Surgical approach after failed enteryx injection for GERD.
We report on 3 patients who underwent laparoscopic antireflux procedures for persistent symptoms of GERD after biopolymer injection. Experienced laparoscopic surgeons completed all 3 procedures laparoscopically. In 2 patients, there was an extramural extravasation of the polymer outside and adherent to the esophageal wall. In these patients, a partial posterior fundoplication was used. The third patient, who had the polymer material deposits removed preoperatively by endoscopic mucosal resection, underwent a Nissen fundoplication. Postoperative recovery was uneventful in all cases. At follow-up of 6 to 12 months, all patients were symptom free, off medical therapy, and experiencing no dysphagia. Surgical therapy for patients after failed biopolymer injection is safe and effective. The choice of surgery may depend on whether the polymer mass can be removed preoperatively.


Structural and thermodynamic studies on cation-Pi interactions in lectin-ligand complexes: high-affinity galectin-3 inhibitors through fine-tuning of an arginine-arene interaction.
The high-resolution X-ray crystal structures of the carbohydrate recognition domain of human galectin-3 were solved in complex with N-acetyllactosamine (LacNAc) and the high-affinity inhibitor, methyl 2-acetamido-2-deoxy-4-O-(3-deoxy-3-[4-methoxy-2,3,5,6-tetrafluorobenzamido]-beta-D-galactopyranose)-beta-D-glucopyranoside, to gain insight into the basis for the affinity-enhancing effect of the 4-methoxy-2,3,5,6-tetrafluorobenzamido moiety. The structures show that the side chain of Arg144 stacks against the aromatic moiety of the inhibitor, an interaction made possible by a reorientation of the side chain relative to that seen in the LacNAc complex. Based on these structures, synthesis of second generation LacNAc derivatives carrying aromatic amides at 3'-C, followed by screening with a novel fluorescence polarization assay, has led to the identification of inhibitors with further enhanced affinity for galectin-3 (K(d) > or = 320 nM). The thermodynamic parameters describing the binding of the galectin-3 C-terminal to selected inhibitors were determined by isothermal titration calorimetry and showed that the affinity enhancements were due to favorable enthalpic contributions. These enhancements could be rationalized by the combined effects of the inhibitor aromatic structure on a cation-Pi interaction and of direct interactions between the aromatic substituents and the protein. The results demonstrate that protein-ligand interactions can be significantly enhanced by the fine-tuning of arginine-arene interactions.


Antiangiogenic therapy in lung cancer: focus on vascular endothelial growth factor pathway.
Lung cancer (LC) is a leading cause of death worldwide. Recent advances in chemotherapeutic agents have not yielded any significant improvement in the prognosis of patients with LC. The five-year survival rate for all combined disease stages remains about 15%. For this reason, new therapies such as those that inhibit tumor angiogenesis or block activity of growth factor receptors are of special interest in this group of patients. In this review we will summarize the most recent clinical data on biologic therapies that inhibit tumor angiogenesis in LC, focusing on those that are most clinically relevant.


[Contribution of a functional orthopedic splint to the treatment of Class II malocclusions].
The numerous therapeutic, psychological and functional benefits that derive from using a two-stage approach to the treatment of Class II malocclusions can sometimes be partially offset by difficulties stemming from the choice of technique: patients may be reluctant to wear cumbersome appliances which, when they are worn, can interfere with speech production, the quality of therapeutic improvement may be reduced by undesirable dento-alveolar responses to orthopedic forces, treatment results may turn out to be unstable at the final finishing stage and continuation of treatment at the time of the appearance of the young adult dentition may be jeopardized. The author has designed a new appliance, a functional orthopedic splint whose slim design largely eliminates these problems. Suitable for use in a variety of techniques, this easily worn device greatly reduces unwanted dento-alveolar responses as it stimulates desirable functional improvement and gives orthodontists much improved control of their treatment of Class II malocclusions.


Evaluation of MicroScan Rapid Pos Combo panels for identification of staphylococci.
MicroScan Rapid Pos Combo panels (Baxter Diagnostics, Inc., MicroScan, West Sacramento, Calif.) contain substrates conjugated with fluorophores and substrates with a fluorescent pH indicator. AutoSCAn W/A, an automated panel processor equipped with a fluorometer, reads the panels after 2 h of incubation and can identify staphylococci to the species level. We tested 239 strains belonging to 17 species of staphylococci. All the strains were identified by conventional methods (W.E. Kloos and K.H. Schleifer, J. Clin. Microbiol. 1:82-88, 1975) and by the MicroScan Rapid ID system. The system correctly identified 219 (91.6%) strains; nine (3.8%) identification results were probably correct, and six (2.5%) results were incorrect. The system designated five (2.1%) strains as rare biotypes. The automated MicroScan Rapid ID system is useful and reliable in identifying most human isolates of staphylococci encountered in the clinical laboratory.


Synthesis of helical π-conjugated polymers bearing pyridine N-oxide pendants and asymmetric allylation of aldehydes in the helical cavity.
Catalytically active chiral π-conjugated polymers (poly-1(NO)r) bearing pyridine N-oxide pendants were synthesized by ternary copolymerization of a d-glucose-bound diethynyl compound with two types of thieno[3,4-b]thiophene comonomer, one of which contained a pyridine N-oxide group. When the pyridine N-oxide content in the copolymer was 10 mol% (poly-1(NO)0.10), the polymer backbone formed a one-handed helical structure in acetonitrile. Pyridine N-oxide pendants arranged inside the helical cavity of poly-1(NO)0.10 exhibited catalytic activity for the asymmetric allylation of benzaldehydes, producing the corresponding allyl alcohols with up to 43% ee.


The early stimulation of glycolysis by epidermal growth factor in isolated rat hepatocytes is secondary to the glycogenolytic effect.
We have studied the relationship between the effect of epidermal growth factor (EGF) on glycogen metabolism and its effect on glycolysis, in rat hepatocyte suspensions. Although 10 nM glucagon or 10 microM adrenaline increased glycogen degradation by more than 120%, 10 nM EGF increased glycogenolysis by less than 20% in hepatocytes incubated in glucose-free medium. Both glucagon and adrenaline increased phosphorylase a activity by more than 130%; EGF increased this activity by about 30%. Under basal conditions, 65% of the glucosyl residues were released as free glucose and about 30% ended up as C3 molecules (lactate and pyruvate). Both glucagon and adrenaline decreased the proportion of glucosyl units that rendered glycolysis end-products (to 2% for glucagon and 6% for adrenaline) and increased the proportion that ended up as free glucose (to 94% and 88% of the glucosyl residues for glucagon and adrenaline respectively). EGF increased the production of both free glucose and lactate+pyruvate, but the proportion of glucosyl residues that ended up as free glucose or glycolysis end-products was unchanged. In glycogen-depleted hepatocytes incubated in the presence of 25 mM glucose, EGF affected neither glycogen deposition nor glycolysis. EGF increased cytosolic free Ca2+, and neomycin decreased both the Ca2+ signal and the glycogenolytic effect. In conclusion, our results indicate that the effect of EGF on glycolysis is secondary to the Ca(2+)-mediated stimulation of glycogenolysis in rat hepatocyte suspensions.


