Teleradiology from the provider's perspective-cost analysis for a mid-size university hospital.
Real costs of teleradiology services have not been systematically calculated. Pricing policies are not evidence-based. This study aims to prove the feasibility of performing an original cost analysis for teleradiology services and show break-even points to perform cost-effective practice. Based on the teleradiology services provided by the Greifswald University Hospital in northeastern Germany, a detailed process analysis and an activity-based costing model revealed costs per service unit according to eight examination categories. The Monte Carlo method was used to simulate the cost amplitude and identify pricing thresholds. Twenty-two sub-processes and four staff categories were identified. The average working time for one unit was 55 (x-ray) to 72 min (whole-body CT). Personnel costs were dominant (up to 68 %), representing lower limit costs. The Monte Carlo method showed the cost distribution per category according to the deficiency risk. Avoiding deficient pricing by a likelihood of 90 % increased the cost of a cranial CT almost twofold as compared with the lower limit cost. Original cost analysis is possible when providing teleradiology services with complex statutory requirements in place. Methodology and results provide useful data to help enhance efficiency in hospital management as well as implement realistic reimbursement fees. • Analysis of original costs of teleradiology is possible for a providing hospital • Results discriminate pricing thresholds and lower limit costs to perform cost-effective practice • The study methods represent a managing tool to enhance efficiency in providing facilities • The data are useful to help represent telemedicine services in regular medical fee schedules.


Maintenance of long-term potentiation in rat dentate gyrus requires protein synthesis but not messenger RNA synthesis immediately post-tetanization.
The involvement of new protein and messenger ribonucleic acid synthesis in long-term potentiation was studied in the anaesthetized rat dentate gyrus using several inhibitors of protein synthesis (anisomycin, emetine, cycloheximide and puromycin) and an inhibitor of messenger ribonucleic acid synthesis (actinomycin D). When injected for 1 h just prior to tetanization, the four inhibitors of protein synthesis produced a mild reduction of long-term potentiation of the excitatory postsynaptic potential measured 10 min after tetanization. Anisomycin produced a significantly faster decay of long-term potentiation, while the other inhibitors had more moderate effects. Actinomycin D failed to affect long-term potentiation. In a second experiment, the time-dependency of the anisomycin effect was examined. Anisomycin injected immediately after tetanization promoted decay of long-term potentiation, but when injected after a 15-min delay, the drug had no effect. Inhibition of protein synthesis for 4 h prior to tetanization did not have any more effect on long-term potentiation than inhibition for 1 h. In no experiment was long-term potentiation of the population spike affected by drug manipulation. These results suggest that for long-term potentiation of the excitatory postsynaptic potential to be maintained for at least 3 h proteins must be synthesized from already existing messenger ribonucleic acid, and that this synthesis is mostly completed within 15 min after tetanization.


Severe obstetric maternal morbidity: a 15-year population-based study.
Using a provincial perinatal database for 15 years, 1988-2002. Cases were identified with one or more of the following markers of severe maternal morbidity: blood transfusion > or = 5 units, emergency hysterectomy, uterine rupture, eclampsia, intensive care (ICU) admission. There were 159,896 mothers delivered of whom 313 (2.0/1000) had 385 markers of severe morbidity (257 had one, 42 had two, 12 had three, and two had four). The following rates of morbidity were recorded: blood transfusion > or = 5 units 119 (0.74/1000); emergency hysterectomy 88 (0.55/1000); uterine rupture 49 (0.31/1000); eclampsia 46 (0.28/1000); ICU 83 (0.52/1000). There was a statistically significant association between multiparity > or = 1, and emergency hysterectomy and uterine rupture; between age > or = 35 years, and emergency hysterectomy, uterine rupture and ICU; and between caesarean delivery and blood transfusion > or = 5 units, emergency hysterectomy, uterine rupture, eclampsia and ICU. The main contributing obstetric complications were haemorrhage (64.7%) and complications of hypertensive disorders (16.8%).


Population-based study of pelvic lymph node positivity in clinically localized prostate cancer: a study comparing African Americans and whites.
To evaluate the correlation between race and lymph node metastasis for prostate cancer by analyzing which preoperative parameters may predict lymph node status in both races. We analyzed a group of patients (552 American white men [AWM] and 423 African-American men [AAM]) who underwent radical prostatectomy plus modified pelvic lymphadenectomy between January 1991 and June 1997. Patients who received neoadjuvant radiation or hormone therapy were excluded. Univariate and multivariate analyses were performed to determine the influence of race on lymph node positivity, as well as to correlate the preoperative parameters (serum prostate-specific antigen [PSA], biopsy Gleason score, and clinical stage) with lymph node metastasis for each race separately. The AAM presented with significantly higher preoperative Gleason scores and PSA levels than AWM. However, comparing lymph node status by race, the difference of positivity (41 AWM [7.4% and 22 AAM [5.2%]) was not statistically significant (P = 0.16). The percentage of positive nodes was similar in both races for each subset of PSA, Gleason score, and clinical stage. Despite the statistical significance of the three preoperative parameters in univariate analysis, in multivariate analysis only PSA and Gleason score were independent predictors of positive lymph nodes. There is no influence of race on lymph node metastasis, despite AAM presenting with higher preoperative Gleason scores and PSA levels. In multivariate analysis, preoperative Gleason score and PSA were independent factors for positive nodes regardless of race.


Immunotherapy in aggressive B-cell lymphomas.
The idea that the immune system could be co-opted to treat cancer is not new; it has existed for centuries. However, what is new is the advancement of our understanding of how the immune system is regulated and how a tumor evolves to evade an immune response. This knowledge, combined with modern technologies to manipulate the immune system, both pharmacologically and genetically, has led to the realization of immuno-oncology as a new frontier in cancer therapeutics. This review will focus on pharmacologic immunotherapies in aggressive B cell lymphomas: checkpoint inhibition and bispecific antibodies. The success of checkpoint inhibitors in this heterogenous collection of diseases has largely been limited to those that genetic aberrations involving genes for checkpoint ligands, whereas bispecific antibodies appear to be more broadly efficacious but responses are short-lived. Investigation into the tumor microenvironment for each of the aggressive B cell lymphoma histologies, and interrogation of mechanisms of resistance as well as predictors of response to these immunotherapy approaches, will undoubtedly identify rational combinations as well as new therapeutic targets such that outcomes can be improved across these diseases.


The medical dissolution of phytobezoars using cellulase.
Phytobezoars are plant fibre concretions occurring in the stomach as a complication following some types of gastric surgery. Symptoms include distension, pain and vomiting. Ulceration, gastric bleeding and even perforation can occur. If untreated, they result in a significant mortality. Dissolution of the phytobezoar can be achieved without operative intervention. Enzymes and especially cellulase help to break up the mass and it can then be aspirated or allowed to pass on. Five patients with phytobezoars were thus treated and the cellulase method was confirmed to be simple, safe and effective.


New genes as drivers of phenotypic evolution.
During the course of evolution, genomes acquire novel genetic elements as sources of functional and phenotypic diversity, including new genes that originated in recent evolution. In the past few years, substantial progress has been made in understanding the evolution and phenotypic effects of new genes. In particular, an emerging picture is that new genes, despite being present in the genomes of only a subset of species, can rapidly evolve indispensable roles in fundamental biological processes, including development, reproduction, brain function and behaviour. The molecular underpinnings of how new genes can develop these roles are starting to be characterized. These recent discoveries yield fresh insights into our broad understanding of biological diversity at refined resolution.


Issues surrounding PACS archiving to external, third-party DICOM archives.
In larger health care imaging institutions, it is becoming increasingly obvious that separate image archives for every department are not cost effective or scalable. The solution is to have each department's picture archiving communication system (PACS) have only a local cache, and archive to an enterprise archive that drives a universal clinical viewer. It sounds simple, but how many PACS can truly work with a third-party Integration of the Health Care Enterprise Compliant Image Archive? The answer is somewhat disappointing.


Evaluating the Catalytic Potential of a General RNA-Cleaving FANA Enzyme.
The discovery of synthetic genetic polymers (XNAs) with catalytic activity demonstrates that natural genetic polymers are not unique in their ability to function as enzymes. However, all known examples of in vitro selected XNA enzymes function with lower activity than their natural counterparts, suggesting that XNAs might be limited in their ability to fold into structures with high catalytic activity. To explore this problem, we evaluated the catalytic potential of FANAzyme 12-7, an RNA-cleaving catalyst composed entirely of 2'-fluoroarabino nucleic acid (FANA) that was evolved to cleave RNA at a specific phosphodiester bond located between an unpaired guanine and a paired uracil in the substrate recognition arm. Here, we show that this activity extends to chimeric DNA substrates that contain a central riboguanosine (riboG) residue at the cleavage site. Surprisingly, FANAzyme 12-7 rivals known DNAzymes that were previously evolved to cleave chimeric DNA substrates under physiological conditions. These data provide convincing evidence that FANAzyme 12-7 maintains the catalytic potential of equivalent DNAzymes, which has important implications for the evolution of XNA catalysts and their contributions to future applications in synthetic biology.


Robotic-assisted beating heart mitral valve surgery: Preliminary report.
Robotic mitral valve surgery is a challenging issue, particularly in patients who are not suitable for aortic cross-clamping. In this study, we aimed to determine the feasibility and benefits of robotic, beating heart mitral valve surgery. From February 2019 to February 2022, 17 patients underwent robotic beating heart mitral valve surgery. Fourteen of the patients had previous cardiac surgery. The mean age was 58.1 ± 10.3. Dense periaortic adhesions, heavily calcified aorta, and low ejection fraction were retained as indications for beating heart surgery. Mitral valve replacement was performed in 14 patients. Mitral ring annuloplasty was performed in two patients with low ejection fraction (EF). A severe paravalvular leak was repaired in one patient. Additional tricuspid annuloplasties were performed in three patients. Cardiopulmonary bypass time were 185.6 ± 55 min. There were no cases of conversion to sternotomy or thoracotomy. No cerebrovascular event occurred in the follow-up. One patient died as a result of secondary hepatorenal syndrome and multiorgan failure. Robotic beating heart mitral valve surgery is a feasible and effective technique with favorable early and mid-term results, especially in patients who are not suitable for aortic cross-clamping, secondary to periaortic adhesions, severe aortic calcifications, and low ejection fraction.


