal damage, and how this can result in temporal lobe epilepsy. By better understanding the cause and effect relationship between early childhood seizures and hippocampal injury in normal and abnormal brains specific treatments can be developed that target the pathogenesis of epilepsy.


[Detection of bdellovibrios and bacteriophages in the seawater near the shores].
The frequency of occurrence of bdellovibrios and intestinal bacteriophages at the sites of contamination in the sea and along the sea shore and at the points remote from the sources of contamination was studied. At the contaminated sites bdellovibrios were revealed in 73.3-100% and bacteriophage in 75-83.4% of the samples. There was found to be a moderate correlative association between the bdellovibrios, bacteriophages and the coli titre. In the water of the shore region of the sea bdellovibrios were found in 15.4%, and bacteriophages in 20.9% of the samples. An attempt at a sanitary assessment of beaches by the data of bdellovibrio detection was unsuccessful. There was found no correlative association between the coli-titre and the bdellovibrios in the water of the shore area. Bdellovibrio proved to play an insignificant role in the processes of self-purification from the intestinal microbial flora of weakly and moderately contaminated sea water. Taking into consideration the high resistance in the sea water of the intestinal bacteriophages preference was given to them over the bdellovibrios as the index of the sea water contamination.


Unpredictable food availability induces metabolic and hormonal changes independent of food intake in a sedentary songbird.
Environments often vary with regard to their temporal resource availability, but little is understood concerning how resource predictability impacts animals. The adaptive regulation hypothesis suggests that organisms act to conserve their current energetic state during periods of diminished food access and recuperate their energetic reserves (fat and muscle) during periods of greater food availability. In contrast, the chronic stress hypothesis suggests that variation in access to food can induce a prolonged stress response, resulting in maladaptive usage of energy reserves and increased behavioral activity. To distinguish between these hypotheses we compared the behavioral, hormonal and metabolic responses of captive curve-billed thrashers, Toxostoma curvirostre, fed varying amounts each day (variable group) with those of birds fed a constant amount every day (constant feeding group). Birds of both groups consumed, on average, a similar total amount of food during the course of the study, but birds in the variable feeding group lost mass and increased their circulating initial levels of the stress hormone corticosterone, showed evidence for increased secretion of a hypothalamic stress peptide, vasotocin, used greater amounts of fat and protein energy reserves, and were more behaviorally active than birds in the constant feeding group. Overall, these findings support the chronic stress hypothesis and suggest that birds such as thrashers may be particularly susceptible to the perception of unpredictable variation in food supplies independent of actual energetic constraints.


All-trans-retinoic acid intensifies endoplasmic reticulum stress in N-acetylglucosaminyltransferase V repressed human hepatocarcinoma cells by perturbing homocysteine metabolism.
We previously reported that all-trans-retinoic acid (ATRA) induced apoptosis in N-acetylglucosaminyltransferase V (GnT-V) repressed human hepatocarcinoma 7721 (GnT-V-AS/7721) cells via endoplasmic reticulum (ER) stress. In addition to confirming these findings, we further found that ATRA repressed the expression of betaine-homocysteine methyltransferase (BHMT) and cystathionine-beta-synthase (CBS), which are key enzymes that are involved in homocysteine metabolism, increased the level of intracellular homocysteine, and decreased the glutathione (GSH) level in GnT-V-AS/7721 cells. To investigate the effect of ATRA on homocysteine metabolism, cells were challenged with exogenous homocysteine. In GnT-V-AS/7721 cells with ATRA treatment, a significant elevation of intracellular homocysteine levels suggests that ATRA perturbs homocysteine metabolism in GnT-V-AS/7721 cells and, therefore, sensitizes the cells to homocysteine-induced ER stress. An obvious increase in the levels of GRP78/Bip protein and spliced XBP1 mRNA were observed. Furthermore, we observed that ATRA blunted the homocysteine-induced increase of GSH only in GnT-V-AS/7721 cells. These results demonstrate that ATRA intensifies ER stress and induces apoptosis in GnT-V-AS/7721 cells by disturbing homocysteine metabolism through the down-regulation of CBS and BHMT, depleting the cellular GSH and, in turn, altering the cellular redox status. In addition, we showed that ATRA did not trigger ER stress, induce apoptosis, or affect homocysteine metabolism in L02 cells, which is a cell type that is derived from normal liver tissue. These results provide support for the hypothesis that ATRA is an anticancer agent.


Medically Graded Honey Supplementation Formula to Preterm Infants as a Prebiotic: A Randomized Controlled Trial.
The aim of the study was to assess the effect of medically graded enteral honey supplementation on the intestinal microbiota, immune response, and somatic growth of preterm infants. A prospective randomized controlled trial was conducted on preterm infants with gestational age ≤34 weeks and postnatal age >3 days. After reaching 1/2 goal enteral feeds, medically graded bee honey was added to milk at a dose of 5, 10, 15, and 0 g/day for 2 weeks in groups A, B, C, and D, respectively. Anthropometric measurements, CD4 and CD8 cytokines, stool cultures, and stool polymerase chain reaction assays for molecular detection of microbiomes were performed at 0, 7, and 14 days of intervention. Analysis of variance test was used to detect differences among the 4 groups. A total of 40 subjects were enrolled; 10 in each arm of the study. Compared with group D, all 3 intervention groups demonstrated significant increase in weight (P < 0.0001). Head circumference increased in groups B and C (P = 0.0056). There were no changes in CD4 or CD8 cytokines (P = 0.24 and P = 0.11, respectively). Enterobacter stool colonization decreased in groups A and B (P = 0.002), whereas Bifidobacterium bifidum colony counts increased in groups A, B, and C (P = 0.002) and lactobacilli colony counts increased in group B (P < 0.0001). Applying real-time polymerase chain reaction, B bifidum and lactobacilli increased in group C (P < 0.0001). Supplementation of milk formula with medically graded honey was associated with changes in physical growth and colonic microbiota of preterm infants. Further studies are needed to examine the sustainability of these effects and associated long-term outcomes.


Mechanisms of Self-Assembly and Energy Harvesting in Tuneable Conjugates of Quantum Dots and Engineered Photovoltaic Proteins.
Photoreaction centers facilitate the solar energy transduction at the heart of photosynthesis and there is increasing interest in their incorporation into biohybrid devices for solar energy conversion, sensing, and other applications. In this work, the self-assembly of conjugates between engineered bacterial reaction centers (RCs) and quantum dots (QDs) that act as a synthetic light harvesting system is described. The interface between protein and QD is provided by a polyhistidine tag that confers a tight and specific binding and defines the geometry of the interaction. Protein engineering that changes the pigment composition of the RC is used to identify Förster resonance energy transfer as the mechanism through which QDs can drive RC photochemistry with a high energy transfer efficiency. A thermodynamic explanation of RC/QD conjugation based on a multiple/independent binding model is provided. It is also demonstrated that the presence of multiple binding sites affects energy coupling not only between RCs and QDs but also among the bound RCs themselves, effects which likely stem from restricted RC dynamics at the QD surface in denser conjugates. These findings are readily transferrable to many other conjugate systems between proteins or combinations of proteins and other nanomaterials.