Polyclonal antibody to soman-tyrosine.
Soman forms a stable, covalent bond with tyrosine 411 of human albumin, with tyrosines 257 and 593 in human transferrin, and with tyrosine in many other proteins. The pinacolyl group of soman is retained, suggesting that pinacolyl methylphosphonate bound to tyrosine could generate specific antibodies. Tyrosine in the pentapeptide RYGRK was covalently modified with soman simply by adding soman to the peptide. The phosphonylated-peptide was linked to keyhole limpet hemocyanin, and the conjugate was injected into rabbits. The polyclonal antiserum recognized soman-labeled human albumin, soman-mouse albumin, and soman human transferrin but not nonphosphonylated control proteins. The soman-labeled tyrosines in these proteins are surrounded by different amino acid sequences, suggesting that the polyclonal recognizes soman-tyrosine independent of the amino acid sequence. Antiserum obtained after 4 antigen injections over a period of 18 weeks was tested in a competition ELISA where it had an IC50 of 10(-11) M. The limit of detection on Western blots was 0.01 μg (15 picomoles) of soman-labeled albumin. In conclusion, a high-affinity, polyclonal antibody that specifically recognizes soman adducts on tyrosine in a variety of proteins has been produced. Such an antibody could be useful for identifying secondary targets of soman toxicity.


Immunological screening of a glycoprotein antigen expressed by Zajdela ascites hepatoma cells on normal rat tissues and tumour cells.
Expression of the glycoprotein MII2 antigen originally identified in Zajdela ascites hepatoma cells was investigated in several normal rat tissues and in more or less differentiated tumours using biochemical and immunological approaches. SDS-polyacrylamide gel electrophoresis followed by fluorography or immunoblotting with an antiserum raised against the purified MII2 antigen revealed that this antigen was absent from normal liver cells. ELISA assays, indirect immunofluorescence and immunoprecipitation experiments using the same antiserum showed that this glycoprotein was not expressed in various normal tissues such as liver, spleen, lung, pancreas, intestine and stomach, but it was unexpectedly detected in kidney and thymic tissues. However, the molecular weight of the antigens immunoprecipitated from kidney and thymus was lower than the one of MII2 (Mr of 60,000 versus 110,000-160,000 for purified MII2). No staining was observed in embryonic rat liver at 10 and 20 days of development. Moreover, this antigen was present on the surface of Morris hepatoma 7777, another rapidly proliferating and poorly differentiated hepatocellular carcinoma. In contrast, this antigen was not detected on the surface of in vitro Zajdela hepatoma cells (ZHC) or of partially differentiated hepatomas (Faza) which have recovered some hepatic functions. In addition, the MII2 antigen was found on the human non-hepatic HT-29 tumour cell line, under its undifferentiated form (HT-29 G+ subline). The possible relationships between the expression of this antigen and both the malignant transformation process and the differentiation process are discussed.


Prevention of autoimmune recurrence and rejection by adenovirus-mediated CTLA4Ig gene transfer to the pancreatic graft in BB rat.
Type 1 diabetes is the result of a selective destruction of pancreatic islets by autoreactive T-cells. Therefore, in the context of islet or pancreas transplantation, newly transplanted beta-cells are threatened by both recurrent autoimmune and alloimmune responses in recipients with type 1 diabetes. In the present study, using spontaneously diabetic BB rats, we demonstrate that whereas isolated islets are susceptible to autoimmune recurrence and rejection, pancreaticoduodenal grafts are resistant to these biological processes. This resistance is mediated by lymphohematopoietic cells transplanted with the graft, since inactivation of these passenger cells by irradiation uniformly rendered the pancreaticoduodenal grafts susceptible to recurrent autoimmunity. We further studied the impact of local immunomodulation on autoimmune recurrence and rejection by ex vivo adenovirus-mediated CTLA4Ig gene transfer to pancreaticoduodenal grafts. Syngeneic DR-BB pancreaticoduodenal grafts transduced with AdmCTLA4Ig were rescued from recurrent autoimmunity. In fully histoincompatible LEW-->BB transplants, in which rejection and recurrence should be able to act synergistically, AdmCTLA4Ig transduced LEW-pancreaticoduodenal allografts enjoyed markedly prolonged survival in diabetic BB recipients. In situ reverse transcription-polymerase chain reaction revealed that transferred CTLA4Ig gene was strongly expressed in both endocrine and exocrine tissues on day 3. These results indicate the potential utility of local CD28-B7 costimulatory blockade for prevention of alloimmune and autoimmune destruction of pancreatic grafts in type 1 diabetic hosts.


Biological clock dysfunction exacerbates contact hypersensitivity in mice.
Immediate-type skin allergic reactions, such as passive cutaneous anaphylactic reaction, are associated with circadian rhythm, but the role of circadian mechanisms on delayed-type skin allergic reactions, such as contact hypersensitivity (CHS), remains uncertain. In mice, CHS, a T-cell-mediated immune response, is a classic model of human allergic contact dermatitis. We investigated whether biological clock dysfunction affects CHS pathogenesis in CLOCK mutant mice compared with wild-type (WT) mice. Mice were treated with 2,4,6-trinitro-1-chlorobenzene (TNCB) on the abdominal skin on day 0 (sensitization) and then treated with TNCB on the ears on day 5 (challenge). We found that biological clock dysfunction resulted in severe inflammation. Ear swelling, serum immunoglobulin E level and mast cell number were significantly increased in CLOCK mutant mice compared with WT mice. These results provide evidence that CLOCK mutation promotes the T-helper type 2 immune response and exacerbates CHS. Corticosterone has a protective effect on CHS. The serum corticosterone level lost rhythmicity and showed a decreased daily level in CLOCK mutant mice compared with WT mice, supporting the exacerbating effect of CLOCK mutation on CHS. Adrenalectomy markedly worsened TNCB-induced CHS in WT mice but not in CLOCK mutant mice. In addition, dramatic dexamethasone-induced protection of CHS was observed in CLOCK mutant mice compared with WT mice. The present results suggest that circadian rhythm might be an important factor in the regulation of CHS via corticosterone rhythmicity and/or level.


Postsynaptic alpha adrenergic receptor subtypes differentiated by yohimbine in tissues from the rat. Existence of alpha-2 adrenergic receptors in rat aorta.
The selective alpha-2 adrenergic receptor antagonist, yohimbine, was used to differentiate postsynaptic alpha-adrenergic receptors in five peripheral tissues of the rat. Three distinct postsynaptic receptor subtypes were observed based on the affinity of the receptors for yohimbine. Receptors with high affinity for yohimbine were detected in the aorta, whereas low affinity receptors were observed in the vas deferens. The affinity for yohimbine in these two tissues differed by over 50-fold. Receptors with intermediate affinity for yohimbine were found in the portal vein, spleen and bladder. The Schild plot for yohimbine in the bladder suggests that blockade of alpha receptors in this tissue is not competitive or that there exists more than one type of alpha-adrenergic receptor present. Comparison of the dissociation constant of yohimbine in the aorta with dissociation constants obtained from the literature for this compound in a variety of tissues containing alpha-1 or alpha-2 adrenergic receptors indicates that the postsynaptic alpha adrenergic receptor in the aorta is of the alpha-2 type. Conversely, the postsynaptic alpha adrenergic receptor in the portal vein appears to be alpha-1. The differences in postsynaptic alpha appears to be alpha-1. The differences in postsynaptic alpha adrenergic receptors in these two vascular tissues may reflect the marked differences in adrenergic innervation and the possible relative lack of neuroeffector junctional alpha adrenergic receptors in the rat aorta.