Clinico-histological and molecular features of hepatocellular carcinoma from nonalcoholic fatty liver disease.
Patients with nonalcoholic fatty liver disease (NAFLD) continue to increase with the epidemics of obesity, and NAFLD is estimated to become the most prevalent etiology of hepatocellular carcinoma (HCC). Recently, NAFLD-HCC has been recognized to have clinico-histologically and molecularly distinct features from those from other etiologies, including a lower incidence rate of HCC and less therapeutic efficacy to immune checkpoint inhibitors (ICIs). Consistent with the clinical observations that up to 50% of NAFLD-HCC occurs in the absence of cirrhosis, the imbalance of pro- and antitumorigenic hepatic stellate cells termed as myHSC and cyHSC can contribute to the creation of an HCC-prone hepatic environment, independent of the absolute fibrosis abundance. Immune deregulations by accumulated metabolites in NAFLD-affected livers, such as a fatty-acid-induced loss of cytotoxic CD4 T cells serving for immune surveillance and "auto-aggressive" CXCR6+ CD8 T cells, may promote hepatocarcinogenesis and diminish therapeutic response to ICIs. Steatohepatitic HCC (SH-HCC), characterized by the presence of fat accumulation in tumor cells, ballooned tumor cells, Mallory-Denk body, interstitial fibrosis, and intratumor immune cell infiltration, may represent a metabolic reprogramming for adapting to a lipid-rich tumor microenvironment by downregulating CPT2 and leveraging its intermediates as an "oncometabolite." Genome-wide analyses suggested that SH-HCC may be more responsive to ICIs given its mutual exclusiveness with β-catenin mutation/activation that promotes immune evasion. Thus, further understanding of NAFLD-specific hepatocarcinogenesis and HCC would enable us to improve the current daily practice and eventually the prognoses of patients with NAFLD.


Genome sequence of a cluster A13 mycobacteriophage detected in Mycobacterium phlei over a half century ago.
A phage infecting Mycobacterium phlei was isolated in 1958 from a soil sample in Hungary. Some physicochemical and biological properties of the virus were described in independent studies over the years. Here, we report the genome sequence of this early mycobacteriophage isolate. The Phlei phage genome measured 50,418 bp, had a GC content of 60.1 % and was predicted to encode 81 proteins and three tRNAs. Phylogeny of the tape measure protein revealed genetic relatedness to other early isolates of mycobacteriophages within subcluster A2. The genomic organization and genetic relationships to other strains showed that the Phlei phage belongs to a novel genetic cluster, designated A13.


Chemokine CXCL1 as a potential marker of disease activity in systemic lupus erythematosus.
The chemokine CXCL1, known as growth-related oncogene α (GRO-α), is a potent chemoattractant and regulator of neutrophils. The purpose of our study was to evaluate the regulatory response of CXCL1 in the serum of patients with systemic lupus erythematosus (SLE) in the active stage of disease and to assess whether it was implicated in the pathogenesis/inflammatory process in lupus. CXCL1 serum concentrations were examined in 90 SLE patients, 56 other autoimmune diseases (OADs) patients and 100 healthy controls using enzyme-linked immunosorbent methodology. SLE patients exhibited significant increases in serum CXCL1 concentrations [1492.86 (735.47-2887.34) pg/ml] compared with OADs patients [155.88 (10.77-366.78) pg/ml] and healthy controls [13.58 (8.46-37.22) pg/ml] (p < 0.001). Moreover, the level of CXCL1 decreased as the level of anti-dsDNA IgG decreased after treatment between the anti-dsDNA-positive SLE patients and the anti-dsDNA-negative SLE patients. Additionly, serum CXCL1 concentrations were related to different disease activity levels in SLE and lupus nephritis (LN) and high avidity of IgG ANAs (HA IgG ANAs) (p < 0.05). Furthermore, CXCL1 serum concentrations were significantly correlated with the SLE Disease Activity Index(SLEDAI) score, relative avidity index (RAI) of HA IgG ANAs and the levels of anti-dsDNA IgG, CRP, ESR, albumin, C3 and C4.Additionally, Statistical analysis revealed that positivity for IgG ANA (p < 0.001), the presence of HA IgG ANAs (p = 0.001) and the logarithmic level of anti-dsDNA IgG (p = 0.021) were significantly associated with the logarithmic level of CXCL1 with standard partial regression coefficients (95% CI) of 2.371 (1.734-3.009), 1.231 (0.52-1.937) and 0.409 (0.062-0.755), respectively. Finally, using cutoff points of 1182.17 pg/mL and 1500.31 pg/mL, serum CXCL1 levels had a similar sensitivity of 76% and specificity of 100% and 75% for the diagnosis of active SLE and LN, respectively. Serum CXCL13 concentrations might represent a potential marker of disease activity in systemic lupus erythematosus.


Dexmedetomidine is Associated with an Increased Incidence of Bradycardia in Patients with Trisomy 21 After Surgery for Congenital Heart Disease.
This study aimed to evaluate adverse cardiac events using dexmedetomidine in infants with trisomy 21 and those without (controls) and examined potential risk factors in infants after cardiovascular surgery. We conducted a single-center retrospective cohort study. The medical records of 124 consecutive infants who had undergone cardiovascular surgery between April 1, 2013, and October 31, 2015, were enrolled. Clinical characteristics, usage of dexmedetomidine, and perioperative medications were analyzed. Adverse cardiac events were assessed with the Naranjo score and World Health Organization-The Uppsala Monitoring Centre (WHO-UMC) criteria. In total, 124 consecutive infants (30 patients and 94 controls) met the inclusion criteria. Eight of 30 (26.7 %) patients with trisomy 21 and 12 of 94 (12.8 %) controls experienced adverse cardiac events (i.e., hypotension, transient hypertension, and bradycardia) during dexmedetomidine with median Naranjo score of 6, and causality categories of WHO-UMC criteria were "certain" or "probable." Of those, the incidence of bradycardia occurred at a higher rate in patients with trisomy 21 than in controls (P = 0.011). Multiple logistic regression analysis revealed that the presence of trisomy 21 was an independent risk factor for adverse cardiac events of dexmedetomidine after cardiovascular surgery (odds ratio 4.10, 95 % CI 1.17-11.19, P = 0.006). Dexmedetomidine is associated with an increased incidence of bradycardia in patients with trisomy 21 after surgery for congenital heart disease. Physicians using dexmedetomidine should know a great deal about the characteristics of patients with trisomy 21, and hemodynamic monitoring should be closely observed.


Videocinematography of fresh and cryopreserved embryos: a retrospective analysis of embryonic morphology and implantation.
Eleven morphologic criteria were studied from videotapes of 323 fresh and 103 thawed embryos. The proportion of thawed embryos (30/39; 77%) with more than one abnormality was (P = 0.03) higher than that of fresh embryos (5/13; 38%), despite similar implantation rates (18% and 15%, respectively). The best predictor of thawed embryo implantation was cell-cell adherence: when scored "positive," 11 of 17 women (65%) became pregnant, whereas none became pregnant (0/15; P = 0.0002) when blastomeres did not adhere. The total number of abnormalities for thawed embryos was an important prognostic. An increased percentage variation of zona pellucida thickness was the most important prognostic for fresh embryos. When the "best" embryo had a zona pellucida that varied more than 25%, 24 of 60 (40%) resulted in pregnancy; pregnancies were not induced (0/21) when the "best" embryo had less than 10% variation (P = 0.0003).


Thiol-Ene Click Synthesis of Phenylboronic Acid-Functionalized Covalent Organic Framework for Selective Catechol Removal from Aqueous Medium.
We report a thiol-ene click strategy for the preparation of a novel phenylboronic acid-functionalized covalent organic framework (COF) for selective removal of catechol in aqueous solution. Vinyl-functionalized 2,5-diallyloxyterephthalaldehyde (Da-V) was prepared as a building ligand. Da-V was then condensed with 1,3,5-tris(4-aminophenyl)benzene (Tab) to give a vinyl-functionalized COF DhaTab-V. Subsequently, 4-mercaptophenylboronic acid (4-MPBA) was covalently linked on DhaTab-V via thiol-ene click reaction to give phenylboronic acid-functionalized COF DhaTab-PBA. The adsorption isotherms, energetics and kinetics, and reusability of DhaTab-PBA for the adsorption and removal of catechol from aqueous solution were investigated in detail. This phenylboronic acid-functionalized COF is promising as sorbent for selective removal of catechol from aqueous medium with large adsorption capacity and good reusability.


Biological variability of thyroid autoantibodies (anti-TPO and anti-Tg) in clinically and biochemically stable patients with autoimmune thyroid disease.
The biological variation of anti-TPO and anti-Tg autoantibodies was studied in 17 clinically and biochemically stable female patients with autoimmune thyroid disease (AITD), at regular monthly intervals over a period of 6 consecutive months. The mean and standard deviation (SD), within-subject coefficient of variation (CV), between-subject CV, index of individuality, reliability coefficient, and critical differences were as follows: for anti-TPO 238 (197) U/ml, 9.2%, 81.4%, 0.11, 0.96, and 27.6%; and for anti-Tg 1,785 (3,170) U/ml, 6.9%, 174%, 0.04, 0.99, and 22.3%. The data indicate a low within-subject CV, and a high between-subject CV that is particularly pronounced for anti-Tg. The high individuality of both autoantibodies indicates that an isolated result compared to conventional population-based reference intervals is of very little value for diagnosis. Furthermore, the near to 1 reliability coefficient for both autoantibodies correctly classifies the patient with respect to his or her homeostatic mean antibody concentration in a 6-month period of clinical and biochemical stability of thyroid disease. Imprecision goals for anti-TPO and anti-Tg antibodies are attainable with current methodology.


Exploring the molecular mechanisms of the involvement of GZMB-Caspase-3-GSDME pathway in the progression of rheumatoid arthritis.
Rheumatoid arthritis (RA) is an autoimmune disease with an unclear pathogenesis. Granzyme B (GZMB) has been reported as a potential therapeutic target for RA treatment, but its mechanism remains unclear. This study aimed to explore the molecular mechanism of the GZMB-Caspase-3-GSDME pathway in the progression of RA. An SD rat model of RA was constructed, and Western blot analysis was used to verify the high expression of the GZMB gene in RA rats. Functional validation was then performed on two common RA cells, HFLS-RA cells and MH7A cells, by inhibiting the GZMB gene with the GZMB siRNA virus. Cell proliferation function was measured by CCK8 and EDU assays; cell pyroptosis markers were detected by the LDH assay; inflammation factor levels were measured by ELISA; and the expression of GZMB and pathway-related genes and proteins was measured by Western blot. After GZMB silencing, cell proliferation was decreased compared to the control group, and the inflammation factors IL-1b and IL-18, as well as the pyroptosis markers LDH, IL-1b, and IL-18, were all reduced. The GZMB-related proteins GZMB, caspase-3, and Gasdermin E (GSDME) were also decreased. Therefore, GZMB silencing reduces pyroptosis by inhibiting caspase-3 and Gasdermin E decomposition. In summary, GZMB silencing inhibits the activation of caspase-3 and Gasdermin E, thereby delaying inflammation in RA. The GZMB gene may be a potential therapeutic target for RA.