Elements Selection for Transcostal HIFU Refocusing Method: Simulation Study.
Treatment of tumors in organs obscured by the ribs is a challenge for high-intensity focused ultrasound (HIFU) array. The ribs absorb the ultrasound beam's energy causing the temperature of the ribs to rise, the beam to be distorted, and the focal heat deposition to be limited. The challenges presented by the ribs motivated the development of the limited power deposition (LPD) refocusing algorithm, which is capable of limiting the power deposition over the ribs while maximizing the energy deposition at the focus, thus achieving an efficient and safe ablation. In this article, a new iterative sparse LPD (ISLPD) approach that provides similar focal heating to previously developed methods while reducing the number of transducers excited during the HIFU treatment is discussed. It will be possible to conduct processes, such as imaging and motion tracking in parallel with the HIFU treatment by using the elements no longer used by the refocusing technique. The approach removes transducer elements by using an iteratively reweighted penalty matrix to achieve a more sparse solution over the course of multiple rounds of sparsity induction by harshly punishing the use of elements that previously had low excitation magnitudes. The semidefinite relaxation (SDR) method is used as a means of relaxing the nonconvex constraints into convex form and induces sparsity using the one-norm squared as a convex surrogate for the zero-norm. A 1-MHz spherical phased-array focused on a target in an inhomogeneous medium is simulated to test the algorithms' efficacy. The propagation of the waves as they travel from the array toward the target was modeled using a finite-difference time-domain propagation model. Temperature simulations that utilized the inhomogeneous bioheat transfer equation (BHTE) were used to determine the temperature rise profile within the region of interest (ROI). These simulation results illustrate the benefits of the optimization-based approach proposed in this article over the ray-tracing (shadowing) method for element selection.


[Health education in the care to clients of the blood glucose self-monitoring program].
This article reports, in a systemized and analytical way, the experience of an Outreach Program in the period between 2010 and 2011. The study focused on health education interventions as strategies to improve the adherence of individuals with insulin- dependent diabetes mellitus (IDDM), clients of a blood glucose self-Monitoring program. In addition, we intended to contribute to the reorganization of the program's working processes in the unit. Health education strategies were used in both educational groups and home visits, thus permitting the provision of care that was more individualized. Data regarding the clients were organized on a spreadsheet and in files for the Family Health teams, which made it easier to identify the patients, including those who were absent, helping to decentralize the care. By using health education strategies, we intended to contribute to a more comprehensive and emancipatory care of the clients, aimed at a continuous reflection of the workers regarding their practices.


Diagnosis of irreversible dementia: how extensive the evaluation?
Recommendations for managing the patient with irreversible dementia can be organized according to the results of the initial evaluation. The patient may have normal mental status and normal functional assessment, abnormal mental status and abnormal functional assessment, or mixed results (one normal, one abnormal). The extent of further investigations usually depends on initial clinical findings and on patient and/or family preferences. MRI imaging may be useful prior to referral if greater specificity in diagnosis is desired. Referral is suggested for illnesses of short duration, atypical features, or when the patient and family require precision in diagnostic labeling.


International Measurement Evaluation Programme: IMEP-9, trace elements in water.
The International Measurement Evaluation Programme (IMEP) is an interlaboratory comparison scheme, founded, owned and coordinated by the Institute for Reference Materials and Measurements (IRMM) since 1988. IMEP-9 is the third round of trace elements in water evaluation following IMEP-3 and IMEP-6. Reference values for 15 elements stating total concentrations and combined uncertainties (according to GUM) were established. The reference values were established mainly by isotope dilution mass spectrometry (IDMS) as a primary method of measurement, and values traceable to the SI were obtained. The four elements that could not be certified by IDMS were assigned values by means of other measurement techniques. Results from 201 laboratories from 35 countries and four continents were evaluated against the reference values and the comparability between the laboratories is presented graphically.


Molecular pathogenesis of progression to myeloid leukemia from TET-insufficient status.
Loss-of-function mutations in ten-eleven translocation-2 (TET2) are recurrent events in acute myeloid leukemia (AML) as well as in preleukemic hematopoietic stem cells (HSCs) of age-related clonal hematopoiesis. TET3 mutations are infrequent in AML, but the level of TET3 expression in HSCs has been found to decline with age. We examined the impact of gradual decrease of TET function in AML development by generating mice with Tet deficiency at various degrees. Tet2f/f and Tet3f/f mice were crossed with mice expressing Mx1-Cre to generate Tet2f/wtTet3f/fMx-Cre+ (T2ΔT3), Tet2f/fTet3f/wtMx-Cre+ (ΔT2T3), and Tet2f/fTet3f/fMx-Cre+ (ΔT2ΔT3) mice. All ΔT2ΔT3 mice died of aggressive AML at a median survival of 10.7 weeks. By comparison, T2ΔT3 and ΔT2T3 mice developed AML at longer latencies, with a median survival of ∼27 weeks. Remarkably, all 9 T2ΔT3 and 8 ΔT2T3 mice with AML showed inactivation of the remaining nontargeted Tet2 or Tet3 allele, respectively, owing to exonic loss in either gene or stop-gain mutations in Tet3. Recurrent mutations other than Tet3 were not noted in any mice by whole-exome sequencing. Spontaneous inactivation of residual Tet2 or Tet3 alleles is a recurrent genetic event during the development of AML with Tet insufficiency.


Arg-72 of pharaonis phoborhodopsin (sensory rhodopsin II) is important for the maintenance of the protein structure in the solubilized states.
In bacteriorhodopsin (bR), Arg-82bR has been proven to be a very important residue for functional role of this light-driven proton pump. The arginine residue at this position is a super-conserved residue among archaeal rhodopsins. pharaonis phoborhodopsin (ppR; or called as "pharaonis sensory rhodopsin II") has its absorption maximum at 498 nm and acts as a sensor in the membrane of Natronobacterium pharaonis, mediating the negative phototaxis from the light of wavelength shorter than 520 nm. To investigate the role of the arginine residue (Arg-72ppR) of ppR corresponding to Arg-82bR, mutants whose Arg-72ppR was replaced by alanine (R72A), lysine (R72K), glutamine (R72Q) and serine (R72S) were prepared. These mutants were unstable in low concentrations of NaCl and lost their color gradually when the proteins were solubilized with 0.1% n-dodecyl-beta-D-maltoside. The order of instability was R72S > R72A > R72K > R72Q > the wild type. The rates of denaturation were reduced in a solution of high concentrations of monovalent anions.