Computed tomography of the pharynx in a closed vs. open mouth position.
The pharynx is anatomically complex and evaluation can be difficult even with cross-sectional imaging. Eight animals had computed tomography (CT) studies of the head performed with the mouth open and closed. The studies were anonymized and evaluated by four radiologists for visibility of six anatomic regions (dorsal wall of nasopharynx, lumen of nasopharynx, dorsal margin of the soft palate, ventral margin of the soft palate, oropharynx, and laryngopharynx) and for certainty of a normal or abnormal diagnosis of four different anatomic regions (nasopharynx, soft palate, oropharynx, and laryngopharynx). Mean visual scores differed significantly between mouth positions and were improved when the mouth was open. The ability of radiologists to classify anatomic regions as normal or abnormal vs. unsure also varied between mouth positions, and there was greater uncertainty when the mouth was closed. In addition, estimated volume of the air-filled nasopharynx differed significantly as a function of mouth position and was greater when the mouth was open (mean=1.187 cm(3) , SE=0.177) vs. closed (mean=0.584 cm(3) , SE=0.116). Computed tomographic evaluation of the pharynx can be improved with the mouth open.


Access to care among patients with osteogenesis imperfecta during the COVID-19 pandemic.
In a cross-sectional study assessing the experiences of individuals with osteogenesis imperfecta accessing care during the COVID-19 pandemic, participants reported high rates of delays in accessing medical care and high utilization of telehealth. Considering the needs of individuals with complex medical conditions is important when improving access to care. Individuals with osteogenesis imperfecta (OI) often have complex care needs requiring that they see a variety of specialists. The onset of the COVID-19 pandemic in March 2020 led to delays in medical care for many health conditions. The goal of this study was to describe the experiences of individuals with OI accessing medical care during this time. Responses to an electronic survey distributed via the OI Foundation mailing list were collected from August 2020 until February 2021. Participants were instructed to compare their experiences in the months since the start of the pandemic with their experiences prior to this date. Data were analyzed using descriptive statistics and were compared across demographic groups using logistic regression and chi-squared tests. Surveys were completed by 110 participants. Most participants (72%) reported experiencing delays in accessing at least one care provider. The majority of participants reported less or similar amounts of bone pain (74.3%) and less or the same rate of fracture (88.6%) as before the start of the pandemic. While most study participants experienced delays in care, they did not report an increase in symptoms associated with OI. They also frequently utilized telehealth as a tool to see their providers. Future research should focus on the impact of changes in telehealth legislation on patients' ability to access care. As methods for care delivery evolve, the needs of people with OI and other rare diseases should be considered and prioritized.


Recording information on protein complexes in an information management system.
The Protein Information Management System (PiMS) is a laboratory information management system (LIMS) designed for use with the production of proteins in a research environment. The software is distributed under the CCP4 licence, and so is available free of charge to academic laboratories. Like most LIMS, the underlying PiMS data model originally had no support for protein-protein complexes. To support the SPINE2-Complexes project the developers have extended PiMS to meet these requirements. The modifications to PiMS, described here, include data model changes, additional protocols, some user interface changes and functionality to detect when an experiment may have formed a complex. Example data are shown for the production of a crystal of a protein complex. Integration with SPINE2-Complexes Target Tracker application is also described.


The Healthy Heart-Mind Trial: Randomized Controlled Trial of Melatonin for Prevention of Delirium.
Delirium is a serious medical condition with increased incidence in at-risk surgical populations. We sought to determine if melatonin use reduces the incidence of delirium in individuals undergoing major cardiac surgery. Randomized double-blind placebo-controlled clinical trial (two arms, 1:1 allocation, parallel design). The trial took place in two metropolitan hospitals (public tertiary and private) in Perth, Western Australia. We recruited 210 adults aged 50 years or older who were due to undergo coronary artery bypass grafting or valve replacement surgery. Participants were randomly assigned (1:1) to 7 days of treatment with melatonin 3 mg at night or matching placebo, starting 2 days before the surgery. The primary outcome of interest was incident delirium within 7 days of surgery as assessed via daily clinical assessment that included the Confusion Assessment Method. Secondary outcomes of interest included duration and severity of delirium, length of hospital stay, cognitive function, and mood and anxiety symptoms at discharge and 3 months after the surgery. The groups were well balanced for demographic and clinical parameters. Forty-two participants developed delirium, but it was evenly distributed between the groups (melatonin 21/98, 21.4%; placebo 21/104, 20.2%; adjusted odds ratio [OR] = .78; 95% confidence interval [CI] = .35-1.75). The median duration of delirium was 3 (interquartile range [IQR] = 2-4) and 2 (IQR = 1-3) days for people treated with melatonin and placebo, respectively (z = -1.03; P = .304). A similar proportion of participants experienced severe episodes of delirium in each group (melatonin 9/21, 42.9% vs placebo 6/21, 28.6%; χ2 = .93; P = .334; adjusted OR = 1.98; 95% CI = .40-9.78). The groups did not differ in terms of length of stay, mood, anxiety, and cognitive performance. The findings of this randomized double-blind placebo-controlled trial do not support the prophylactic use of melatonin to prevent delirium after major cardiac surgery. J Am Geriatr Soc 68:112-119, 2019.


Noncontact holmium:YAG laser thermal keratoplasty to correct hyperopia: 18-month follow-up.
To assess the safety and efficacy of noncontact holmium:yttrium aluminium garnet laser thermal keratoplasty (Ho:YAG LTK) for correction of low to moderate hyperopia. We performed noncontact Ho:YAG LTK on 1 eye each of 28 patients for correction of hyperopia up to +3.88 dpt. Treatments were conducted with 1 or 2 symmetrical octagonal rings of 8 spots/ring with centerline diameters of 6 mm (1 ring) or 6 and 7 mm (2 rings), 10 pulses of laser light at 5 Hz pulse repetition frequency, variable pulse energy in the range of 208-242 mJ and a nominal spot diameter between 615 and 623 microns. At 18 months after surgery, 20 of 22 (91%) treated patient eyes had improved uncorrected distance visual acuity. The mean change in subjective manifest refraction (spherical equivalent) was -0.52 +/- 0.35 dpt and -1.41 +/- 0.53 dpt for 1- and 2-ring treatment groups, respectively, with good stability in the refractive change after 6 months. The mean induced refractive astigmatism was small (0.30 +/- 0.37 dpt/0.25 +/- 0.29 dpt for 1-/2-ring treatments). None of the eyes lost 2 or more lines of spectacle-corrected distance visual acuity. There were no clinically significant changes in glare and contrast sensitivity. Noncontact LTK treatment of low hyperopia is safe and effective, and it is more stable and less prone to induce astigmatism than previously reported contact mode LTK treatments.


Riding the wave of innovation: immunoinformatics in fish disease control.
The spread of infectious illnesses has been a significant factor restricting aquaculture production. To maximise aquatic animal health, vaccination tactics are very successful and cost-efficient for protecting fish and aquaculture animals against many disease pathogens. However, due to the increasing number of immunological cases and their complexity, it is impossible to manage, analyse, visualise, and interpret such data without the assistance of advanced computational techniques. Hence, the use of immunoinformatics tools is crucial, as they not only facilitate the management of massive amounts of data but also greatly contribute to the creation of fresh hypotheses regarding immune responses. In recent years, advances in biotechnology and immunoinformatics have opened up new research avenues for generating novel vaccines and enhancing existing vaccinations against outbreaks of infectious illnesses, thereby reducing aquaculture losses. This review focuses on understanding in silico epitope-based vaccine design, the creation of multi-epitope vaccines, the molecular interaction of immunogenic vaccines, and the application of immunoinformatics in fish disease based on the frequency of their application and reliable results. It is believed that it can bridge the gap between experimental and computational approaches and reduce the need for experimental research, so that only wet laboratory testing integrated with in silico techniques may yield highly promising results and be useful for the development of vaccines for fish.