T and B leukemic cell lines exhibit different requirements for cell death: correlation between caspase activation, DFF40/DFF45 expression, DNA fragmentation and apoptosis in T cell lines but not in Burkitt's lymphoma.
The execution phase of apoptosis occurs through the activation and function of caspases which cleave key substrates that orchestrate the death process. Here, we have compared the sensitivity of various T and B cell lines to death receptor or staurosporine-induced apoptosis. First, we found a lack of correlation between death receptor expression and sensitivity to Fas or Trail. By contrast, a correlation between caspase activation, DNA fragmentation and cell death in T cell lines was evidenced. Among T cells, CEM underwent apoptosis in response to CH11 but were resistant to Trail in agreement with the absence of Trail receptors (DR4 and DR5) on their surface. The B cell line SKW 6.4 was sensitive to CH11 and staurosporine but resistant to Trail. As B cell lines expressed significant levels of DR4 and DR5, resistance to Trail in SKW 6.4 is likely due to the expression of the decoy receptor DcR1. Burkitt's lymphoma such as RPMI 8866 and Raji did not exhibit DNA fragmentation in response to CH11, Trail or staurosporine but showed long-term caspase-dependent loss of viability upon effector treatment. The B cell lines used in this study express very weak or undetectable levels of DFF40 and relatively high levels of DFF45. Interestingly, cytosolic extracts from RPMI 88.66 but not other B lymphoma exhibit altered levels of cytochrome c-dependent caspase activation. Taken together, our results show that: (1) death receptor expression does not correlate with sensitivity to apoptosis; (2) the very low ratio of DFF40 vs. DFF45 is unlikely to explain by itself the lack of DNA fragmentation observed in certain B cell lines; and (3) a defective cytochrome c-dependent caspase activation might account at least in part for the insensitivity of certain Burkitt's lymphoma (RPMI 88.66) to apoptosis. Thus it seems that resistance of Burkitt's lymphoma to apoptosis is not governed by a general mechanism, but is rather multifactorial and differs from one cell line to another.


Nonresonant, femtosecond laser vaporization and electrospray post-ionization mass spectrometry as a tool for biological tissue imaging.
An ambient mass spectrometry imaging (MSI) source is demonstrated with both high spatial and mass resolution that enables measurement of the compositional heterogeneity within a biological tissue sample. The source is based on nonresonant, femtosecond laser electrospray mass spectrometry (LEMS) coupled to a quadrupole time-of-flight (QTOF) mass analyzer. No matrix deposition and minimal sample preparation is necessary for the source. The laser, translation stage, and mass spectrometer are synchronized and controlled using a customized user interface. Single or multiple laser shots may be applied to each pixel. A scanning rate of 2.0s per pixel is achieved. Measurement of a patterned ink film indicates the potential of LEMS for ambient imaging with a lateral resolution of ∼60μm. Metabolites including sugar, anthocyanins and other small metabolites were successfully mapped from plant samples without oversampling using a spot size of 60×70μm(2). Molecular identification of the detected analytes from the tissue was enabled by accurate mass measurement in conjunction with tandem mass spectrometry. Statistical analysis, non-negative matrix factorization and principle component analysis, were applied to the imaging data to extract regions with distinct and/or correlated spectral profiles.


Nuclear patterns of human breast cancer cells during apoptosis: characterisation by fractal dimension and co-occurrence matrix statistics.
An analytical strategy combining fractal geometry and grey-level co-occurrence matrix (GLCM) statistics was devised to investigate ultrastructural changes in oestrogen-insensitive SK-BR3 human breast cancer cells undergoing apoptosis in vitro. Apoptosis was induced by 1 microM calcimycin (A23187 Ca(2+) ionophore) and assessed by measuring conventional cellular parameters during the culture period. SK-BR3 cells entered the early stage of apoptosis within 24 h of treatment with calcimycin, which induced detectable changes in nuclear components, as documented by increased values of most GLCM parameters and by the general reduction of the fractal dimensions. In these affected cells, morphonuclear traits were accompanied by the reduction of distinct gangliosides and loss of unidentifiable glycolipid molecules at the cell surface. All these changes were shown to be involved in apoptosis before the detection of conventional markers, which were only measurable during the active phases of apoptotic cell death. In overtly apoptotic cells treated with 1 microM calcimycin for 72 h, most nuclear components underwent dramatic ultrastructural changes, including marginalisation and condensation of chromatin, as reflected in a significant reduction of their fractal dimensions. Hence, both fractal and GLCM analyses confirm that the morphological reorganisation of nuclei, attributable to a loss of structural complexity, occurs early in apoptosis.


[Determination of citrinin in cereals].
A simultaneous reversed-phase HPLC determination of citrinin and ochratoxin A in cereals is proposed. Both mycotoxins are eluted on a RP-amid C16 column using a gradient eluent acidified with phosphoric acid. The limits of detection, for a signal-to-background ratio of 3, are 1 µg/kg for citrinin and 0,4 µg/kg for ochratoxin A.


Electrical and chemical stimulations of the pontine micturition center.
In an acute decerebrate cat, electrical stimulation of the pontine micturition center (PMC) resulted in micturition. Carbachol injection into the PMC also resulted in micturition. The pattern of changes in bladder pressure and the sphincter activity observed during carbachol-induced micturition was almost identical to that observed during reflex micturition. The injection site corresponded to the nucleus locus coeruleus alpha (LC alpha). These results suggest that activation of cholinoceptive neurons in the LC alpha presumably becomes a trigger to recruit any one of a number of neuronal circuits involved in micturition.


Synthesis, conformational analysis and biological studies of cyclic cationic antimicrobial peptides containing sugar amino acids.
Sugar amino acid based 24-membered macrocyclic C2-symmetric cationic peptides were designed and synthesized. The cationic group was introduced in the sugar amino acids. The conformation of these cyclic compounds was ascertained through NMR techniques, which proved they were amphipathic in nature. All the compounds were bacteriolytic, showed good activity against the Gr(+ve) and Gr(-ve) bacteria, and exhibited low hemolytic activity.


Facilitating intergenerational learning between older people and student nurses: An integrative review.
To examine the literature on intergenerational learning between older people and student nurses. Intergenerational activities offer opportunities for intergenerational learning and help reduce ageism. There are several older person/school children intergenerational learning initiatives. However, there is less known about how intergenerational learning occurs in nurse education programmes outside of service provision. Whittemore and Knafl's (2005) integrative review framework was used to guide the review process. Population, intervention, context and outcome (PICO) was used to develop the review question, search strategy and inclusion/exclusion criteria. Database (CINAHL, Cochrane library, Medline, PubMed, Scopus and PsychInfo) searches and hand searching occurred from 2012 to 2023. Screening, appraisal and data extraction was undertaken according to Prisma guidelines. Nine papers were included (North American (n = 5), Canadian (n = 1) Chinese (n = 2), Taiwanese (n = 1)). Mixed methods designs were included. Four themes were identified: 1) Seeing beyond first glance; 2) Connecting and getting to know each other; 3) Learning together; and 4) Challenges for intergenerational learning. This review demonstrates the relevance of intergenerational learning in nurse education and highlights the importance of embedding initiatives which will promote and support mutual learning. Innovative intergenerational initiatives enable students to explore their underlying attitudes and views in a way that they may not be able to in the more traditional service and care giving learning situations.


The interaction between ApoA2 -265T>C polymorphism and dietary fatty acids intake on oxidative stress in patients with type 2 diabetes mellitus.
Apolipoprotein A2 (APOA2) -265T>C polymorphism has been studied in relation to oxidative stress and various dietary fatty acids. Since the interaction between APOA2 polymorphism and dietary fatty acids on oxidative stress has not yet discussed, we aimed to investigate the interaction on oxidative stress in type 2 diabetes mellitus (T2DM) patients. The subjects were 180 T2DM patients with known APOA2 genotype, either TT, TC or CC. Superoxide dismutase (SOD) activity was determined by colorimetric method. Total antioxidant capacity (TAC) and serum level of 8-isoprostane F2α were measured by spectrophotometry and ELISA, respectively. Dietary intake was collected through a food frequency questionnaire. Based on the median intake, fatty acids intake was dichotomized into high or low groups. The interaction between APOA2 polymorphism and dietary fatty acids intake was analyzed by ANCOVA multivariate interaction model. Higher than median intake of omega-6 polyunsaturated fatty acids (n-6 PUFA) was associated with increased serum level of 8-isoprostane F2α in subjects with TT/TC genotype (p = 0.004), and higher than median intake of omega-3 polyunsaturated fatty acids (n-3 PUFA) was associated with increased serum SOD activity in CC genotype (p < 0.001). There was a statistically significant interaction between APOA2 polymorphism and n-6 PUFA intake on 8-isoprostane F2α concentration as well as n-3 PUFA intake on serum SOD activity (p-interaction = 0.04 and 0.02, respectively). The current study shows the interaction between APOA2 polymorphism and dietary fatty acids intake on oxidative stress. More investigations on different populations are required to confirm the interaction.


The cell-surface proteome of cultured adipose stromal cells.
In this technical note we describe a method to evaluate the cell surface proteome of human primary cell cultures and cell lines. The method utilizes the BD Biosciences lyoplate, a system covering 242 surface proteins, glycoproteins, and glycosphingolipids plus relevant isotype controls, automated plate-based flow cytometry, conventional file-level analysis and unsupervised K-means clustering of markers on the basis of percent of positive events and mean fluorescence intensity of positive and total clean events. As an example, we determined the cell surface proteome of cultured adipose stromal cells (ASC) derived from 5 independent clinical isolates. Between-sample agreement of very strongly expressed (n = 32) and strongly expressed (n =16) markers was excellent, constituting a reliable profile for ASC identification and determination of functional properties. Known mesenchymal markers (CD29, CD44, CD73, CD90, CD105) were among the identified strongly expressed determinants. Among other strongly expressed markers are several that are potentially immunomodulatory including three proteins that protect from complement mediated effects (CD46, CD55, and CD59), two that regulate apoptosis (CD77 and CD95) and several with ectoenzymatic (CD10, CD26, CD13, CD73, and CD143) or receptor tyrosine kinase (CD140b (PDGFR), CD340 (Her-2), EGFR) activity, suggesting mechanisms for the anti-inflammatory and tissue remodeling properties of ASC. Because variables are standardized for K-means clustering, results generated using this methodology should be comparable between instrumentation platforms. It is widely generalizable to human primary explant cultures and cells lines and will prove useful to determine how cell passage, culture interventions, and gene expression and silencing affect the cell-surface proteome.