Evaluation of a differential mobility spectrometer/miniature mass spectrometer system.
A planar differential mobility spectrometer (DMS) was coupled to a Mini 10 handheld rectilinear ion trap (RIT) mass spectrometer (MS) (total weight 10 kg), and the performance of the instrument was evaluated using illicit drug analysis. Coupling of DMS (which requires a continuous flow of drift gas) with a miniature MS (which operates best using sample introduction via a discontinuous atmospheric pressure interface, DAPI), was achieved with auxiliary pumping using a 5 L/min miniature diaphragm sample pump placed between the two devices. On-line ion mobility filtering showed to be advantageous in reducing the background chemical noise in the analysis of the psychotropic drug diazepam in urine using nanoelectrospray ionization. The combination of a miniature mass spectrometer with simple and rapid gas-phase ion separation by DMS allowed the characteristic fragmentation pattern of diazepam to be distinguished in a simple urine extract at lower limits of detection (50 ng/mL) than that achieved without DMS (200 ng/mL). The additional separation power of DMS facilitated the identification of two drugs of similar molecular weight, morphine (average MW = 285.34) and diazepam (average MW = 284.70), using a miniature mass spectrometer capable of unit resolution. The similarity in the proton affinities of these two compounds resulted in some cross-interference in the MS data due to facile ionization of the neutral form of the compound even when the ionic form had been separated by DMS.


Body Mass Index and Associated Clinical Variables in Patients with Non-Celiac Wheat Sensitivity.
Non-Celiac Wheat Sensitivity (NCWS) is still a largely undefined condition, due to the lack of a diagnostic marker. Few data are available about the nutritional characteristics of NCWS patients at diagnosis. To evaluate the proportion of NCWS patients who were underweight, normal weight, overweight, or obese at diagnosis, and to search for possible correlations between their Body Mass Index (BMI) and other NCWS-related disease characteristics. The clinical charts of 145 NCWS patients (125 F, 20 M, mean age 37.1 ± 11.4 years), diagnosed between January 2012 and March 2018, were reviewed. As a comparison, 84 celiac disease (CD) patients (73 F, 11 M, mean age 39.8 ± 13.9 years) were evaluated. All NCWS diagnoses were based on a double-blind placebo-controlled wheat challenge (DBPCWC) method. BMI distribution was similar in the NCWS (6.2% underweight and 15.2% obese subjects) and CD patients (6% underweight and 7.1% obese subjects). Underweight NCWS subjects were significantly younger and had a shorter clinical history than the overweight or obese ones. Unlike the other NCWS patients, none of them had a DQ2 and/or DQ8 haplotype. Overweight and obese NCWS patients were more frequently suffering from associated autoimmune diseases than the other BMI categories (P = 0.05). Compared to the CD controls, NCWS patients showed a higher frequency of Irritable Bowel Syndrome (IBS)-like (P = 0.01) and extraintestinal symptoms (P = 0.03) and a longer clinical history (P = 0.04), whereas weight loss was more frequent in CD (P = 0.02). NCWS patients showed a BMI distribution similar to CD patients. However, NCWS was found to be a heterogenous condition that regards BMI, and clinical characteristics differed between the underweight and overweight/obese patients.


Paclitaxel-eluting silicone airway stent for preventing granulation tissue growth and lung cancer relapse in central airway pathologies.
Airway stents are used to treat obstructive central airway pathologies including palliation of lung cancer, but face challenges with granulation tissue growth. Paclitaxel is a chemotherapy drug that also suppresses growth of granulation tissue. Yet, side effects arise from administration with toxic solubilizers. By incorporating paclitaxel in silicone stents, delivery of paclitaxel can be localized, and side effects minimized. Paclitaxel was incorporated into Liquid Silicone Rubber (LSR) containing polydimethylsiloxane, either as a powder or solution, prior to curing. Drug release study was compared in vitro at 37°C over 10 days. Drug release was quantified using HPLC, and bronchial cell lines were grown on LSR to investigate drug cytotoxicity, and expression of inflammatory markers, specifically interleukin-6 and interleukin-8. Release rate of paclitaxel incorporated into silicone rubber was consistent with the Korsmeyer and Weibull models (R2 > 0.96). Paclitaxel exposure reduced IL-8 levels in cancer cell lines, whilst no cytotoxic effect was observed in all cell lines at treatment concentration levels (≤ 0.1% (w/v) paclitaxel in silicone). Incorporating paclitaxel into a silicone matrix for future use in a tracheobronchial stent was investigated. Drug release from silicone was observed and is a promising avenue for future treatments of central airway pathologies.


A junction branch point adjacent to a DNA backbone nick directs substrate cleavage by Saccharomyces cerevisiae Mus81-Mms4.
The DNA structure-selective endonuclease Mus81-Mms4/Eme1 incises a number of nicked joint molecule substrates in vitro. 3'-flaps are an excellent in vitro substrate for Mus81-Mms4/Eme1. Mutants in MUS81 are synthetically lethal with mutations in the 5'-flap endonuclease FEN1/Rad27 in Saccharomyces cerevisiae and Schizosaccharomyces pombe. Considering the possibility for isoenergetic interconversion between 3'- and 5'- flaps, these data are consistent with the hypothesis that Mus81-Mms4/Eme1 acts on 3'-flaps in vivo. FEN1/Rad27 prefers dually flapped substrates and cleaves in a way that allows direct ligation of the resulting nick in the product duplex. Here we test the activity of Mus81-Mms4 on dually flapped substrates and find that in contrast to FEN1/Rad27, Mus81-Mms4 activity is impaired on such substrates, resulting in cleavage products that do not allow direct religation. We conclude that Mus81-Mms4, unlike FEN1/Rad27, does not prefer dually flapped substrates and is unlikely to function as a 3'-flapase counterpart to the 5'-flapase activity of FEN1/Rad27. We further find that joint molecule incision by Mus81-Mms4 occurs in a fashion determined by the branch point, regardless of the position of an upstream duplex end. These findings underscore the significance of a nick adjacent to a branch point for Mus81-Mms4 incision.


Lessons from Hawaii's health care system.
Hawaii has 20 years of experience with employer mandates and universal access. With the U.S. Congress considering the controversial issue of whether to require companies to provide health insurance for employees, Hawaii's approach to health care has some valuable lessons.