[Psychologic adjustment to therapy in children with cancer].
Peculiarities of adjustment of pediatric cancer patients and their perception of the disease were studied in a group of 29 including 12 cases of hematooncological diseases and 17 cases of different solid tumors. Projective psychological methods were used. Such menifestations of failure to adjust as lower selfesteem, feeling of guilt, helplessness, passivity, anxiety and fear were observed in nearly all the patients. Adequate adjustment was found to be determined by the formation of a similarly adequate perception of the disease which in turn becomes possible when the problems involved were discussed with the patient by competent medical staff and well-counseled parents in an open and trust-worthy manner.


Bridle procedure combined with a subtalar implant: a case series and review of the literature.
The bridle procedure is a tritendon anastomosis between the tibialis posterior, peroneus longus, and tibialis anterior, utilized in the treatment of drop foot. Concerned about the potential for acquired flatfoot deformity following the bridle procedure, the authors hypothesized that placing a sinus tarsi implant would prevent this potential sequela. Over a 10-year period, 15 patients (16 feet) were treated with a bridle procedure for drop foot, and 9 of these feet also had a subtalar arthroereisis implanted; 8 patients (9 feet) positively responded to requests for long-term follow-up evaluation and returned for subjective evaluation with the American Orthopaedic Foot and Ankle Society (AOFAS) Hindfoot Scale and Short Form (SF)-36 forms and objective clinical and radiographic measurements. At an average follow-up of 61 months, the 8 patients (9 feet) who returned for long-term evaluation maintained a plantigrade foot, were able to dorsiflex above neutral both actively and passively, and did not develop a flatfoot deformity at long-term follow-up. There were no significant differences between those who had a sinus tarsi implant (6 feet) and those who did not (3 feet). Although 7 of the 9 feet evaluated in this study wore a brace preoperatively, none continued to use a brace after surgery. The major complaints were occasional pain at the subtalar arthroereisis site and a feeling that the great toe was dragging. The bridle procedure provided excellent outcomes in patients with drop foot, with patients achieving long-term success in being free from brace wear. The ability to actively dorsiflex the foot during gait was also restored, reestablishing a normal heel-to-toe gait pattern. Therapeutic, Level IV: Case series.


In vitro effects of Warburgia ugandensis, Psiadia punctulata and Chasmanthera dependens on Leishmania major promastigotes.
Plant extracts from Warburgia ugandensis Sprague (Family: Canellaceae), Psiadia punctulata Vatke (Family: Compositae) and Chasmanthera dependens Hoschst (Family: Menispermaceae) were tested for activity on Leishmania major promastigotes (Strain IDU/KE/83 = NLB-144) and infected macrophages in vitro. Plants were collected from Baringo district, dried, extracted, weighed and tested for antileishmanial activity. Serial dilutions of the crude extracts were assayed for their activity against Leishmania major in cell free cultures and in infected macrophages in vitro. Inhibitory concentrations and levels of cytotoxicity were determined. Warburgia ugandensis, Psiadia punctulata and Chasmanthera dependens had an IC(50) of 1.114 mg/ml, 2.216 mg/ml and 4.648 mg/ml, respectively. The cytotoxicity of the drugs on BALB/c peritoneal macrophage cells was insignificant as compared to the highly toxic drug of choice Pentostam(®). The supernatants from control and Leishmania infected macrophages were analyzed for their nitrite contents by Griess reaction and nitrite absorbance measured at 540 nm. Warburgia ugandensis (stem bark water extract), Chasmanthera dependens (stem bark water extract) and Psiadia punctulata (stem bark methanol extract) produced 112.3%, 94% and 88.5% more nitric oxide than the untreated infected macrophages respectively. Plant crude extracts had significant (p<0.05) anti-leishmanial and immunomodulative effects but insignificant cytotoxic effects at 1mg/ml concentration. All experiments were performed in triplicate. Statistical analysis of the differences between mean values obtained from the experimental group compared to the controls was done by students't test. ANOVA was used to determine the differences between the various treatment groups. The analysis program Probit was used to determine IC(50)s.


Immunomonitoring of Monocyte and Neutrophil Function in Critically Ill Patients: From Sepsis and/or Trauma to COVID-19.
Immune cells and mediators play a crucial role in the critical care setting but are understudied. This review explores the concept of sepsis and/or injury-induced immunosuppression and immuno-inflammatory response in COVID-19 and reiterates the need for more accurate functional immunomonitoring of monocyte and neutrophil function in these critically ill patients. in addition, the feasibility of circulating and cell-surface immune biomarkers as predictors of infection and/or outcome in critically ill patients is explored. It is clear that, for critically ill, one size does not fit all and that immune phenotyping of critically ill patients may allow the development of a more personalized approach with tailored immunotherapy for the specific patient. In addition, at this point in time, caution is advised regarding the quality of evidence of some COVID-19 studies in the literature.


Revealing the adsorption mechanisms of nitroxides on ultrapure, metallicity-sorted carbon nanotubes.
Carbon nanotubes are a natural choice as gas sensor components given their high surface to volume ratio, electronic properties, and capability to mediate chemical reactions. However, a realistic assessment of the interaction of the tube wall and the adsorption processes during gas phase reactions has always been elusive. Making use of ultraclean single-walled carbon nanotubes, we have followed the adsorption kinetics of NO2 and found a physisorption mechanism. Additionally, the adsorption reaction directly depends on the metallic character of the samples. Franck-Condon satellites, hitherto undetected in nanotube-NOx systems, were resolved in the N 1s X-ray absorption signal, revealing a weak chemisorption, which is intrinsically related to NO dimer molecules. This has allowed us to identify that an additional signal observed in the higher binding energy region of the core level C 1s photoemission signal is due to the C ═ O species of ketene groups formed as reaction byproducts . This has been supported by density functional theory calculations. These results pave the way toward the optimization of nanotube-based sensors with tailored sensitivity and selectivity to different species at room temperature.


Tracing the Mediterranean diet through principal components and cluster analyses in the Greek population.
To identify dietary patterns, and their socio-demographic and lifestyle correlates in a large sample of Greek adults, and assess their adherence to the traditional Mediterranean diet. Principal component (PC) analysis was used to identify dietary patterns among 28 034 participants of the Greek branch of the European Prospective Investigation into Cancer and Nutrition. Dietary information was collected through a validated, semiquantitative, food-frequency questionnaire. The extracted PCs were subsequently regressed on sociodemographic and lifestyle variables. Analyses were also performed to classify individuals with similar dietary behavior into clusters. Four PCs were identified: PC1 resembled the Mediterranean diet, PC2 approximated a vegetarian diet with emphasis on seed oils, PC3 reflected a preference for sweets, and PC4 reflected a Western diet. PC1 and PC2 were positively associated with age, education, physical activity, and nonsmoking status. Females, in comparison to males, scored higher on PC1 but lower on PC2. Males, younger, more educated individuals, nonsmokers and residents of Greater Athens (Attica) scored higher on PC3. PC4 was associated with younger age, less education, and current smoking. In cluster analyses, cluster A contrasted clusters B and C in having much higher mean PC1- and PC2-scores and substantially lower PC3- and PC4-scores. PC1 and PC4 were, respectively, positively and inversely correlated with an a priori Mediterranean-diet score; PC2 and PC3 were unrelated to it. The Mediterranean-like PC1-score as well as the vegetarian-like PC2 were higher among older, more educated people, and were associated with a healthier lifestyle than PC4, which reflected a Western-type diet. PC1 was strongly positively associated with an a priori Mediterranean-diet score.