Noise-boosted bidirectional backpropagation and adversarial learning.
Bidirectional backpropagation trains a neural network with backpropagation in both the backward and forward directions using the same synaptic weights. Special injected noise can then improve the algorithm's training time and accuracy because backpropagation has a likelihood structure. Training in each direction is a form of generalized expectation-maximization because backpropagation itself is a form of generalized expectation-maximization. This requires backpropagation invariance in each direction: The gradient log-likelihood in each direction must give back the original update equations of the backpropagation algorithm. The special noise makes the current training signal more probable as bidirectional backpropagation climbs the nearest hill of joint probability or log-likelihood. The noise for injection differs for classification and regression even in the same network because of the constraint of backpropagation invariance. The backward pass in a bidirectionally trained classifier estimates the centroid of the input pattern class. So the feedback signal that arrives back at the input layer of a classifier tends to estimate the local pattern-class centroid. Simulations show that noise speeded convergence and improved the accuracy of bidirectional backpropagation on both the MNIST test set of hand-written digits and the CIFAR-10 test set of images. The noise boost further applies to regular and Wasserstein bidirectionally trained adversarial networks. Bidirectionality also greatly reduced the problem of mode collapse in regular adversarial networks.


Patterns of nasalance in a sample of normal gerontologic subjects.
The purpose of this investigation was to evaluate velopharyngeal adequacy in gerontologic subjects using an acoustic analysis strategy. Sixty normal older subjects (30 male, 30 female), ranging in age from 50 to 80, read three passages developed by Fletcher (1976) that differ in number of nasal sounds. In addition, all subjects sustained the vowel /a/. The speech samples were analyzed using TONAR II, which calculates the ratio of nasal sound pressure level to nasal + oral sound pressure level (nasalance). Tonagrams (plots of nasalance over time) were obtained from all subjects. Results demonstrated that older subjects generally evidenced abnormally high nasalance values for the Zoo Passage (contains no nasal sounds). In addition, females exhibited significantly higher nasalance values than males. Inspection of the tonagrams revealed three abnormal acoustic patterns: (1) pervasively high nasalance, (2) momentary loss of velar control, (3) progressive deterioration in velar control. These results are discussed with reference to known neuroanatomical changes associated with advancing age.


Structural insights into species-specific features of the ribosome from the pathogen Staphylococcus aureus.
The emergence of bacterial multidrug resistance to antibiotics threatens to cause regression to the preantibiotic era. Here we present the crystal structure of the large ribosomal subunit from Staphylococcus aureus, a versatile Gram-positive aggressive pathogen, and its complexes with the known antibiotics linezolid and telithromycin, as well as with a new, highly potent pleuromutilin derivative, BC-3205. These crystal structures shed light on specific structural motifs of the S. aureus ribosome and the binding modes of the aforementioned antibiotics. Moreover, by analyzing the ribosome structure and comparing it with those of nonpathogenic bacterial models, we identified some unique internal and peripheral structural motifs that may be potential candidates for improving known antibiotics and for use in the design of selective antibiotic drugs against S. aureus.


Conjunctival inclusion cyst following sub-Tenon's local anaesthetic injection.
Sub-Tenon's local anaesthesia has become a popular technique of infiltrative anaesthesia for small incision cataract surgery. We report the development of an inclusion cyst of the conjunctiva following this technique. The aetiology of this complication and how it can be avoided are discussed.


A mathematically related singularity and the maximum size of protein domains.
In a paper titled "A topologically related singularity suggests a maximum preferred size for protein domains" (Zbilut et al., Proteins 2007;66:621-629), Zbilut et al. claim to have found a singularity in certain geometrical properties of protein structures, and suggest that this singularity may limit the maximum size of protein domains. They find further support for the singularity in their analysis of G-factors calculated by the PROCHECK program. Here, we show that the claimed singularity is a mathematical artifact with no physical meaning, and we reanalyze the G-factors to show that Zbilut et al.'s results are due to a single outlier in the data. Thus, the existence of an actual singularity in the topological properties of proteins is not supported by the findings of Zbilut et al.


Elevation of GM2 ganglioside during ethanol-induced apoptotic neurodegeneration in the developing mouse brain.
GM2 ganglioside in the brain increased during ethanol-induced acute apoptotic neurodegeneration in 7-day-old mice. A small but a significant increase observed 2 h after ethanol exposure was followed by a marked increase around 24 h. Subcellular fractionation of the brain 24 h after ethanol treatment indicated that GM2 increased in synaptic and non-synaptic mitochondrial fractions as well as in a lysosome-enriched fraction characteristic to the ethanol-exposed brain. Immunohistochemical staining of GM2 in the ethanol-treated brain showed strong punctate staining mainly in activated microglia, in which it partially overlapped with staining for LAMP1, a late endosomal/lysosomal marker. Also, there was weaker neuronal staining, which partially co-localized with complex IV, a mitochondrial marker, and was augmented in cleaved caspase 3-positive neurons. In contrast, the control brain showed only faint and diffuse GM2 staining in neurons. Incubation of isolated brain mitochondria with GM2 in vitro induced cytochrome c release in a manner similar to that of GD3 ganglioside. Because ethanol is known to trigger mitochondria-mediated apoptosis with cytochrome c release and caspase 3 activation in the 7-day-old mouse brain, the GM2 elevation in mitochondria may be relevant to neuroapoptosis. Subsequently, activated microglia accumulated GM2, indicating a close relationship between GM2 and ethanol-induced neurodegeneration.


The methodological quality was low and conclusions discordant for meta-analyses comparing proximal humerus fracture treatments: a meta-epidemiological study.
To investigate the association between methodological quality and reported conclusions of meta-analyses comparing operative with non-operative treatments for proximal humerus fractures. Cross-sectional meta-epidemiological study. We searched EMBASE, PubMed, The Cochrane Library, and Web of Science for systematic reviews with meta-analyses comparing non-operative with operative treatments for proximal humerus fractures. Methodological quality was assessed using AMSTAR2 and the reported conclusions were scored for three outcome domains (functional outcome, quality of life, and harm) on a scale from 1 to 6. The Mann-Whitney and Kruskal-Wallis tests were used to investigate the association between methodological quality and reported conclusions. We included 21 systematic reviews: 19 pairwise meta-analyses and 2 network meta-analyses, although there are only 8 published randomized controlled trials. Most (n = 18) of the meta-analyses were rated as critically low quality, while the remaining 1 was rated as high quality. The conclusions were discordant for all three outcome domains, even for meta-analyses reporting similar inclusion criteria. We could not perform most of the statistical tests due to the predominantly critically low quality. The methodological quality was so predominantly critically low that it was not possible to evaluate the association between methodological quality and reported conclusions.


The Biology of Huntingtin.
Huntingtin (HTT) is now a famous protein because an abnormal expansion of a glutamine stretch (polyQ) in its N-terminal sequence leads to the devastating neurodegenerative disorder Huntington's disease (HD). The gene encoding huntingtin, HTT, and its dominantly inherited mutation were identified more than 20 years ago. Subsequently, in the hope of finding a cure for HD, there has been intense research aimed at understanding the molecular mechanisms underlying the deleterious effects of the presence of the abnormal polyQ expansion in HTT. Notwithstanding with the value of this approach, evidence has been emerging of a potential role of context and function of the HTT protein in the specificity and severity of the pathogenicity. HTT is ubiquitous both at the tissue and subcellular levels. It interacts with many partners and has long been considered having no clearly defined cellular function. Based on research over the past 20 years, specifically focused on the function of wild-type HTT, we reconsider the literature describing HTT-regulated molecular and cellular mechanisms that could be dysfunctional in HD and their possible physiological consequences for patients.


Neopterin and kynurenine-tryptophan ratio as predictors of coronary events in older adults, the Hordaland Health Study.
Immune system activation is involved in atherosclerosis. Neopterin production and tryptophan catabolism through the kynurenine pathway, measured by the kynurenine-tryptophan ratio (KTR), are induced by interferon gamma, thus both are considered markers of cell mediated immune activation. This study prospectively investigated their predictive value on acute coronary events among Norwegian community-dwelling older adults without previous coronary heart disease. 1112 men and 1631 women, 71-74 years old were examined during 1997-99 as part of the Hordaland Health Study. They were followed until an acute coronary event (defined as unstable angina, non-fatal or fatal acute myocardial infarction or sudden death) or December 31, 2006. Kaplan-Meier hazard curves were constructed for quartiles of plasma neopterin and KTR. Cox proportional hazards models adjusted for sex, body mass index, smoking, hypertension, renal function and cholesterol were used to examine the relation between neopterin and KTR quartiles and the study endpoint. Median (interquartile range) values were 8.6 (7.2-10.4) nmol/L for neopterin and 25.8 (25.3-31.1) nmol/μmol for KTR. During the follow up, 265 participants had at least one acute coronary event. Increased baseline levels of plasma neopterin and KTR were associated with continuous increased risk of developing the study endpoint (P-values for trend <0.001 and 0.019, respectively). Adjusted hazard ratios comparing the fourth quartile to the first were 1.65 (95% CI; 1.11-2.47; P=0.013) for neopterin and 1.57 (95% CI 1.03-2.39; P=0.036) for KTR. Plasma neopterin and KTR levels predict acute coronary events in older adults without previous coronary heart disease.


A conserved central region of yeast Ada2 regulates the histone acetyltransferase activity of Gcn5 and interacts with phospholipids.
The SAGA (Spt-Ada-Gcn5 acetyltransferase) complex of Saccharomyces cerevisiae contains more than 20 components that acetylate and deubiquitylate nucleosomal histones. Its acetyltransferase, Gcn5, preferentially acetylates histones H3 and H2B and is regulated through interactions with Ada2 and Ngg1/Ada3. Sequence alignments of Ada2 homologs indicate a conserved approximately 120-amino-acid-residue central region. To examine the function of this region, we constructed ada2 alleles with mutations of clustered conserved residues. One of these alleles, ada2-RLR (R211S, L212A, and R215A), resulted in an approximately threefold reduction in transcriptional activation of the PHO5 gene and growth changes that parallel deletion of ada2. Microarray analyses further revealed that ada2-RLR alters expression of a subset of those genes affected by deletion of ada2. Indicative of Ada2-RLR affecting Gcn5 function, Ada2-RLR resulted in a decrease in Gcn5-mediated histone acetylation in vitro to a level approximately 40% that with wild-type Ada2. In addition, in vivo acetylation of K16 of histone H2B was almost totally eliminated at Ada2-regulated promoters in the ada2-RLR strain, while acetylation of K9 and K18 of histone H3 was reduced to approximately 40% of wild-type levels. We also show that the central region of Ada2 interacts with phospholipids. Since phosphatidylserine binding paralleled Ada2 function, we suggest that lipid binding may play a role in the function or regulation of the SAGA complex.