Intradermal cell transplantation in soluble collagen.
Intradermal, as opposed to subcutaneous, cell transplantation was previously shown to be advantageous for tumor cell growth, but this site has not been used for transplantation of normal nonneoplastic cells. In preliminary experiments we found that it was difficult to control the size and shape of transplants when we injected dissociated cells intradermally. This problem was solved by placing cells in nongelled, pepsin-solubilized collagen prior to injection. This technique permitted the successful transplantation of normal bovine adrenocortical cells and of neoplastic cells (3T3 cells secreting FGF) in scid mice. Primary bovine adrenocortical cells formed functional vascularized tissue and the transplants rescued the animals from the lethal effects of adrenalectomy. The histological structure of transplant tissues resembled that previously observed when cells were transplanted in the subrenal capsule space. We also used a line of 3T3 cells that has been genetically modified to secrete a form of acidic FGF. When transplanted intradermally in collagen, they formed rapidly enlarging masses of cells that could easily be palpated beneath the skin of the animal. Intradermal injection of cells in pepsin-solubilized collagen is a simple and reliable technique for transplanting normal primary cells and preneoplastic cells. The ability to grow both types of cells in an easily accessible site allows less invasive monitoring of growth, angiogenesis, and other features of the transplant.


Yeast homologs of human MCUR1 regulate mitochondrial proline metabolism.
Mitochondria house evolutionarily conserved pathways of carbon and nitrogen metabolism that drive cellular energy production. Mitochondrial bioenergetics is regulated by calcium uptake through the mitochondrial calcium uniporter (MCU), a multi-protein complex whose assembly in the inner mitochondrial membrane is facilitated by the scaffold factor MCUR1. Intriguingly, many fungi that lack MCU contain MCUR1 homologs, suggesting alternate functions. Herein, we characterize Saccharomyces cerevisiae homologs Put6 and Put7 of MCUR1 as regulators of mitochondrial proline metabolism. Put6 and Put7 are tethered to the inner mitochondrial membrane in a large hetero-oligomeric complex, whose abundance is regulated by proline. Loss of this complex perturbs mitochondrial proline homeostasis and cellular redox balance. Yeast cells lacking either Put6 or Put7 exhibit a pronounced defect in proline utilization, which can be corrected by the heterologous expression of human MCUR1. Our work uncovers an unexpected role of MCUR1 homologs in mitochondrial proline metabolism.


Pancreatic islet transplantation.
Type 1 diabetes mellitus is an autoimmune disease, which results in the permanent destruction of β-cells of the pancreatic islets of Langerhans. While exogenous insulin therapy has dramatically improved the quality of life, chronic diabetic complications develop in a substantial proportion of subjects and these complications generally progress and worsen over time. Although intensive insulin therapy has proven effective to delay and sometimes prevent the progression of complications such as nephropathy, neuropathy or retinopathy, it is difficult to achieve and maintain long term in most subjects. Reasons for this difficulty include compliance issues and the increased risk of severe hypoglycemic episodes, which are generally associated with intensification of exogenous insulin therapy. Clinical studies have shown that transplantation of pancreas or purified pancreatic islets can support glucose homeostasis in type 1 diabetic patients. Islet transplantation carries the special advantages of being less invasive and resulting in fewer complications compared with the traditional pancreas or pancreas-kidney transplantation. However, islet transplantation efforts have limitations including the short supply of donor pancreata, the paucity of experienced islet isolation teams, side effects of immunosuppressants and poor long-term results. The purpose of this article is to review recent progress in clinical islet transplantation for the treatment of diabetes.


COVID-19-related premature ovarian insufficiency: case report and literature review.
The aim of this study is to present the case report of a 36-year-old woman developing premature ovarian insufficiency (POI) after COVID-19 and review the literature referring to the possible impact of SARS-CoV-2 infection on female reproduction. A 36-year-old nulligravida with normal menstrual cycles, non-smoker, with a normal body mass index and no pelvic surgery or oncological treatment in her medical history presented to the Infertility Center of the Institute of Mother and Child in Warsaw after a year of unsuccessful attempts to get pregnant. During diagnostic process she was affected by COVID-19 with a mild manifestation and thereafter she presented amenorrhea with intense hot flushes. Further diagnostic confirmed the diagnosis of POI. There is a strong molecular basis for a possible effect of SARS-CoV-2 infection on the female reproductive system; however, the results of available research are conflicting. All of these aspects are discussed in detail. SARS-CoV-2 infection may cause serious complications that cast a long shadow on a patient's future life and health. Further research is needed to assess the real impact of SARS-CoV-2 infection on female reproductive health, as well as potential preventive and therapeutic strategies for women affected with COVID-19.


Stresses exerted in the hindlimb muscles of common chimpanzees (Pan troglodytes) during bipedal locomotion.
Recent studies have indicated that chimpanzee bipedality is mechanically inefficient and dynamically unlike that of humans, thus undermining the chimpanzee analogy for mechanical aspects of the early evolution of hominid bipedalism. This paper continues this theme by measuring the forces and stresses engendered by the muscles during bipedal locomotion, for an untrained chimpanzee and for data from chimpanzees which have been encouraged to walk bipedally, presented in the literature. Peak stresses in the triceps surae were lower for the untrained chimpanzee than for the trained subjects because during the late stance phase, when peak ankle moments occur, the centre of pressure of the ground reaction force on the foot of the untrained chimpanzee stayed close to the ankle joint. In contrast, for the trained subjects it moved closer to the toes, as in human bipedalism. Quadriceps and hip extensor stresses are approximately 30% larger for the untrained chimpanzee than for the trained subjects, because the trained chimpanzees walked with a more erect posture. These results may reflect the way in which muscles can develop in response to training, since research on humans has shown that muscle physiological cross-sectional area increases as a result of exercise, resulting in smaller stresses for a given muscle force. During a slow walk, untrained chimpanzees were found to exert far greater muscle stresses than humans do when running at moderate speed, particularly in the muscles that extend the hip, because of the bent-hip, bent-knee posture.


[Innovations in the treatment of gastroduodenal ulcers].
It has long been known that gastroduodenal ulcers may heal spontaneously; this is the placebo effect in which the healing rate ranges from 20% at 2 weeks to 80% at 8 weeks. Two or three weeks can be gained by using cimetidine or ranitidine, with healing rates of 42% at 2 weeks and 85% at 6 weeks. Ranitidine probably has less side-effects than cimetidine. A new drug, omeprozole, appears to be much more potent but is not yet available. A vagolytic drug with specific affinity to the stomach (pirenzepine), as well as low-dosage bismuth bi-citrate and gastric mucosa protectors (sucralfate) are now returning in force.