Hypermutation rate normalized by chronological time.
It is generally believed that in cells undergoing Ig somatic hypermutation, more cell divisions result in more mutations. This is because DNA synthesis and replication is thought to play roles in the known mechanisms-cytidine deamination and subsequent conversion to thymidine, uracil-DNA glycosylase-mediated repair, mismatch repair, and DNA synthesis by error-prone polymerases. In this study, we manipulated the number of cell generations by varying the rate at which cultures of a mouse cell line were replenished with fresh medium. We found that the frequency of mutants does not necessarily increase with the number of cell generations. On the contrary, a greater number of divisions can lead to a lower frequency of mutants, indicating that cell division is not a rate-limiting step in the hypermutation process. Thus, when comparing mutation rates, we suggest that rates are more appropriately expressed as mutations per day than per cell generation.


[Site-directed mutagenesis of sugar-binding lectin fragments of legume plants with the help of inverse-PCR].
Method of site-directed mutagenesis that gave an opportunity to the site-independent manipulation with enough extended exposed mutagenesis fragments of gene cloning in plasmid vector was developed. With the help of this method hybrid constructions representing peas Pisum sativum (PSL) lectine with carbohydrate-binding fragment changing on that lectines from another legume plants were created. This method is suitable for obtaining the expression constructions in plasmid vectors, when is necessary to put a coding part under the control of promoter in a certain reading frame.


Considerations in Planning Physical Activity for Older Adults in Hot Climates: A Narrative Review.
Regular physical activity has multiple health benefits for both the prevention and management of disease, including for older adults. However, additional precautions are needed with ageing given physiological changes and the increasing prevalence of comorbidities. Hot ambient temperatures increase the risks of exercise at any age, but are particularly important given thermoregulatory changes in older people. This narrative review informs planning of physical activity programs for older people living in rural areas with very hot climates for a period of the year. A multi-database search of peer-reviewed literature was undertaken with attention to its relevance to Australia, starting with definitions and standard advice in relation to physical activity programming and the incremental limitations imposed by age, rurality, and extreme heat. The enablers of and barriers to increasing physical activities in older adults and how they can be modified for those living in extreme hot climates is described. We describe multiple considerations in program design to improve safety, adherence and sustaining physical activity, including supervision, simple instructions, provision of reminders, social support, encouraging self-efficacy. Group-based activities may be preferred by some and can accommodate special populations, cultural considerations. Risk management is an important consideration and recommendations are provided to assist program planning.


Role of extra- and intracellular Ca2+ in the lymphatic myogenic response.
We used an actively contracting in vitro preparation of bovine mesenteric lymph vessels to study the effect of selected calcium channel modulators on the ability of these vessels to propel fluid. We found that blocking the dihydropyridine receptor with nifedipine and diltiazem inhibited pumping at concentrations within the range used clinically (10(-7) and 10(-6) M, respectively). Intracellular calcium modulation using ryanodine (10(-6) M) also inhibited pumping. Furthermore, we studied the effect of these agents on the relationship between lymph flow and transmural pressure, a relationship normally described by a bell-shaped curve. Diltiazem, 10(-6) M, attenuated pumping over the range of pressures studied. On the other hand, ryanodine at 10(-7) M, a concentration capable of inhibiting pumping at a constant transmural pressure, had no effect on the pressure-flow relationship when transmural pressure was manipulated. Thus we have determined that calcium movement via the L-type channel contributes significantly to the regulation of lymph pump activity and that intracellular calcium flux plays a less significant role that may be modified by transmural pressure.


[Recent genetic studies on manic-depression: evaluation and methodology].
In this article, the part of hereditary factors in the onset of depressive diseases is analysed. Primary depressive diseases may be now separated in two groups: one includes bipolar depressive syndromes. (e.i. of manic depressive type) and the other the monopolar depressive syndromes (e.i. of depressive type). A short review of twin studies and family studies in the manic depressive type bipolar depressive illnesses is made. From these studies, a discussion about genetical hypothesis concerning the manic depressive illnesses is proposed. Different genetical hypotheses are revisited, and that of a dominant X-linked disease is observed. Genetical factors are also revisited into the heterogeneous group of monopolar depressive diseases, emphasizing the different depressive subgroups of this large category. Finally, the authors also consider the possible relationship between bipolar manic depressive syndromes and schizophrenia, trying to classify the dysthymic schizophrenia.


Proteomic Characterisation of Lupin (Lupinus angustifolius) Milk as Influenced by Extraction Techniques, Seed Coat and Cultivars.
Lupin seeds are rich in proteins and other essential ingredients that can help to improve human health. The protein contents in both whole and split seeds of two lupin cultivars (Mandleup and PBA Jurien) were used to produce the lupin milk using the cheesecloth and centrifuge method. Proteins were extracted from the lupin milk using thiourea/urea solubilization. The proteins were separated by a two-dimensional polyacrylamide gel electrophoresis and then identified with mass spectrometry. A total of 230 protein spots were identified, 60 of which showed differential abundances. The cheesecloth separation showed protein extractability much better than that of the centrifuge method for both the cultivars. The results from this study could offer guidance for future comparative analysis and identification of lupin milk protein and provide effective separation technique to determine specific proteins in the cheese-making process.


Liposome-incorporated santolina insularis essential oil: preparation, characterization and in vitro antiviral activity.
The effect of liposomal inclusion on the stability and in vitro antiherpetic activity of Santolina insularis essential oil was investigated. In order to study the influence of vesicle structure on the liposome properties, multilamellar and unilamellar vesicles were prepared by the film method and sonication, respectively. Vesicles were obtained from hydrogenated soya phosphatydilcholine and cholesterol. Formulations were examined for their stability for over one year monitoring the drug leakage from vesicles and the average size distribution. The stability of the incorporated oil was verified by studying its quali-quantitative composition. The antiviral activity was studied against Herpes simplex virus type 1 (HSV-1) by plaque reduction and yield reduction assays. Results showed that Santolina insularis essential oil can be incorporated in high amounts in the prepared liposomes, which successfully prevented its degradation. Moreover, stability studies pointed out that vesicle dispersions were stable for at least one year and neither oil leakage nor vesicle size alteration occurred during this period. Antiviral activity assays demonstrated that Santolina insularis essential oil is effective in inactivating HSV-1 and that the activity is principally due to direct virucidal effects. Free essential oil proved to be more effective than liposomal oil and a different activity was discovered which related to the vesicular structure. The ED(50) values, significantly lower when cells were pre-incubated with the essential oil before the virus adsorption, indicate an intracellular mechanism in the antiviral activity of Santolina insularis. Moreover, liposomal Santolina essential oil is non toxic in the range of the concentration tested.