Effects of HCN2 Mutations on Dendritic Excitability and Synaptic Plasticity: A Computational Study.
Several reports of augmented hyperpolarisation-activated cyclic nucleotide-gated (HCN) currents in seizures have suggested a pro-convulsive identity for HCN channels. The mutations identified in one or more of the four HCN channel subunits are found to be contributing to different epileptic phenotypes. S126L, S632W, V246M and E515K are four different mutations affecting the HCN2 subunit and have been reported in febrile seizures and partial/generalised idiopathic epilepsies. From the visible outcomes in subjects with these mutations, it is evident that they must play important roles in altering dendritic excitability. Through this simulation study using NEURON, we created a three-compartmental, hippocampal CA1 pyramidal neuron synapse model expressing seven different ion channels (fast sodium (NaF), T-type calcium (CaT), R-type calcium (CaR), delayed rectifier potassium (KDR), A-type potassium (KA), small conductance potassium (SK), and HCN channels) and two glutamate receptors (AMPAR and NMDAR). We modelled an HCN2 channel and incorporated changes in it to obtain mutation kinetics. Their effects on excitability were studied by observing resting membrane potentials, input resistances and plasticity profiles for measuring the sliding modification threshold (SMT) of Bienenstock-Cooper-Munro (BCM) theory. Virtual knockouts of ion channels other than HCN were also performed to assess their role in altering excitability when they act alongside HCN2 mutations. Our results show that HCN2 mutations can potentially be a primary causative factor for excessive action potential firing through their effect on resting membrane potentials and input resistance.


Effective committees.
A committee is a group of people officially delegated to perform a function, such as investigating, considering, reporting, or acting on a matter. Some committees function like task forces and work on specific, time-limited projects. When the work is finished, the committees are no longer needed. These committees are called ad hoc committees. Other committees are standing committees. They are created by the standing orders, rules, by-laws, or regulations of an organization and exist and function indefinitely (eg, finance, membership, education, nomination). Both types of committees can form subcommittees if the workloads are heavy or complex in nature. Committees can be among the most important working forces of an organization. They serve as work units of the organization, taking work and breaking it into meaningful and manageable chunks. They efficiently carry out the work of the organization. Committee work should be a rewarding experience for both the members and the organization. Committees represent, involve, and serve members, as well as provide an important training ground for future leaders of an organization. New or inexperienced members can gain valuable insight into an organization and develop confidence by serving on committees. There are several key elements of effective committees, including (1) a clear, written purpose; (2) an effective committee chair; (3) thoughtfully appointed members; and (4) well-run meetings.


Phase I trial of high-dose infusional hydroxyurea, high-dose infusional 5-fluorouracil and recombinant interferon-alpha-2a in patients with advanced malignancies.
The ribonucleotide reductase inhibitor, hydroxyurea (HU), augments the cytotoxic effects of 5-fluorouracil (5FU) in vitro; both drugs are synergistic with interferon-alpha (IFN) in vitro. The aim of this phase I study was to determine the maximal duration of HU, 4.3 g/m2, administered as a parenteral infusion in combination with 5FU, 2.6 g/m2 administered over 24 hrs each week, + IFN, 9 MU, subcutaneously three times per week. There were 26 patients enrolled and evaluable. This included 14 patients with colorectal cancer of whom 13 had been previously treated, and 12 patients with other refractory malignancies (pancreas, cholangiocarcinoma, hepatocellular carcinoma, renal cell carcinoma, and others), of whom 10 were previously untreated. The dose-limiting toxicity of this regimen was myelosuppression. This prohibited dose escalation of HU above the starting dose (24 hrs) on a 6-weeks-on, 2-weeks-off therapy schedule. When filgrastim, 480 microg, was administered subcutaneously on days 3-6, the duration of HU could be extended to 48 hrs on a 2-weeks-on, 1-week-off therapy schedule. There were two instances of fatal infection, one in a patient with a rectovaginal fistula with neutropenic sepsis and the second in a patient with non-neutropenic Clostridium septicum sepsis. All therapy was administered in the ambulatory setting. There were three responders, all among previously untreated patients. High-dose parenteral hydroxyurea, 4.3 g/m2 administered over 24 hrs, can be safely combined with high-dose weekly 5FU, 2.6 g/m2 over 24 hrs + IFN, 9 MU subcutaneously three times per week, without filgrastim in the ambulatory setting. Parenteral hydroxyurea, 4.3 g/m2 over 24 hrs daily x 2 can also be combined with high-dose 5FU + IFN, but requires the addition of filgrastim to avoid severe myelosuppression.


Selection and characterization of spinetoram resistance in field collected Drosophila melanogaster.
Insecticides are commonly employed in vineyards to control vinegar flies and limit sour rot disease. Widespread resistance to available insecticides is having a negative impact on managing Drosophila melanogaster populations, rendering control of sour rot more difficult. An insecticide registered for use in vineyards to which resistance is not yet widespread (at least in New York and Missouri) is spinetoram. Spinetoram targets the nicotinic acetylcholine receptor α6, and mutations in α6 have been associated with resistance in some insects. Our goals were to select for a spinetoram resistant strain of D. melanogaster (starting with field collected populations), characterize the resistance, and identify the mutation responsible. After five selections a strain (SpinR) with >190-fold resistance was obtained. Resistance could not be overcome by insecticide synergists, suggesting an altered target site was involved. We cloned and sequenced the α6 allele from the spinetoram resistant strain and identified a mutation causing a glycine to alanine change at amino acid 301 (equivalent position to the G275E mutation found in some spinosad/spinetoram resistant insects). This mutation was found at low levels in field populations, but increased with each selection until it became homozygous in SpinR. We discuss how the identification of the spinetoram resistance mutation can be used for resistance management.


Fractionation and characterization of DNA at sites of replication from rat liver nuclei.
Regenerating rat liver nuclei when sonicated and centrifuged in a Cs2SO4 gradient were fractionated into three distinct bands. These bands were designated as light band (LB), middle band (MB), and heavy band (HB) according to their density. LB and MB were shown to consist of large granular particles with varying electron densities, but LB also contained remnants of nuclear membrane. When analysed by gel electrophoresis, LB and MB displayed more than 35 bands of proteins. The third fraction, HB, consisted largely of small chromatin fibers and its proteins were predominantly the four histones of the nucleosomal core particle. Following short pulses with [3H]thymidine in vivo, the specific activity of DNA in LB and MB was significantly higher than that of bulk DNA contained in HB. DNA in all three fractions became equally labelled as the duration of the labelling interval increased beyond 30 min. Newly synthesized DNA was characterized by electrophoresis on analytical 1.7% acrylamide -0.5% agarose composite gels. After a 1-min labelling interval in vivo, 17% of the rapidly labelled DNA from LB and MB was stationary at the gel origin like replication forks from E. coli, but only 3% of HB DNA had zero mobility. Electron microscopy confirmed the presence of DNA replication forks in LB, MB, and HB. With increasing time of synthesis the proportion of labelled DNA exhibiting zero mobility decreased in all three fractions. Denaturation of DNA or digestion of single-stranded DNA with S1 nuclease released newly synthesized DNA from the gel origin. Ribonuclease was without effect. DNA recovered from LB and MB also had a higher molecular weight than the HB DNA. Together these results indicate (1) that LB and MB are enriched in newly replicated DNA; (2) that an increased proportion of newly replicated DNA in LB and MB is associated with DNA replication forks; and (3) that the replicating DNA recovered in LB and MB may be associated with other nuclear constituents in situ because this DNA appears to be protected from the more frequent chain breaks introduced into the bulk of chromatin (HB) by sonication.


Human high-affinity Fc gamma RI (CD64) gene mapped to chromosome 1q21.2-q21.3 by fluorescence in situ hybridization.
The human Fc gamma RI gene encodes for a high-affinity Fc gamma receptor that plays pivotal roles in the immune response. We have used fluorescence in situ hybridization analysis to localize the Fc gamma RI gene to human chromosome 1. The human Fc gamma RI (CD64) gene has been assigned to human chromosome 1q21.2-q21.3 using R-banded human (pro)metaphase chromosomes.


Extended-depth spectral-domain optical coherence tomography imaging of the crystalline lens in Weill-Marchesani-like syndrome.
A 26-year-old woman presented with signs and symptoms of acute angle-closure glaucoma (ACG) in her left eye. Extended-depth spectral-domain optical coherence tomography (SD-OCT) was performed to biometrically assess the anterior chamber depth and crystalline lens conformation. Extended-depth SD-OCT imaging of the crystalline lens showed bilateral spherophakia without dislocation. Neither eye manifested a highly myopic refractive error despite spherophakia. Unanticipated conformation of the anterior and posterior lenticular curvatures, increased thickness of the crystalline lenses, and short axial lengths explained the lack of a highly myopic refractive error. These findings demonstrated a lens-related cause of the ACG. The use of extended-depth SD-OCT supported the diagnosis of subclinical Weill-Marchesani syndrome in a patient without other commonly associated findings and provided a better understanding of the mechanism of angle closure in this patient.


Glaucoma management among individuals enrolled in a single comprehensive insurance plan.
To determine the management patterns for glaucoma and suspect glaucoma in a nationally representative sample of newly treated persons. Retrospective cohort study of persons enrolled in a large managed care organization. One thousand seven hundred twelve diagnosed suspects and 3623 diagnosed glaucoma patients. Linked pharmacy and patient care data were used to examine the glaucoma management and treatment patterns in this cohort of persons insured by a single managed care organization. Rates of monitoring and treatment were calculated for the 3 study groups. Probability of monitoring (return visits, visual fields [VFs], and optic nerve head imaging or photography) and treatment (argon laser trabeculoplasty [ALT] and surgery) for newly treated persons with suspect and glaucoma diagnoses. After a median follow-up of 440 days, 83% of treated diagnosed suspects had had a billed follow-up office visit to either an optometrist or an ophthalmologist at any time during follow-up, 46% had had at least one billed VF, and 13% had had some form of optic nerve head imaging. Rates were slightly higher for those with diagnosed glaucoma (P>0.05). Surgery and ALT were performed rarely in this treated population (1%-6% at 2 years). This study suggests that a large proportion of individuals felt to require treatment for glaucoma or suspect glaucoma are falling out of care and are being monitored at rates lower than expected from recommendations of published guidelines. More research is needed to confirm these findings and to determine the reasons for loss to follow-up and low monitoring rates.