[Department of Protein Structure and Function--Volodimir Oleksandrovich Bielitser School of the Ukrainian Academy of Sciences. Studies on structure of fibrin fiber structure and mechanism (1975-1987].
In this short historical review the records about foundation and research activity of the Department of Structure and Function of Protein--school of V. A. Belitser, Member of the National Academy of Sciences of Ukraine are presented. V. A. Belitser was the founder and indispensable chief of the department since the date of its creation (1944) till 1987. The main research interests (1975-1987) of the department were focused at the investigation of structure, biological function of the fibrinogen-fibrin system, mechanisms of the network assembly and of the fibrin fibers structure. Studying the molecular mechanisms of the fibrin fiber assembly, it was shown that the specificity of the building structure was shown is determined by the specific reactive sites with strong affinity of the molecules. The activity of the sites was investigated on protein molecules as well as the fragments. The physical nature of the bonds created by the active sites, that appearing during in the process of fibrinogen activation by thrombin, was revealed. Examination of the fibrin assembly in cooperation with electronmicroscopists and studies of the complex formation between active fragments and fibrin monomer were summarized. Both the fibrin monomer polymerization and protofibril lateral association are presented as two stages in the assembly of the fibrin network. In the research of the domain fibrinogen structure the specific sites of the fibrin assembly in each of the domains were found. COOH-terminal regions of the A alpha-chains play independent part in the fibrinogen and fibrin. That is why it is relevant to consider them as alpha C-domains. In the free fibrinogen molecules (in solution) these domains are responsible for globular shape, they are linked to domains D intramolecularly. When fibrin assembly takes place, alpha C-domains play significant carriage role in fibrin molecules interaction, linking to domains D intermolecularly. The model of the fibrinogen molecule structure and the general scheme of the fibrin fibers network formation were proposed. Physico-chemical basics of a biological structure assembly were elucidated using the process of the fibrin self-assembly as an example. Much attention was devoted to the problems of practical medicine. The quantitative methods of fibrinogen, soluble fibrin and active fibrin/fibrinogen fragments estimation in blood plasma were developed.


60Co gamma-rays induce predominantly C/G to G/C transversions in double-stranded M13 DNA.
Upon irradiation with gamma rays of an oxygenated aqueous solution of double-stranded M13 DNA, a very specific mutation spectrum was found with respect to both the type and the positions in the DNA sequence. Of the 23 mutations, which were sequenced, 16 represent a C/G to G/C transversion. A C/G to T/A transition was found once and a G/C to T/A transversion twice. The remaining 4 mutations are frameshifts, 2 are identical and formed by the insertion of a G/C basepair; the other 2 mutations are due to a duplication of 10 basepairs situated at different positions but with a remarkable homology in base sequence. Fourteen mutations, including the 2 duplications are found in the neighbourhood of a TGCT/ACGA sequence.


Bilateral sixth nerve pareses with temporal arteritis and diabetes.
A 78-year-old white woman presented with a 1-month history of neck and right shoulder pain and a 12-day history of acute horizontal diplopia. On examination, bilateral sixth nerve pareses were present, right greater than left. On evaluation, the patient was noted to have normal fasting blood sugars, but moderately elevated blood sugars after a glucose load. A temporal artery biopsy was performed which was positive. Whether diabetic or arteritic, bilateral sixth nerve pareses resulted in our patient seeking medical attention. By considering temporal arteritis, it is possible that blindness was avoided.


NADK2 positively modulates abscisic acid-induced stomatal closure by affecting accumulation of H2O2, Ca2+ and nitric oxide in Arabidopsis guard cells.
NAD kinase2 (NADK2) plays key roles in chloroplastic NADP biosynthesis, stress adaptation and modulation of cellular metabolisms in Arabidopsis. However, it is unknown whether and how NADK2 affects abscisic acid (ABA)-mediated stomatal movement. Here, we detected that null mutant nadk2 was more sensitive to drought stress than WT, and NADK2 gene was active in guard cells. Furthermore, NADK2 mutation impaired ABA-induced stomatal closure and ABA inhibition of light-promoted stomatal opening. NADK2 disruption also impaired ABA-stimulated accumulation of H2O2, Ca2+ and nitric oxide (NO) in guard cells, but did not affect the stomatal closure evoked by exogenous H2O2, Ca2+ or NO. Expression analysis revealed that ABA-promoted increases in transcripts of AtrbohD, AtrbohF and NIA1 were markedly arrested in guard cells of nadk2 compared with those of WT. Besides, genetic evidence indicated that NADK2 acted synergistically with OST1 and ABI1 during ABA-induced stomatal closure. Together, these results suggest that NADK2 is an essential positive regulator, and functions upstream of H2O2 in guard cell ABA signaling. It stimulates stomatal closure mainly through increasing the generation of H2O2, Ca2+ and NO in guard cells in Arabidopsis.


In vivo protein turnover rates in varying oxygen tensions nominate MYBBP1A as a mediator of the hyperoxia response.
Oxygen deprivation and excess are both toxic. Thus, the body's ability to adapt to varying oxygen tensions is critical for survival. While the hypoxia transcriptional response has been well studied, the post-translational effects of oxygen have been underexplored. In this study, we systematically investigate protein turnover rates in mouse heart, lung, and brain under different inhaled oxygen tensions. We find that the lung proteome is the most responsive to varying oxygen tensions. In particular, several extracellular matrix (ECM) proteins are stabilized in the lung under both hypoxia and hyperoxia. Furthermore, we show that complex 1 of the electron transport chain is destabilized in hyperoxia, in accordance with the exacerbation of associated disease models by hyperoxia and rescue by hypoxia. Moreover, we nominate MYBBP1A as a hyperoxia transcriptional regulator, particularly in the context of rRNA homeostasis. Overall, our study highlights the importance of varying oxygen tensions on protein turnover rates and identifies tissue-specific mediators of oxygen-dependent responses.


Evolutionary development of tensegrity structures.
Contributions from the emerging fields of molecular genetics and evo-devo (evolutionary developmental biology) are greatly benefiting the field of evolutionary computation, initiating a promise of renewal in the traditional methodology. While direct encoding has constituted a dominant paradigm, indirect ways to encode the solutions have been reported, yet little attention has been paid to the benefits of the proposed methods to real problems. In this work, we study the biological properties that emerge by means of using indirect encodings in the context of form-finding problems. A novel indirect encoding model for artificial development has been defined and applied to an engineering structural-design problem, specifically to the discovery of tensegrity structures. This model has been compared with a direct encoding scheme. While the direct encoding performs similarly well to the proposed method, indirect-based results typically outperform the direct-based results in aspects not directly linked to the nature of the problem itself, but to the emergence of properties found in biological organisms, like organicity, generalization capacity, or modularity aspects which are highly valuable in engineering.


Localization of smooth muscle and nonmuscle actin isoforms in the human aqueous outflow pathway.
alpha-Smooth muscle actin is the isoform of actin restricted to vascular smooth muscle, pericytes, myofibroblasts and, certain other cells that are of myoid origin. We investigated the distribution of alpha-smooth muscle actin and nonmuscle specific filamentous actin in the human aqueous outflow system by immunohistochemical methods. Filamentous actin was observed in all cellular constituents of the outflow pathway, while distribution of alpha-smooth muscle actin was restricted to the ciliary muscle, to specific cells throughout the trabecular meshwork, and to cells adjacent to the outer wall and the collector channels. The ciliary muscle extended deep into the corneoscleral meshwork, far anterior to the scleral spur. These findings agree with our previous study localizing the distribution of smooth muscle myosin in the human aqueous outflow pathway. Although functionality of the immunoreactive cells needs to be demonstrated, our data show that a potentially contractile apparatus exists in a subpopulation of trabecular meshwork cells and in certain cells of the more distal components of the outflow system.