HIV-1 induces B-cell activation and class switch recombination via spleen tyrosine kinase and c-Jun N-terminal kinase pathways.
Patients infected by the HIV type 1 (HIV-1) frequently show a general deregulation of immune system. A direct influence of HIV-1 particles on B-cell activation, proliferation and B-cell phenotype alterations has been recently described. Moreover, expression of activation-induced cytidinedeaminase (AID) mRNA, which is responsible for class switch recombination (CSR) and somatic hypermutation (SHM), was reported to be overexpressed in B cells exposed to HIV-1. Study of primary human B cells in an in-vitro model. In the current study, we evaluated which signalling pathways are activated in primary B cells after a direct contact with HIV-1 particles in vitro using different kinase inhibitors. Here, we report that B-cell activation together with the increase of AID mRNA expression and the subsequent class switch recombination (CSR) in HIV-exposed B cells occurred through spleen tyrosine kinase (SYK) and c-Jun N-terminal kinase (JNK) pathways. Therefore, we showed that HIV-1 could directly induce primary B-cell deregulation via SYK/B-cell receptor (BCR) engagement, and that activation was followed by the JNK pathway activation. To our knowledge, these data provide the first evidence that SYK/BCR activation was the first step for B-cell activation and CSR mechanism after HIV-1 stimulation in a T-cell-free context.


Loss of heterozygosity for the short arm of chromosome 11 (11p15) in human milk epithelial cells immortalized by microinjection of SV40 DNA.
We have developed, by microinjection of SV40 DNA into human milk epithelial cells, a new mammary cell line, Hu-MI, which exhibits the phenotype of luminal cells or so-called "breast cancer precursor cells." This cell line retains the phenotype of primary cells as demonstrated by the expression of keratins 18 and 19 and of polymorphic epithelial mucins. However, the cells do not grow in agar after more than 80 passages, nor do they form tumors in nude mice. Established cells contain 2 copies of SV40 DNA integrated into the cellular genome and up to 14 copies of free SV40 DNA. A deletion of the short arm of chromosome 11 (11p15) including the c-Ha-ras and the beta-globin genes was found in the immortalized cells when the DNA from these cells was compared to the DNA from peripheral blood mononuclear cells obtained from the same donor. In addition, this cell line showed a good transfection efficiency for other DNA sequences using classical transfection and selection techniques with a neomycin resistance gene (pKOneo). Selective microinjection of DNA into tumor precursor cells may prove useful for the study of the molecular mechanisms involved in breast carcinogenesis. The possible significance of the loss of 11p13-15 in malignant progression of breast cancer is discussed.


Pancreatic alpha cell-selective deletion of Tcf7l2 impairs glucagon secretion and counter-regulatory responses to hypoglycaemia in mice.
Transcription factor 7-like 2 (TCF7L2) is a high mobility group (HMG) box-containing transcription factor and downstream effector of the Wnt signalling pathway. SNPs in the TCF7L2 gene have previously been associated with an increased risk of type 2 diabetes in genome-wide association studies. In animal studies, loss of Tcf7l2 function is associated with defective islet beta cell function and survival. Here, we explore the role of TCF7L2 in the control of the counter-regulatory response to hypoglycaemia by generating mice with selective deletion of the Tcf7l2 gene in pancreatic alpha cells. Alpha cell-selective deletion of Tcf7l2 was achieved by crossing mice with floxed Tcf7l2 alleles to mice bearing a Cre recombinase transgene driven by the preproglucagon promoter (PPGCre), resulting in Tcf7l2AKO mice. Glucose homeostasis and hormone secretion in vivo and in vitro, and islet cell mass were measured using standard techniques. While glucose tolerance was unaffected in Tcf7l2AKO mice, glucose infusion rates were increased (AUC for glucose during the first 60 min period of hyperinsulinaemic-hypoglycaemic clamp test was increased by 1.98 ± 0.26-fold [p < 0.05; n = 6] in Tcf7l2AKO mice vs wild-type mice) and glucagon secretion tended to be lower (plasma glucagon: 0.40 ± 0.03-fold vs wild-type littermate controls [p < 0.01; n = 6]). Tcf7l2AKO mice displayed reduced fasted plasma glucose concentration. Glucagon release at low glucose was impaired in islets isolated from Tcf7l2AKO mice (0.37 ± 0.02-fold vs islets from wild-type littermate control mice [p < 0.01; n = 6). Alpha cell mass was also reduced (72.3 ± 20.3% [p < 0.05; n = 7) in Tcf7l2AKO mice compared with wild-type mice. The present findings demonstrate an alpha cell-autonomous role for Tcf7l2 in the control of pancreatic glucagon secretion and the maintenance of alpha cell mass and function.


Perfectionism and training performance: The mediating role of other-approach goals.
Recent research found perfectionistic strivings to predict performance in a novel basketball task among novice basketball players. The current study builds on this research by examining whether this is also the case for performance in a familiar basketball training task among experienced basketball players, and whether achievement goals mediated any observed relationships. Perfectionistic strivings, perfectionistic concerns, and 3 × 2 achievement goals were assessed prior to basketball training performance in 90 basketball players (mean age 20.9 years). Regression analyses showed that perfectionistic strivings predicted better performance. Furthermore, mediation analyses showed that other-approach goals (e.g. beliefs that one should and can outperform others) accounted for this relationship. The findings suggest that perfectionistic strivings may predict better performance in both novel and familiar athletic contexts. In addition, beliefs about the importance and ability to outperform others may explain this relationship.


Peripheral arylation of subporphyrazines.
Peripherally hexaarylated subporphyrazines (SubPzs) have been prepared through a Pd-catalyzed, CuTC-mediated coupling of a hexaethylsulfanylated subporphyrazine with arylboronic acids. The introduced aryl substituents strongly influence the electronic properties of the subporphyrazine through effective conjugative interaction. Aryl rings endowed with π-electron-donating groups at the para positions produce a remarkable perturbation of the electron density of the SubPz macrocycle. This is reflected through significant redshifts of the SubPz CT and Q-bands, together with increase of the molar absorptivity of the former, with respect to those exhibited by the hexaphenyl-SubPz 2 a. Moreover, the trend in the first SubPz reduction potentials correlates with the Hammett constants (σp ) corresponding to the para substituents of the aryl. The domed, extended SubPz π-system self-assembles in the solid state to form a dimeric capsule that houses a solvent molecule.


Focal left atrial tachycardia in a patient with left ventricular noncompaction.
Left ventricular noncompaction (LVNC) is a rare disease caused by intrauterine failure of the myocardium to compact. The major clinical manifestations of LVNC include heart failure, ventricular tachyarrhythmia, thromboembolic event, and sudden deaths. Atrial arrhythmia usually seen is atrial fibrillation. We report a rare case of focal left atrial tachycardia in an 18-year-old patient who presented for evaluation of persistent tachycardia. Transthoracic echocardiogram showed severe systolic dysfunction and evidence of noncompaction of the left ventricle. A detailed review of ECG revealed the possibility of ectopic atrial tachycardia, most likely originating from the left side. Electrophysiology study showed sustained atrial tachycardia originating on the ridge anterior to the left sided pulmonary veins. A successful radiofrequency catheter ablation was performed at this site without any complications.