MuRF1-dependent regulation of systemic carbohydrate metabolism as revealed from transgenic mouse studies.
Under various pathophysiological muscle-wasting conditions, such as diabetes and starvation, a family of ubiquitin ligases, including muscle-specific RING-finger protein 1 (MuRF1), are induced to target muscle proteins for degradation via ubiquitination. We have generated transgenic mouse lines over-expressing MuRF1 in a skeletal muscle-specific fashion (MuRF1-TG mice) in an attempt to identify the in vivo targets of MuRF1. MuRF1-TG lines were viable, had normal fertility and normal muscle weights at eight weeks of age. Comparison of quadriceps from MuRF1-TG and wild type mice did not reveal elevated multi-ubiquitination of myosin as observed in human patients with muscle wasting. Instead, MuRF1-TG mice expressed lower levels of pyruvate dehydrogenase (PDH), a mitochondrial key enzyme in charge of glycolysis, and of its regulator PDK2. Furthermore, yeast two-hybrid interaction studies demonstrated the interaction of MuRF1 with PDH, PDK2, PDK4, PKM2 (all participating in glycolysis) and with phosphorylase beta (PYGM) and glycogenin (both regulating glycogen metabolism). Consistent with the idea that MuRF1 may regulate carbohydrate metabolism, MuRF1-TG mice had twofold elevated insulin blood levels and lower hepatic glycogen contents. To further examine MuRF1's role for systemic carbohydrate regulation, we performed glucose tolerance tests (GTT) in wild type and MuRF1-TG mice. During GTT, MuRF1-TG mice developed striking hyperinsulinaemia and hepatic glycogen stores, that were depleted at basal levels, became rapidly replenished. Taken together, our data demonstrate that MuRF1 expression in skeletal muscle re-directs glycogen synthesis to the liver and stimulates pancreatic insulin secretion, thereby providing a regulatory feedback loop that connects skeletal muscle metabolism with the liver and the pancreas during metabolic stress.


Health and environmental safety of nanomaterials: O Data, Where Art Thou?
Nanotechnology keeps drawing attention due to the great tunable properties of nanomaterials in comparison to their bulk conventional materials. The growth of nanotechnology in combination with the digitization era has led to an increased need of safety related data. In addition to safety, new data-driven paradigms on safe and sustainable by design materials are stressing the necessity of data even more. Data is a fundamental asset to the scientific community in studying and analysing the entire life-cycle of nanomaterials. Unfortunately, data exist in a scattered fashion, in different sources and formats. To our knowledge, there is no study focusing on aspects of actual data-structure knowledge that exists in literature and databases. The purpose of this review research is to transparently and comprehensively, display to the nanoscience community the datasets readily available for machine learning purposes making it convenient and more efficient for the next users such as modellers or data curators to retrieve information. We systematically recorded the features and descriptors available in the datasets and provide synopsised information on their ranges, forms and metrics in the supplementary material.


Pooled evidence from preclinical and clinical studies for stem cell-based therapy in ARDS and COVID-19.
COVID-19 has severely devastated many lives across the globe. It has been speculated that stem cell-based therapy for COVID-19 treatment could be able to subsidize the effects. In preclinical and clinical studies, stem cell-based therapy has successfully eliminated inflammatory cytokines in ALI, ARDS, and COVID-19. Clinical trials have produced a variety of promising results for validating stem cell therapy in COVID-19 patients. For instance, exosome-based therapy (ExoFlow) showed an 87% survival status, and MSC-based therapy (Mesoblast) achieved an 83% survival rate in moderate to severe COVID-19 patients. This review debates the advantages of cell-free therapy, i.e., stem cell-derived exosome-based therapies, over stem cell-based therapy. This review aims to question whether the immunomodulatory effect of stem cells differs based on their origin and also tries to find possible answers for the best stem cells for treating SARS-CoV-2 infection. The role of stem cells and their extracellular vesicles in the upregulation of regulatory immune cells, growth factors (EGF, FGF, VEGF), and anti-inflammatory cytokines (IL-6, INF-α, galectin-1, notch-1, PDL-1) that promote the tissue regeneration at the injured site. The right side of the image depicts the downregulation of inflammation-inducing immune cells, pro-inflammatory cytokines, and chemokines that could also enhance COVID-19 therapy.


Hepato-erythropoietic porphyria presenting as scleroderma and acrosclerosis in a sibling pair.
Clinical and biochemical data are reported on a sibling pair with clinical and biochemical features of both erythropoietic protoporphyria and hepatic cutaneous porphyria. After many years of photosensitivity the clinical picture resembles systemic sclerosis.


Rupture of the right ventricular free wall. An unusual complication of mediastinitis after cardiac surgery.
Of 2171 patients who underwent open heart surgery between 1981 and 1986, 41 (1.8%) developed postoperative mediastinitis and were treated by closed irrigation drainage or open chest therapy. Six patients, (mean age 59 years) developed rupture of the right ventricular free wall. The primary procedures were: resection of akinetic fibrous plaque (2), mitral valve replacement (1), coronary bypass grafting (1), removal of a left atrial myxoma (1) and repair of post-infarction rupture of the left ventricle (1). In 2 patients, rupture occurred in the operating theatre during revision of the irrigation drainage. Both patients died after repair. In 4 patients, rupture occurred during coughing. One died before surgery. In the 3 other cases, the defects were repaired either by direct suture (2 patients) or with a pericardial patch (1 patient) with the aid of normothermic extracorporeal circulation. Three days later, a muscular flap (pectoral or dorso-lumbar) was mobilized to protect the mediastinal viscera. All 3 patients are alive and well. When bleeding occurs during treatment of mediastinitis, an immediate exploration with extracorporeal circulation to close the defect should be considered.


The effect of methyltransferase inhibitors on histamine release from human dispersed lung mast cells activated with anti-human IgE and calcium ionophore A23187.
Inhibitors of adenosylmethionine (AdoMet)-dependent methyltransferases reduce histamine release from enzymatically dispersed human lung mast cells activated with either anti-human IgE or calcium ionophore A23187. The IC25 values for adenosine and 3-deazaadenosine (DZA) inhibiting anti-IgE-induced histamine release were 395 microM and 301 microM respectively. The addition of homocysteine thiolactone (Hcy) potentiated the effects of adenosine and DZA, reducing their IC25 values to 32 microM and 10.5 microM respectively. The adenosine deaminase (adenosine aminohydrolase EC 188.8.131.52) inhibitors erythro-9-(2-hydroxy-3-nonyl)-adenine (EHNA) inhibited anti-IgE-induced histamine release with an IC50 of 162 microM. This inhibition was not potentiated by Hcy. The combination of DZA and Hcy effectively inhibited histamine release induced by concentrations of A23187 which released a similar amount of histamine to anti-IgE. However the combination was 17 times less potent against A23187-compared with anti-IgE-induced release. These observations suggest that AdoMet-dependent methyltransferases play an important role in IgE-dependent histamine release from human lung mast cells but their role in A23187-induced release is less clear.


Urea-Derivative Catalyzed Enantioselective Hydroxyalkylation of Hydroxyindoles with Isatins.
The enantioselective transformations of indoles preferentially take place in the more-reactive azole ring. However, the methods for the enantioselective functionalization of the indole benzene ring are scarce. In this paper, a series of bifunctional (thio)urea derivatives were used to organocatalyze the enantioselective Friedel-Crafts hydroxyalkylation of indoles with isatins. The resulting products were obtained in good yields (65-90%) with up to 94% enantiomer excess (ee). The catalyst type and the substrate scope were broadened in this methodology.


Photoinhibition and recovery in tropical plant species: response to disturbance.
Disturbance or rainforest is often followed by mass mortality of understorey seedlings. Transitions of shade grown plants to full sunlight can cause reductions in the efficiency with which light is used in photosynthesis, called photoinhibition. In order to assess the influence of photoinhibition on mortality and growth after rainforest disturbance this study examined photoinhibition in both simulated and real forest disturbances in northern Papua New Guinea. In an experiment simulating rainforest disturbance, exposure of shade-grown plants to full sunlight resulted in abrupt decreases in the chlorophyll fluorescence parameter F v/F m that is characteristic of photoinhibition. However, in the well-watered plants used in these experiments there were no fatalities during 3 weeks after exposure to full sunlight. Thus, it is unlikely that photoinhibition, alone, is responsible for seedling fatalities after rainforest disturbances, but more likely that fatalities are due to photoinhibition in conjunction with other environmental stress. There were differences between the response of species to the simulated disturbance that concurred with their preferred habitats. For example, species form the genus Barringtonia, which is commonly found in shaded understorey environments, underwent greater reductions in F v/F m and were slower to recover than species that usually inhabit high solar radiation environments. The extent of photoinhibition and the rate of recovery appeared to be dependent on avoidance of direct solar radiation by altering leaf angles and on increasing maximum photosynthetic rates. A field survey of photoinhibition in man-made rainforest gaps corroborated the findings of the simulated disturbance experiment showing that plant species commonly found in shaded environments showed a greater degree of photoinhibition in forest gaps at midday than those species which are classified as species that benefit from gaps or specialist gap inhabitors.


Precise measurement for line structure light vision sensor with large range.
High precision and large measurement range are the target of any one three-dimensional scanner. For a line structure light vision sensor, measurement precision depends on its calibration results, i.e., determining mathematical expression of the light plane in camera coordinate system. However, as calibration results are locally optimal solutions, high precise measurement in a large range is difficult. In this paper, we give a precise measurement method and the corresponding calibration procedure for a line structure light vision sensor with a large measurement range. A motorized linear translation stages with a travel range of 150 mm and a planar target which is a surface plate with a machining precision of 0.05 mm are utilized. With the help of the linear translation stage and the planar target, functions which gives the relationship between center point of the laser stripe and the perpendicular/ horizontal distance are obtained. Once image of light stripe is captured, we can get a precise measurement result from the normalized feature points. Compared with a traditional measurement method, distortion compensation is not necessary and precision of measurement is improved significantly. Experiments show that root mean square error of measurement results according to our proposed method is reduced by 64.67% related to the traditional method.


An in vitro DNA phosphorothioate modification reaction.
Phosphorothioation (PT) involves the replacement of a nonbridging phosphate oxygen on the DNA backbone with sulfur. In bacteria, the procedure is both sequence- and stereo-specific. We reconstituted the PT reaction using purified DndCDE from Salmonella enterica and IscS from Escherichia coli. We determined that the in vitro process of PT was oxygen sensitive. Only one strand on a double-stranded (ds) DNA substrate was modified in the reaction. The modification was dominant between G and A in the GAAC/GTTC conserved sequence. The modification between G and T required the presence of PT between G and A on the opposite strand. Cysteine, S-adenosyl methionine (SAM) and the formation of an iron-sulfur cluster in DndCDE (DndCDE-FeS) were essential for the process. Results from SAM cleavage reactions support the supposition that PT is a radical SAM reaction. Adenosine triphosphate (ATP) promoted the reaction but was not essential. The data and conclusions presented suggest that the PT reaction in bacteria involves three steps. The first step is the binding of DndCDE-FeS to DNA and searching for the modification sequence, possibly with the help of ATP. Cysteine locks DndCDE-FeS to the modification site with an appropriate protein conformation. SAM triggers the radical SAM reaction to complete the oxygen-sulfur swapping.