[Influence of hepatitis C virus replication on splenic lymphoma with villous lymphocytes].
The association between hepatitis C and B-cell non-Hodgkin lymphoma (NHL) has been suggested by several studies. We report the case of a 70 year-old patient with the diagnosis of chronic hepatitis C and splenic lymphoma with villous lymphocytes, who had undergone splenectomy and chemotherapy with fludarabine, with transient effectiveness. He was sent to our Hepatology Clinic for the treatment of hepatitis C, and Pegylated Interferon and Ribavirin were started. He had virological and hematological response (63% of villous lymphocytes in the peripheral blood at the beginning of therapy and 0% at the end). With the suspension of antiviral therapy, recurrence of HCV infection and reappearance of atypical lymphocytes (24%) were observed, and Pegylated Interferon was restarted, with good response. The relationship between viral and hematologic response (remission and relapse) supports the hypothesis that hepatitis C virus has an active role in the pathogenesis of splenic lymphoma with villous lymphocytes.


Management of major organ involvement of Behçet's syndrome: a systematic review for update of the EULAR recommendations.
To assess the efficacy and safety of treatment modalities for major organ involvement of Behçet's syndrome (BS), in order to inform the update of the EULAR recommendations for the management of BS. A systematic literature review of all randomized controlled trials, controlled clinical trials, or open label trials assessing eye, vascular, nervous system or gastrointestinal system involvement of BS was performed. If controlled trials were not available for answering a specific research question, uncontrolled studies or case series were also included. We reviewed the titles and abstracts of 3927 references and 161 studies met our inclusion criteria. There were only nine randomized controlled trials. Observational studies with IFN-α and monoclonal anti-TNF antibodies showed beneficial results for refractory uveitis. Meta-analysis of case-control studies showed that immunosuppressives decreased the recurrence rate of deep vein thrombosis significantly whereas anticoagulants did not. CYC and high dose glucocorticoids decreased mortality in pulmonary arterial aneurysms and postoperative complications in peripheral artery aneurysms. Beneficial results for gastrointestinal involvement were obtained with 5-ASA derivatives and AZA as first line treatment and with thalidomide and/or monoclonal anti-TNF antibodies in refractory cases. Observational studies for nervous system involvement showed improved outcome with immunosuppressives and glucocorticoids. Meta-analysis of case-control studies showed an increased risk of developing nervous system involvement with ciclosporin-A. The majority of studies related to major organ involvement that informed the updated EULAR recommendations for the management of BS were observational studies.


Regulator of G-Protein Signalling-14 (RGS14) Regulates the Activation of αMβ2 Integrin during Phagocytosis.
Integrin-mediated phagocytosis, an important physiological activity undertaken by professional phagocytes, requires bidirectional signalling to/from αMβ2 integrin and involves Rap1 and Rho GTPases. The action of Rap1 and the cytoskeletal protein talin in activating αMβ2 integrins, in a RIAM-independent manner, has been previously shown to be critical during phagocytosis in mammalian phagocytes. However, the events downstream of Rap1 are not clearly understood. Our data demonstrate that one potential Rap1 effector, Regulator of G-Protein Signalling-14 (RGS14), is involved in activating αMβ2. Exogenous expression of RGS14 in COS-7 cells expressing αMβ2 results in increased binding of C3bi-opsonised sheep red blood cells. Consistent with this, knock-down of RGS14 in J774.A1 macrophages results in decreased association with C3bi-opsonised sheep red blood cells. Regulation of αMβ2 function occurs through the R333 residue of the RGS14 Ras/Rap binding domain (RBD) and the F754 residue of β2, residues previously shown to be involved in binding of H-Ras and talin1 head binding prior to αMβ2 activation, respectively. Surprisingly, overexpression of talin2 or RAPL had no effect on αMβ2 regulation. Our results establish for the first time a role for RGS14 in the mechanism of Rap1/talin1 activation of αMβ2 during phagocytosis.


Monocytic differentiation of leukemic HL-60 cells induced by co-treatment with TNF-alpha and MK886 requires activation of pro-apoptotic machinery.
The block of hematopoietic differentiation program in acute myeloid leukemia cells can be overcome by differentiating agent like retinoic acid, but it has several side effects. A study of other differentiation signaling pathways is therefore useful to predict potential targets of anti-leukemic therapy. We demonstrated previously that the co-treatment of HL-60 cells with Tumor necrosis factor-alpha (TNF-alpha) (1 ng/mL) and inhibitor of 5-lipoxygenase MK886 (5 microm) potentiated both monocytic differentiation and apoptosis. In this study, we detected enhanced activation of three main types of mitogen-activated protein kinases (MAPKs) (p38, c-Jun amino-terminal kinase [JNK], extracellular signal-regulated kinase [ERK]), so we assessed their role in differentiation using appropriate pharmacologic inhibitors. The inhibition of pro-apoptotic MAPKs (p38 and JNK) suppressed the effect of MK886 + TNF-alpha co-treatment. On the other hand, down-regulation of pro-survival ERK pathway led to increased differentiation. Those effects were accompanied by increased activation of caspases in cells treated by MK886 + TNF-alpha. Pan-caspase inhibitor ZVAD-fmk significantly decreased both number of apoptotic and differentiated cells. The same effect was observed after inhibition of caspase 9, but not caspase 3 and 8. To conclude, we evidenced that the activation of apoptotic processes and pathways supporting apoptosis (p38 and JNK MAPKs) is required for the monocytic differentiation of HL-60 cells.


Prolonged hypotension following elective stenting of an internal carotid artery stenosis.
We report a case of a 72-year-old man who had undergone elective stenting of an asymptomatic high grade stenosis of the proximal left internal carotid artery. The indication was progression of the stenosis over a 3 year period despite receiving maximal medical therapy, and patient preference for stenting over endarterectomy. He became profoundly hypotensive in the immediate post-stent period and required immediate and prolonged vasopressor support over the next week. The etiological hypothesis is baroreceptor hyperactivity related to procedure induced unilateral stretching of the carotid body.