[Control and regulation of the blood circulation in the vertebro-basilar system. Review of the literature].
The author establishes an analogy between the control mechanism and regulation of the cerebral blood flow and of protection of the vascular wall of the internal carotid constituted by the conjunction "internal carotid-cavernous sinus" with the group represented by the system "vertebro basilar-transverse occipital sinus or basilar" (an extension of the cavernous sinus) in the regulation and control of the encephalic circulation carried out through this latter vessels, together with the protection of its vascular walls. The author believes to be very difficult to demonstrate in practice the functioning of these mechanisms, but it is very logical and easy to reason about them, to value them and to give the importance and meaning or motive which forcibly they should have and not simply consider them as freaks of nature and bizarre anatomical features.


Phosphoproteomics profiling of human skin fibroblast cells reveals pathways and proteins affected by low doses of ionizing radiation.
High doses of ionizing radiation result in biological damage; however, the precise relationships between long-term health effects, including cancer, and low-dose exposures remain poorly understood and are currently extrapolated using high-dose exposure data. Identifying the signaling pathways and individual proteins affected at the post-translational level by radiation should shed valuable insight into the molecular mechanisms that regulate dose-dependent responses to radiation. We have identified 7117 unique phosphopeptides (2566 phosphoproteins) from control and irradiated (2 and 50 cGy) primary human skin fibroblasts 1 h post-exposure. Semi-quantitative label-free analyses were performed to identify phosphopeptides that are apparently altered by radiation exposure. This screen identified phosphorylation sites on proteins with known roles in radiation responses including TP53BP1 as well as previously unidentified radiation-responsive proteins such as the candidate tumor suppressor SASH1. Bioinformatic analyses suggest that low and high doses of radiation affect both overlapping and unique biological processes and suggest a role for MAP kinase and protein kinase A (PKA) signaling in the radiation response as well as differential regulation of p53 networks at low and high doses of radiation. Our results represent the most comprehensive analysis of the phosphoproteomes of human primary fibroblasts exposed to multiple doses of ionizing radiation published to date and provide a basis for the systems-level identification of biological processes, molecular pathways and individual proteins regulated in a dose dependent manner by ionizing radiation. Further study of these modified proteins and affected networks should help to define the molecular mechanisms that regulate biological responses to radiation at different radiation doses and elucidate the impact of low-dose radiation exposure on human health.


Uraemic Cardiomyopathy: A Review of Current Literature.
Uraemic Cardiomyopathy (UC) is recognised as an intricate and multifactorial disease which portends a significant burden in patients with End-Stage Renal Disease (ESRD). The cardiovascular morbidity and mortality associated with UC is significant and can be associated with the development of arrythmias, cardiac failure and sudden cardiac death (SCD). The pathophysiology of UC involves a complex interplay of traditional implicative factors such as haemodynamic overload and circulating uraemic toxins as well as our evolving understanding of the Chronic Kidney Disease-Mineral Bone Disease pathway. There is an instrumental role for multi-modality imaging in the diagnostic process; including transthoracic echocardiography and cardiac magnetic resonance imaging in identifying the hallmarks of left ventricular hypertrophy and myocardial fibrosis that characterise UC. The appropriate utilisation of the aforementioned diagnostics in the ESRD population may help guide therapeutic approaches, such as pharmacotherapy including beta-blockers and aldosterone-antagonists as well as haemodialysis and renal transplantation. Despite this, there remains limitations in effective therapeutic interventions for UC and ongoing research on a cellular level is vital in establishing further therapies.


The effect of image descriptors on the performance of classifiers of footwear outsole image pairs.
Shoe prints are commonly found at the scene of a crime and can sometimes help link a suspect to the scene. Because prints tend to be partially observed or smudgy, comparing crime scene prints with reference images from a putative shoe can be challenging. Footwear examiners rely on guidelines such as those published by SWGTREAD [1] to visually assess the similarity between two or more footwear impressions, one reason being that reliable, quantitative methods have yet to be validated for use in real cases. To help in the development of such methods, we created a study dataset of images of outsole impressions that shared class characteristics and degree of wear and that were subject to a specific type of degradation. We also propose a method to quantify the similarity between two outsole images that extends the capabilities of MC-COMP [2]. The proposed method is composed of three steps; (1) extracting image descriptors, (2) aligning images using the maximum clique, (3) calculating similarity values using two different classifiers; (a) degree of overlap between the two images, and (b) a score produced by a random forest. To explore the performance of the algorithm we propose, we compared degraded, crime scene-like images to high-quality reference images produced by the same or by different shoes. Even though comparisons involved matches or very close non-matches, and one of the images was blurry, the algorithm shows good source classification performance.


Determinants of Improvement in Cardiopulmonary Exercise Testing After Left Ventricular Assist Device Implantation.
Although improvement in cardiac output and hemodynamic parameters is routinely demonstrated in patients implanted with continuous-flow left ventricular assist devices (CF-LVADs), improvement in exercise capacity is inconsistently seen. Our purpose was to determine whether native cardiac factors, LVAD factors, or comorbid factors were associated with lack of improvement. Review of all patients undergoing preimplant cardiopulmonary exercise testing (CPET) and CPET on LVAD therapy at one institution was performed between 2007 and 2014 (n = 49). Comprehensive assessment of echocardiographic parameters, right heart catheterization data, medications, and comorbid illness was undertaken. There was no mean improvement in peak oxygen consumption (VO2; 11.8-12.4 ml/kg/min; p = 0.26), although exercise time (5.1 [46% predicted] to 5.8 min [56% predicted]; p = 0.02) and nadir of the ratio of minute ventilation to carbon dioxide production slope (VE/VCO2; 39-36; p = 0.001) improved. Factors most strongly associated with improvement in VO2 were Heartmate II pulsatility index (PI; R = 0.48; p = 0.001), power (R = -0.40; p = 0.009), pump flow (R = -0.40; p = 0.008), and pump speed (R = -0.32; p = 0.04). Peak heart rate (HR) was also associated with improvement in VO2 (R = 0.41; p = 0.004). Left ventricular ejection fraction (LVEF; R = 0.004; p = 0.77), right ventricular (RV) function (R = 0.22; p = 0.28), and aortic valve opening (R = 0.20; p = 0.57) were not associated with improvement in VO2. Our data suggest that less reliance on LVAD support was modestly associated with improvement in exercise capacity. Further studies should seek to determine the optimal level of device support prospectively in relation to exercise capacity.


An intact sialoadhesin (Sn/SIGLEC1/CD169) is not required for attachment/internalization of the porcine reproductive and respiratory syndrome virus.
Surface expression of SIGLEC1, also known as sialoadhesin or CD169, is considered a primary determinant of the permissiveness of porcine alveolar macrophages for infection by porcine reproductive and respiratory syndrome virus (PRRSV). In vitro, the attachment and internalization of PRRSV are dependent on the interaction between sialic acid on the virion surface and the sialic acid binding domain of the SIGLEC1 gene. To test the role of SIGLEC1 in PRRSV infection, a SIGLEC1 gene knockout pig was created by removing part of exon 1 and all of exons 2 and 3 of the SIGLEC1 gene. The resulting knockout ablated SIGLEC1 expression on the surface of alveolar macrophages but had no effect on the expression of CD163, a coreceptor for PRRSV. After infection, PRRSV viremia in SIGLEC1(-/-) pigs followed the same course as in SIGLEC1(-/+) and SIGLEC1(+/+) littermates. The absence of SIGLEC1 had no measurable effect on other aspects of PRRSV infection, including clinical disease course and histopathology. The results demonstrate that the expression of the SIGLEC1 gene is not required for infection of pigs with PRRSV and that the absence of SIGLEC1 does not contribute to the pathogenesis of acute disease.