[Epidemiology and diagnosis of urinary lithiasis: a cross-sectional study in a Cameroonian based population].
urinary stone is a multifactorial disease characterized by the presence of concretions in the urinary tract. The diagnosis of urinary lithiasis is based on clinical and imaging findings. The objective of this study was to determine the profile of patients suffering from urolithiasis in the city of Yaounde. this was a descriptive observational cross-sectional study with retrospective data collection. Patients with an age > 15 years and patients diagnosed with lithiasis confirmed by medical imaging were included in our study. The variables studied were sociodemographic, clinical and paraclinical. a total of 120 patients were included in our study. The mean age was 40.46±12.62 years with extremes from 19 to 74 years. Male gender was predominant at 60.8% (n=73). Renal colic was the circumstance found in 67.5% (n=81). Physical examination was normal in 55.8% (n=67). Insufficient hydration was found in 45% (n=61) of cases. Abdominal scanner was the most performed examination in our study in 50.7% (n=61). The median stone density was 731 Hounsfield units [346; 1183.5]. The stones were predominantly located in the upper tract. The left renal topography was predominant in 35% (n=42) of cases. Urine culture revealed Escherichia Coli at 60% (n=15). Blood calcium, phosphorus and uricemia were requested in 15.8% (n=19), 0.8% (n=1) and 12.5% (n=15) of cases, respectively. The results were normal. In the absence of a laboratory specialized in biochemical analysis of calculi, a minority of patients (n=3) benefited from spectrophotometry. urinary lithiasis is a disease of men in their forties. Renal colic is the main revealing sign. The diagnosis is revealed by the abdominal scanner in half of the cases. Metabolic assessment and constitutional analysis of the calculus are still very rarely requested.


Double recording system of Period1 gene expression rhythm in the olfactory bulb and liver in freely moving mouse.
Clock genes express circadian rhythms in most organs. These rhythms are organized throughout the whole body, regulated by the suprachiasmatic nucleus (SCN) in the brain. Disturbance of these clock gene expression rhythms is a risk factor for diseases such as obesity and cancer. To understand the mechanism of regulating clock gene expression rhythms in vivo, multiple real time recording systems are required. In the present study, we developed a double recording system of Period1 expression rhythm in peripheral tissue (liver) and the brain. In peripheral tissue, quantification of gene expression in a steadily moving target was achieved by using a photomultiplier tube (PMT) attached to a tissue contact optical sensor (TCS). Using this technique, we were able to analyze circadian rhythms of clock gene expression over a prolonged period in the liver and olfactory bub (OB) of the brain. The present double recording system has no effect on behavioral activity or rhythm. Our novel system thus successfully quantifies clock gene expression in deep areas of the body in freely moving mice for a period sufficient to analyze circadian dynamics. In addition, our double recording system can be widely applied to many areas of biomedical research, as well as applications beyond medicine.


Detection and clinical outcome of small-for-gestational-age fetuses in the third trimester-A comparison between routine ultrasound examination and examination on indication.
Fetal growth restriction is a major risk factor for adverse perinatal outcome. As most of the growth-restricted fetuses are small for gestational age (SGA), an efficient antenatal screening method for SGA fetuses would have a major impact on perinatal health. The aim of this study was to compare the SGA prediction rate achieved with third-trimester routine ultrasound estimation of fetal weight (EFW) with that obtained using ultrasound examination on indication. The secondary aim was to evaluate the clinical outcome in relation to the SGA screening method. During 1995-2009, two perinatal centers in southern Sweden offered routine ultrasound examination at 32-34 gestational weeks to 99 265 women with singleton pregnancies. Of these, 59 452 (60%) underwent the ultrasound examination. The other population, comprising 24 868 pregnancies, was cared for in another three centers that used a risk-based method with ultrasound examinations on indication only. Of them, 5792 (23%) underwent ultrasound examination at 32-36 gestational weeks. The deviation in the EFW from the expected one was expressed as the EFW z-score, SGA EFW being defined as the EFW z-score less than -2. SGA prediction ability was assessed by receiver operating characteristic (ROC) curves. Crude and adjusted risk ratios were calculated for selected variables of perinatal outcome when comparing the populations. The SGA prediction ability for routine ultrasound was high, area under the ROC curve was 0.90 (95% CI 0.89-0.91). For an EFW z-score of -1, the sensitivity was 67.3% and specificity was 90.5% among routinely screened pregnancies; corresponding numbers in the ultrasound on indication population were 34.3% and 96.6%. The screened population had a lower risk of preterm birth, birthweight z-score less than -3, and Apgar score less than 7 at 5 min with adjusted risk ratios 0.87 (95% CI 0.82-0.92), 0.75 (95% CI 0.61-0.92), and 0.77 (95% CI 0.68-0.87), respectively. No difference in perinatal mortality was detected. There were no differences in perinatal outcome between the two subcohorts of infants born SGA. Third-trimester routine ultrasound improves the detection of SGA antenatally compared with ultrasound performed on indication, but no convincing improvement in perinatal outcome was identified.


Regulatory mechanisms of IL-2 and IFNgamma suppression by quercetin in T helper cells.
Quercetin is a popular flavonoid compound that is biosynthesized by plants; it is suggested to modulate a variety of inflammatory responses of macrophages and T lymphocytes. Oral administration of quercetin in arthritic rats dramatically diminishes clinical signs of arthritis. Moreover, quercetin ameliorates experimental autoimmune encephalomyelitis, which is associated with Th1-mediated immune responses. Like quercetin inhibits macrophage-induced cytokine production, it also blocks IL-12-dependent JAK-STAT signaling in Th cells. Despite the anti-inflammatory effects of quercetin acting through Th cells, the regulatory mechanisms remain unclear. Here, we studied the function of quercetin in Th cells and found that quercetin suppressed both IFNgamma and IL-2 production upon T cell receptor stimulation. Furthermore, we uncovered the regulatory mechanisms of quercetin involved in the inhibition of cytokine production during Th cell activation. The fact that quercetin-derived IFNgamma suppression was blocked in T-bet-deficient Th cells demonstrated quercetin act through the modulation of T-bet expression. Whereas IL-2 inhibition by quercetin was independent of T-bet expression, quercetin diminished IL-2R alpha expression, which is critical for positive regulatory loop of IL-2 autoactivation. Taken together, quercetin is suggested to repress both IFNgamma and IL-2 cytokine production by independent mechanisms; T-bet-dependent IFNgamma suppression and IL-2R alpha-dependent IL-2 inhibition.


Plasma interleukin-6 levels correlate with survival in patients with bacterial sepsis and septic shock.
In this study, IL-6 levels were assessed as inflammatory biomarker of bacterial sepsis in patients hospitalized at the ICU of the hospital of Colombia. Prospective study on 62 patients diagnosed with sepsis and septic shock. An ELISA assay was used to test serum levels of IL-6 at admission and 48 h after admission. Variables were analyzed by χ2 test (alfa <0.05). Multivariable Cox regression was used to determine the survival with the statistical program SPSS v23.00. Patient's median age was 53 years old and 59.7% were male. Lung was the most common primary site of infection (43.5%), and hypertension comorbidity with higher prevalence (40%). Infection by Gram negative bacteria were significantly more frequent among patients than Gram positive (P = 0.037). Overall, survival analysis showed that 10 (16.1%) patients died with a survival median of 7.00 +4.874 (2-3) days. In patients with sepsis we detected a significant decline in the average of IL-6 serum levels after 48 h of admission [7.50 (SD: 7.00-68.00) pg/mL vs. 68.00 [SD: 7.00-300.00] pg/mL (P = 0.000). Only 25% of patients with septic shock who presented high levels of IL-6 at the time of admission and at 48 h had a survival up to 15 days (P = 0.005). We found significant differences between the plasma levels of IL-6 during the first 48 h after admission to the ICU among patients with sepsis and septic shock. Patients with sepsis had a significant decline in IL-6 levels, whereas in patients who developed septic shock, levels of this cytokine remained high and have a lower survival compared to those who maintained low levels of IL-6.


Entrepreneurial patent management in pharmaceutical startups.
Startups fill an increasingly important role as innovators in the pharmaceutical industry, and patenting is typically central to their success. This article aims to explore patent management in pharmaceutical startups. The results show that startups need to deal with several challenges related to patenting and an 'entrepreneurial' approach to patent management is called for. Resource constraints, venture capital provision, exits and other conditions and events must be readily considered in the patent management process to build a successful pharmaceutical venture, something that could benefit the pharmaceutical industry as a whole.


Downregulation of PHEX in multibacillary leprosy patients: observational cross-sectional study.
Peripheral nerve injury and bone lesions, well known leprosy complications, lead to deformities and incapacities. The phosphate-regulating gene with homologies to endopeptidase on the X chromosome (PHEX) encodes a homonymous protein (PHEX) implicated in bone metabolism. PHEX/PHEX alterations may result in bone and cartilage lesions. PHEX expression is downregulated by intracellular Mycobacterium leprae (M. leprae) in cultures of human Schwann cells and osteoblasts. M. leprae in vivo effect on PHEX/PHEX is not known. Cross-sectional observational study of 36 leprosy patients (22 lepromatous and 14 borderline-tuberculoid) and 20 healthy volunteers (HV). The following tests were performed: PHEX flow cytometric analysis on blood mononuclear cells, cytokine production in culture supernatant, 25-hydroxyvitamin D (OHvitD) serum levels and (99m)Tc-MDP three-phase bone scintigraphy, radiography of upper and lower extremities and blood and urine biochemistry. Significantly lower PHEX expression levels were observed in lepromatous patients than in the other groups (χ(2) = 16.554, p < 0.001 for lymphocytes and χ(2) = 13.933, p = 0.001 for monocytes). Low levels of 25-(OHvitD) were observed in HV (median = 23.0 ng/mL) and BT patients (median = 27.5 ng/mL) and normal serum levels were found in LL patients (median = 38.6 ng/mL). Inflammatory cytokines, such as TNF, a PHEX transcription repressor, were lower after stimulation with M. leprae in peripheral blood mononuclear cells from lepromatous in comparison to BT patients and HV (χ(2) = 10.820, p < 0.001). Downregulation of PHEX may constitute an important early component of bone loss and joint damage in leprosy. The present results suggest a direct effect produced by M. leprae on the osteoarticular system that may use this mechanism.