Clinical features and natural history of intramedullary spinal cord metastasis.
Five cases of intramedullary spinal cord metastasis are presented and an additional 50 cases from the English language literature since 1960 are reviewed. Lung cancer and breast cancer were the most frequently occurring primary neoplasms, but a wide variety of solid tumors may cause intramedullary spinal cord metastasis. The presenting symptoms were pain and/or weakness. The neurologic status deteriorated rapidly in the majority of patients in a period to days to weeks. Progression to a cord hemisection syndrome or cord transection occurred in approximately half of the patients. The characteristic myelographic appearance of fusiform swelling of the cord was seen in one third of the patients, but the myelogram was normal in 42%. Plain radiographs of the spine showed no evidence of metastatic disease in three fourths of cases. The cerebrospinal fluid protein level was frequently elevated, but results of cytologic studies were usually negative. High-resolution computer-assisted tomographic scanning may show intramedullary metastases. Radiation therapy combined with corticosteroid administration offers the only effective palliation. The recognition of intramedullary spinal cord metastasis is an ominous finding. Intramedullary spinal cord metastasis generally occurred in the setting of widespread systemic and intracranial disease, but occasionally was the only site of relapse. More than 80% of patients died within 3 months. Heightened awareness of this entity may lead to early diagnosis at a stage when neurologic deficits are reversible and, it is hoped, more effective palliation can be achieved.


Diarylcoumarins inhibit mycolic acid biosynthesis and kill Mycobacterium tuberculosis by targeting FadD32.
Infection with the bacterial pathogen Mycobacterium tuberculosis imposes an enormous burden on global public health. New antibiotics are urgently needed to combat the global tuberculosis pandemic; however, the development of new small molecules is hindered by a lack of validated drug targets. Here, we describe the identification of a 4,6-diaryl-5,7-dimethyl coumarin series that kills M. tuberculosis by inhibiting fatty acid degradation protein D32 (FadD32), an enzyme that is required for biosynthesis of cell-wall mycolic acids. These substituted coumarin inhibitors directly inhibit the acyl-acyl carrier protein synthetase activity of FadD32. They effectively block bacterial replication both in vitro and in animal models of tuberculosis, validating FadD32 as a target for antibiotic development that works in the same pathway as the established antibiotic isoniazid. Targeting new steps in well-validated biosynthetic pathways in antitubercular therapy is a powerful strategy that removes much of the usual uncertainty surrounding new targets and in vivo clinical efficacy, while circumventing existing resistance to established targets.


Antiviral activities of compounds from aerial parts of Salvia plebeia R. Br.
Salvia plebeia R. Br. is an edible plant widely spread in many countries. It has been used as a traditional medicine to treat common cold, flu, cough, hepatitis, hemorrhoids, etc. The purpose of the study is to explicate antiviral compounds responsible for its traditional use for the common cold or flu. The methanolic extract of the aerial parts of S. plebeia was extracted with CHCl3, EtOAc, and n-BuOH, successively. The EtOAc and CHCl3 fractions were subjected to a successive of chromatographic method, which led to the isolation of fourteen compounds. Inhibition activities of the isolated compounds were evaluated against influenza A (H1N1) neuraminidase. Chemical investigation of the methanolic extracts of S. plebeia resulted in the isolation of two novel benzoylated monoterpene glycosides, named as plebeiosides A (1) and B (2), together with twelve known compounds including four flavonoids (4-5, 7, 10), two sesquiterpenoids (8, 12), four phenolics (9-10, 13-14), a steroid (6), and a triterpenoid (3). Their chemical structures were elucidated based on spectroscopic data and absolute stereochemistries of 1 and 2 were determined by comparison of optical rotations of their hydrolysates with literature values. Compounds 5, 7, 9, and 11 exhibited potent enzymatic inhibition against H1N1 neuraminidase (IC50 values ranging from 11.18±1.73 to 19.83±2.28μM). Furthermore, two flavonoids (5 and 7) and one rosmarinic acid methyl ester (9) reduced cytopathic effects of the H1N1 virus during replication. The antiviral activities of the flavonoids and phenolics isolated from the extracts of S. plebeia supported the traditional application of this medicine on common cold or flu. In this study, benzoylated monoterpene glycosides were first found to exist in this species. Moreover, the present study suggested potential of three compounds (5, 7, and 9) to be new lead structures for the development of new neuraminidase inhibitors in the future.


Effect of caseinate salt addition on the structural characteristics of kefiran systems.
Sodium caseinates-kefiran systems were studied to explore whether any potential interactions between them might exist. The study was performed using low-deformation rheological techniques, which were dynamic and creep tests. The systems were prepared under various experimental conditions such as heating and acidification. Besides, the structure development of the systems in relation to time was also monitored using oscillatory shear rheometry. The results indicated that the structural characteristics of the systems were mainly affected by the state of the caseinates such as the formation of aggregates and to a lesser degree by the interactions of kefiran molecules with the caseinates. Freeze-thaw treatment produced cryogels with good thermal stability and fairly satisfactory mechanical properties. The morphology of the caseinate-kefiran systems was also investigated by means of confocal laser scanning microscopy.


Antioxidant compounds from the stem bark of Garcinia atroviridis.
A new xanthone, namely garcinexanthone G (1), along with eight known compounds, stigmasta-5,22-dien-3β-ol (2), stigmasta-5,22-dien-3-O-β-glucopyranoside (3), 3β-acetoxy-11α,12α-epoxyoleanan-28,13β-olide (4), 2,6-dimethoxy-p-benzoquinone (5), 1,3,5-trihydroxy-2-methoxyxanthone (6), 1,3,7-trihydroxyxanthone (7), kaempferol (8) and quercetin (9), were isolated from the stem bark of Garcinia atroviridis. Their structures were elucidated based on spectroscopic methods including nuclear magnetic resonance (NMR-1D and 2D), UV, IR, and mass spectrometry. All the isolated compounds were evaluated for their antioxidant properties based on the DPPH radical scavenging activities. Results showed that 1,3,7-trihydroxyxanthone and quercetin showed significant antioxidant activities with EC50 values of 16.20 and 12.68 μg/ml, respectively, as compared to the control, ascorbic acid (7.4 μg/ml).


Spontaneous protoplast formation in Methanobacterium bryantii.
Methanobacterium bryantii was found to undergo rapid lysis when grown in a prereduced chemically defined medium under H2-CO2 (4:1, vol/vol). The addition of 20 mM MgCl2 to the medium gave, rather than rapid lysis, a gradual formation of phase-dark spherical bodies which in thin section appeared as true protoplasts. In general, the protoplasts were stabilized by divalent but not monovalent cations and, unlike whole cells, were sensitive to lysis by Triton X-100. Electron microscopic examination revealed that protoplast formation was preceded by a general breakdown of the cell wall with an apparent squeezing out of the protoplast through the degraded wall. The growth of cells was greatly increased and not accompanied by detectable lysis in a medium modified by elevating the levels of nickel and ammonium.


PERsuading nuclear receptors to dance the circadian rhythm.
The recurring light/dark cycle that has a period length of about 24 hours has been internalized in various organisms in the form of a circadian clock. This clock allows a precise orchestration of biochemical and physiological processes in the body thus improving performance. Recently, we found that the clock component PERIOD2 (PER2) can coordinate transcriptional regulation of metabolic, physiological, or behavioral pathways by interacting with nuclear receptors. PER2 appears to act as co-regulator of nuclear receptors linking clock function and transcriptional regulation at the level of protein-protein interactions. Here, we provide additional evidence for modulation of nuclear receptor dependent transcription by PER2 underscoring the broad implication of our finding. Taken together, our findings provide a base for the understanding of various disorders including mood disorders that have their roots in a temporal deregulation of basic metabolic processes.