Alternative promoter usage and mRNA splicing pathways for parathyroid hormone-related protein in normal tissues and tumours.
The parathyroid hormone-related protein (PTHrP) gene consists of nine exons and allows the production of multiple PTHrP mRNA species via the use of three promoters and 5' and 3' alternative splicing; as a result of 3' alternative splicing one of three protein isoforms may be produced. This organisation has potential for tissue-specific splicing patterns. We examined PTHrP mRNA expression and splicing patterns in a series of tumours and normal tissues, using the sensitive reverse transcription-polymerase chain reaction (RT-PCR) technique. Use of promoter 3 and mRNA specifying the 141 amino acid PTHrP isoform were detected in all samples. Transcripts encoding the 139 amino acid isoform were detected in all but two samples. Use of promoters 1 and 2 was less widespread as was detection of mRNA encoding the 173 amino acid isoform. While different PTHrP splicing patterns were observed between tumours, no tissue- or tumour-specific transcripts were detected. In comparing normal and tumour tissue from the same patient, an increase in the number of promoters utilised was observed in the tumour tissue. Furthermore, mRNA for the PTH/PTHrP receptor was detected in all samples, thus the PTHrP produced by these tumours may potentially act in an autocrine or paracrine fashion.


Peripheral blood lymphocytes with receptors for a determinant common to B-lymphoblastoid cell lines and acute myelogenous leukemia blasts.
We investigated whether the rosetting of B-lymphoblastoid cell lines (B-LCL) by peripheral blood lymphocytes (PBLs) reflected possible interactions between lymphoid cells and immature cells of the hematopoietic system. Rosette formation could be blocked by the addition of soluble antigen extracted from B-LCL or blasts obtained from patients with acute myelogenous leukemia (AML). This inhibition was specific for AML blasts (similarly extracted material from melanoma lines had no inhibitory effect) and for the B-LCL receptor (leukemic extracts had no effect on surface receptors for sheep red blood cells (E) or antibody-sensitized red blood cells (EA)). The B-LCL receptor is present on leukemic Sezary T-cells as well as normal T-cells and its sensitivity to various enzymatic treatments is markedly different from that of E and EA receptors. In addition, B-LCLs derived from in vitro EB-viral infection of a normal donor's B lymphocytes were significantly rosetted by that donor's autologous PBLs. These data suggests the B-LCL receptor, present on mature T-cells, can recognize self determinants on myeloblasts and B-LCL. Further investigation will determine whether this interaction can affect the function of rosetted target cells.


T cell receptor-V beta repertoire in allergen-specific sensitization and increased airway responsiveness.
An animal model system was developed to study the mechanisms resulting in allergic sensitization. Local allergen exposure via the airways and the lung stimulated an allergen-specific immunoglobulin E (IgE) response that was paralleled by the development of increased airway responsiveness (AR). It was found that CD4+ T cells of local draining lymph nodes played an important role in the regulation of these events. Stimulation of allergen-specific T cells requires interaction between major histocompatibility complex (MHC) class II molecules (expressed on antigen-presenting cells), peptide (presented on MHC) and the T cell receptor. Allergen sensitization stimulated T cells that expressed a restricted T cell receptor V beta (TCR-V beta) elements. Each allergen stimulated different V beta elements, and sensitization to the same allergen resulted in a different pattern of TCR-V beta stimulation in different lymphoid tissues. Some of these T cells had pro-allergenic effects, whereas others were able to inhibit the development of the allergic response, including the development of increased AR. These data indicate that the local T cell response regulates the type of immune response that evolves following local allergen sensitization.


Effect of intravenous epinephrine infusion on plasma renin activity in adrenalectomized dogs.
Previous experiments have shown that circulating epinephrine stimulates renin secretin and increases plasma renin activity (PRA) when it is infused intravenously, but not when it is infused directly into the renal artery at similar infusion rates. The present experiments were designed to test the hypothesis that the adrenal glands mediate the PRA response to intravenous epinephrine infusion. Accordingly, anesthetized dogs were prepared with either an acute bilateral adrenalectomy or a sham-adrenalectomy procedure. Epinephrine was then infused intravenously into each animal for 45 minutes at a rate of 25 ng X kg-1 X min-1. Time control experiments in which epinephrine was not infused were also conducted. In sham-adrenalectomized dogs, PRA (in nanograms per ml h-1) rose from 4.1 +/- 1.4 in the control period to 13.0 +/- 3.0 during intravenous epinephrine infusion (means +/- SE; p less than 0.01). In adrenalectomized dogs, PRA rose from 2.1 +/- 0.4 during the control period to 5.5 +/- 0.9 during intravenous epinephrine infusion (p less than 0.01). Neither the absolute increments in PRA nor the percent increases in PRA were significantly different between the two groups receiving epinephrine. PRA remained unchanged in time control experiments. These data demonstrate that the adrenal glands need not be present in order for intravenous epinephrine infusion to elicit an increase in PRA. The data do not support the hypothesis, therefore, that epinephrine-induced increases in PRA are initiated by receptors located within the adrenal glands.


Fast determination of prominent carotenoids in tomato fruits by CEC using methacrylate ester-based monolithic columns.
In this study, the major carotenoids (beta-carotene and lycopene) present in tomato fruits were analyzed by CEC with a methacrylate ester-based monolithic column. The effects of the porogenic solvent ratio, and the hydrophobicity of bulk monomer employed were examined on carotenoids separations. A fast separation of these analytes was achieved in less than 5.0 min in a mobile phase containing 35% THF, 30% ACN, 30% methanol, and 5% of a 5 mM Tris aqueous buffer, pH 8, with lauryl methacrylate-based monoliths. The CEC method was evaluated in terms of detection limit and reproducibility (retention time, area, and column preparation) with values below 1.6 microg/mL and 7.2%, respectively. The proposed procedure was successfully applied to the determination of both carotenoids in fruits of several tomato-related species and its usefulness to analyze large series of samples for nutritional quality screening trials in tomato breeding programs is demonstrated. To our knowledge, this is the first work that exploits the powerful and user-friendly monolithic technology for quality breeding and germplasm evaluation program purposes.


Algorithm in choosing recipient vessels for perforator free flap in breast reconstruction: the role of the internal mammary perforators.
Although the internal mammary (IM) vessels are our first choice as recipient vessels for free flap breast reconstruction, attempts to reduce surgical morbidity led us to adopt an algorithm when choosing recipient vessels. The IM vessels give direct perforators to the breast. They can be found either superficial or deep to the pectoralis major muscle. If they are of a reasonable calibre they can be considered as recipient vessels. When the perforators are not suitable, the IM vessels are used for the microanastomosis. This protocol was adopted for our patients between June 1999 and December 2002. Breast reconstruction with free flaps was performed on 298 patients with perforator flaps. IM and thoracodorsal (TD) vessels are used in 88% and 3% of cases, respectively. IM perforators were successfully used in 30 cases (9%). The IM perforators used were located at the level of the second and third intercostal space in nine (30%) and 21 (70%) cases, respectively. The average diameters of those perforators were 1 mm (0.5-1.3 mm) for the artery and 1.7 mm (1-3 mm) for the vein. The IM perforators can be used as recipient vessels for free flaps. This spares the IM vessels for eventual cardiac bypass surgery, avoiding recipient site morbidity and decreasing the postoperative discomfort.


Receptor-independent, vacuolar ATPase-mediated cellular uptake of histamine receptor-1 ligands: possible origin of pharmacological distortions and side effects.
The aims of this study were to investigate whether several histamine receptor agonists and antagonists are subjected to receptor-independent ion trapping into acidic organelles, and whether this sequestration influences their pharmacological or toxicological properties. Vacuolar (V)-ATPase-dependent intracellular sequestration of agonists was recognized as morphological alterations (large fluid-filled vacuoles for betahistine and 1-methylhistamine, granular uptake for fluorescent BODIPY FL histamine) prevented by the specific V-ATPase inhibitor bafilomycin A1 in rabbit vascular smooth muscle cells. Lipophilicity was the major determinant of these cellular effects (order of potency: BODIPY FL histamine>betahistine>1-methylhistamine>histamine) that occurred at high concentrations. This ranking was dissociable from the potency order for H(1) receptor-mediated contraction of the rabbit aorta, a response uninfluenced by bafilomycin. Antihistamines are inherently more lipophilic and caused vacuolization of a proportion of cells at 5-500 microM. Agonist or antagonist-induced vacuoles were of macroautophagic nature (labeled with GFP-conjugated LC3, Rab7 and CD63; detection of LC3 II). Further, the 2 most lipophilic antihistamines tested, astemizole and terfenadine, were potentiated by V-ATPase blockade in the aortic contractility assay (13- and 3.6-fold more potent, respectively, pA(2) scale), suggesting that V-ATPase-mediated cation trapping sequesters these antagonists from the vicinity of H(1) receptors in the therapeutic concentration range. This potentiation did not apply to less lipophilic antagonists (pyrilamine, diphenhydramine). While some agonists and all tested antagonists of the histamine H(1) receptors induce the V-ATPase-dependent vacuolar and autophagic cytopathology, sequestration affects the pharmacology of only the most lipophilic antagonists, the ones prone to off-target arrhythmogenic side effects.


Outcome of glomerulonephritis in live-donor renal transplant recipients: A single-centre experience.
To investigate the frequency and risk factors affecting the incidence of post-transplantation glomerulonephritis (GN) and the impact of GN on the survival of the graft and the patient. Patients were classified based on histological findings into three groups. Graft survival was ascertained using the Kaplan-Meier method and significance calculated using log-rank tests. For multivariate analysis the Cox model was used. Transplant glomerulopathy was the most prevalent glomerular disease in our series followed by recurrent GN and lastly de novo GN. In all, 50% of the de novo GN group had diabetes. The worst graft outcomes were in the recurrent GN group (P = 0.044). Multivariate analysis revealed ageing of the graft and mammalian target of rapamycin (mTOR) immunosuppression as risk factors for development of GN. While, the age of the recipient and donor, anti-lymphocyte globulin induction therapy, and acute rejection were risk factors for poor graft outcomes. GN is an important issue after transplantation. Tracking the incidence and progression of histological findings in the graft may help to guide proper management and improve graft outcome.


Assessment of Marine Mammal Impact Zones for Use of Military Sonar in the Baltic Sea.
Military sonars are known to have caused cetaceans to strand. Navies in shallow seas use different frequencies and sonar pulses, commonly frequencies between 25 and 100 kHz, compared with most studied NATO sonar systems that have been evaluated for their environmental impact. These frequencies match the frequencies of best hearing in the harbor porpoises and seals resident in the Baltic Sea. This study uses published temporary and permanent threshold shifts, measured behavioral response thresholds, technical specifications of a sonar system, and environmental parameters affecting sound propagation common for the Baltic Sea to estimate the impact zones for harbor porpoises and seals.