[Towards the Files. Psychoanalysis and its publishing house strategies].
The following case study deals with the Internationaler Psychoanalytischer Verlag, founded 1919 in Vienna by a group around Sigmund Freud and shut down in 1938 by the Gestapo. During that time the Verlag published the titles of the contemporary psychoanalytic movement, including the first psychoanalytic dictionary, the Almanach as a yearbook, the four authoritative journals, as well as the first complete edition of Freud's writings. A single publishing house became thus responsible for the appearance of an entire theory--a unique situation without historical comparison. My thesis here is that the Verlag initiated certain strategies of publicising (e.g. centralising the movement, enforcing the company's name, labelling its activities) that were, prior to arguments and contents, constitutive for the development of psychoanalysis as well as its implementation within the field of science. Consequently, these strategies cannot be found through interpretations of psychoanalytical texts, but in the genuine files and in view of the material products (books, journals, blurbs, advertisements etc.) that were passed down from the Verlag. As an opening step, this essay explores several of those Viennese files, showing that the Verlag not only struggled with monetary and personal problems but rather and foremost launched branding, marketing and public relation campaigns as key concepts to make sciences and its particular knowledge acceptable. As a result the Verlag changed from a commercial and distributive institution to an epistemic medium.


What is a laser and how is it applied for therapy?
Lasers are now established as a major contributor to modern treatment techniques in a variety of disciplines. They have reached this position by providing improved precision, significantly less invasive surgical procedures and, in some cases, unique treatment modalities. Research into further applications continues to flourish ensuring that lasers retain their important role in medicine.


Amplification of emerging viruses in a bat colony.
Bats host noteworthy viral pathogens, including coronaviruses, astroviruses, and adenoviruses. Knowledge on the ecology of reservoir-borne viruses is critical for preventive approaches against zoonotic epidemics. We studied a maternity colony of Myotis myotis bats in the attic of a private house in a suburban neighborhood in Rhineland-Palatinate, Germany, during 2008, 2009, and 2010. One coronavirus, 6 astroviruses, and 1 novel adenovirus were identified and monitored quantitatively. Strong and specific amplification of RNA viruses, but not of DNA viruses, occurred during colony formation and after parturition. The breeding success of the colony was significantly better in 2010 than in 2008, in spite of stronger amplification of coronaviruses and astroviruses in 2010, suggesting that these viruses had little pathogenic influence on bats. However, the general correlation of virus and bat population dynamics suggests that bats control infections similar to other mammals and that they may well experience epidemics of viruses under certain circumstances.


Rhinocerebral zygomycosis: an unusual dramatic presentation in a paediatric cardiac patient without risk factors.
Mucormycosis is an angioinvasive infection caused by ubiquitous filamentous fungi of the order Mucorales. We describe a case of rhinocerebral mucormycosis presented in a paediatric patient after cardiac surgery correction of a complete atrioventricular canal defect. The rhinocerebral form of mucormycosis in our patient presented as an important epistaxis that needed immediate intubation due to blood inhalation. Furthermore, due to the worsening of pulmonary function, the patient also needed mechanical support with veno-arterial extracorporeal membrane oxygenation. The patient died as a consequence of a disseminated form of fungal infection. We describe our experience of this rare opportunistic infection and we think that early recognition of the disease could help in proper management.


NF-κB, JNK, and TLR Signaling Pathways in Hepatocarcinogenesis.
Hepatocellular carcinoma (HCC) is the third largest cause of cancer deaths worldwide. The role of molecular changes in HCC have been used to identify prognostic markers and chemopreventive or therapeutic targets. It seems that toll-like receptors (TLRs) as well as the nuclear factor (NF)-κB, and JNK pathways are critical regulators for the production of the cytokines associated with tumor promotion. The cross-talk between an inflammatory cell and a neoplastic cell, which is instigated by the activation of NF-κB and JNKs, is critical for tumor organization. JNKs also regulate cell proliferation and act as oncogenes, making them the main tumor-promoting protein kinases. TLRs play roles in cytokine and hepatomitogen expression mainly in myeloid cells and may promote liver tumorigenesis. A better understanding of these signaling pathways in the liver will help us understand the mechanism of hepatocarcinogenesis and provide a new therapeutic target for HCC.


Use of the focused assessment with sonography for trauma (FAST) examination and its impact on abdominal computed tomography use in hemodynamically stable children with blunt torso trauma.
The aim of this study was to evaluate the variability of clinician-performed Focused Assessment with Sonography for Trauma (FAST) examinations and its impact on abdominal computed tomography (AbCT) use in hemodynamically stable children with blunt torso trauma (BTT). The FAST is used with variable frequency in children with BTT. We performed a planned secondary analysis of children (<18 years) with BTT. Patients with a Glasgow Coma Scale (GCS) score of less than 9, those with hypotension, and those taken directly to the operating suite were excluded. Clinicians documented their suspicion for intra-abdominal injury (IAI) as very low, less than 1%; low, 1% to 5%; moderate, 6% to 10%; high, 11% to 50%; or very high, greater than 50%. We determined the relative risk (RR) for AbCT use based on undergoing a FAST examination in each of these clinical suspicion strata. Of 6,468 (median age, 11.8 years; interquartile range, 6.3-15.5 years) children who met eligibility, 887 (13.7%) underwent FAST examination before CT scan. A total of 3,015 (46.6%) underwent AbCT scanning, and 373 (5.8%) were diagnosed with IAI. Use of the FAST increased as clinician suspicion for IAI increased, 11.0% with less than 1% suspicion for IAI, 13.5% with 1% to 5% suspicion, 20.5% with 6% to 10% suspicion, 23.2% with 11% to 50% suspicion, and 30.7% with greater than 50% suspicion. The patients in whom the clinicians had a suspicion of IAI of 1% to 5% or 6% to 10% were significantly less likely to undergo a CT scan if a FAST examination was performed: RR, 0.83 (0.67-1.03); RR, 0.81 (0.72-0.91); RR, 0.85 (0.78-0.94); RR, 0.99 (0.94-1.05); and RR, 0.97 (0.91-1.05) for patients with clinician suspicion of IAI of less than 1%, 1% to 5%, 6% to 10%, 11% to 50%, and greater than 50%, respectively. The FAST examination is used in a relatively small percentage of children with BTT. Use increases as clinician suspicion for IAI increases. Patients with a low or moderate clinician suspicion of IAI are less likely to undergo AbCT if they receive a FAST examination. A randomized controlled trial is required to more precisely determine the benefits and drawbacks of the FAST examination in the evaluation of children with BTT. Prognostic and epidemiologic study, II.