The MAM (meprin/A5-protein/PTPmu) domain is a homophilic binding site promoting the lateral dimerization of receptor-like protein-tyrosine phosphatase mu.
The MAM (meprin/A5-protein/PTPmu) domain is present in numerous proteins with diverse functions. PTPmu belongs to the MAM-containing subclass of protein-tyrosine phosphatases (PTP) able to promote cell-to-cell adhesion. Here we provide experimental evidence that the MAM domain is a homophilic binding site of PTPmu. We demonstrate that the MAM domain forms oligomers in solution and binds to the PTPmu ectodomain at the cell surface. The presence of two disulfide bridges in the MAM molecule was evidenced and their integrity was found to be essential for MAM homophilic interaction. Our data also indicate that PTPmu ectodomain forms oligomers and mediates the cellular adhesion, even in the absence of MAM domain homophilic binding. Reciprocally, MAM is able to interact homophilically in the absence of ectodomain trans binding. The MAM domain therefore contains independent cis and trans interaction sites and we predict that its main role is to promote lateral dimerization of PTPmu at the cell surface. This finding contributes to the understanding of the signal transduction mechanism in MAM-containing PTPs.


Prospective Multicenter Experience With Cooled Radiofrequency Ablation Using High Impedance Irrigant to Target Deep Myocardial Substrate Refractory to Standard Ablation.
This study sought to evaluate the efficacy and safety of using half-normal saline (HNS) as the cooling radiofrequency ablation (RFA) irrigant among patients who had failed prior, standard RFA. Effective control of ventricular arrhythmias that arise from mid-myocardium may be refractory to standard RFA. Recent data suggest that delivering fluid with decreased ionic concentration during open-irrigated RFA can produce deeper RFA lesions. A 12-center prospective analysis was performed of all ablations using HNS for the treatment of ventricular arrhythmias (premature ventricular complex [PVC]/ventricular tachycardia [VT]) refractory to standard ablation with normal saline irrigant. HNS RFA was used clinically to target 94 PVC/VTs refractory to standard ablation. Acute success was achieved in 78 of 94 (83%), with longer-term success occurring in 78 subjects after a mean follow-up of 6.1 ± 6.7 months (range, 3.0 to 25.2 months). Steam pops were observed among 12 (12.6%) patients. There were no significant changes in electrolytes measured before and after the use of HNS, and there were no complications related to HNS use. The use of HNS instead of normal saline irrigant during high-power delivery targeting deep myocardial substrate is safe and effective. PVC/VT sources previously unaffected by standard ablation may be successfully ablated with improved efficiency of radiofrequency delivery using HNS.


Incidence of Anal Dysplasia in a Population of High-Risk Women: Observations at a Cervical Pathology Unit.
This study aimed to evaluate the incidence of anal dysplasia in women at high risk of developing those lesions and to assess the relationships between positive anal cytology and different risk factors. We performed an observational cross-sectional study involving a patient survey and chart review. The study was conducted at the cervical pathology unit at the Hospital Universitario 12 de Octubre, Madrid, Spain, from 2011 to 2015. Patients were 215 women aged 18 to 65 years old with risk factors for anal dysplasia, for whom anal evaluation was indicated. Anal cytology was performed in all patients. High-resolution anoscopy and anal biopsy were used to investigate abnormalities. All patients completed a survey regarding sexual practices. The patients' demographic and clinical data were collected by using a retrospective chart review. The main outcome measure was the incidence of anal dysplasia in this population. Of the 215 patients, 45 (21.0%) presented with cytological abnormalities (atypical squamous cells of undetermined significance, 13.5%; low-grade squamous intraepithelial lesion, 5.6%; high-grade squamous intraepithelial lesion, 1.9%). Anoscopy was performed in 31 patients (14.4%) and 2 patients (0.9%) had abnormal findings. One case (0.5%) of low-grade anal intraepithelial neoplasia was identified. Only immunosuppression (p = .01) and smoking status (p = .02) were significantly correlated with positive anal cytology results. Limitations of the study include the small single-center sample, a lack of controls, the retrospective design, potential survey response biases, and the nonstandardized survey. The incidence of abnormal cytological findings was 21.0%, whereas low-grade anal intraepithelial was confirmed in 0.5% of the patients. There is a higher incidence of cytological abnormalities among immunosuppressed women and smokers.


Stable isotope ratiometer-multiple ion detector unit for quantitative and qualitative stable isotope studies by gas chromatography-mass spectrometry.
We have designed and constructed a stable isotope ratiometer-multiple ion detector unit, which can drive existing chromatograph-quadrupole or magnetic sector mass gas spectrometers to monitor up to six ions in turn. Each of the three pairs of ions can be selected for quantitation; thus three different or successive components can be analyzed in a single gas-chromatographic run. A background subtraction option permits the ion intensity in the absence of sample to be subtracted automatically during sample measurement. Displays of accumulated counts and isotope ratio are updated twice per second during the measurement and can be printed out at is conclusion. All six ions can be monitored in the analog mode by parallel outputs to a multipen recorder. Experience gained in the construction of this prototype indicates that such units could be commercially available for $10 000, or about a third to a sixth of the cost of even an inexpensive computer system.


Low frequency of the ccr5delta32 HIV-resistance allele in mainland China: identification of the first case of ccr5delta32 mutation in the Chinese population.
A 32-bp deletion on the CCR5 gene (ccr5delta32) confers resistance to HIV-1 infection. This deletion is common in Caucasians, but rare in Asians. Since the frequency of the ccr5delta32 allele of Chinese in mainland China has been unknown we investigated the ccr5delta32 mutation in a cohort of 407 Chinese people in this area. A 225-bp fragment of CCR5 encompassing the 32-bp region was analysed by PCR, hybridization and sequencing. Only 1 out of 407 subjects was heterozygous for ccr5delta32 and no homozygotes were detected. The frequency of ccr5delta32 in this cohort is thus 0.00123, i.e. much lower than that of Caucasians. The ccr5delta32 heterozygote is a healthy young man. To our knowledge this is the first ccr5delta32 mutant found in Chinese people. The results indicate that ccr5delta32 does exist in Chinese people, but at very low frequency. This suggests that ccr5delta32 is not a significant factor for the genetic resistance to HIV-1 in Chinese people.


Complement activation reaches maximum during equilibrium between antigen and antibody in an in vitro model for thrombolysis with streptokinase.
Treatment with streptokinase (SK) of patients with acute myocardial infarction (AMI) is known to activate the complement system. In a previous investigation we found a great variation in the degree of complement activation taking place during infusion of SK in patients with AMI (range: 0-293% increase in pretreatment complement split product 3d (C3d) levels). In this in vitro study we added SK to serum with different concentrations of SK-antibodies (SK-Ab) and found that both the relative concentrations and the total concentrations of SK and Ab determined the degree of complement activation elicited by SK. This finding was in accordance with the Kendall-Heidelberger equation of antigen-antibody equivalence. Furthermore, we found parallel rises in C3d and C4d, indicating that complement was activated via the classical pathway. A condition for maximal complement activation is antigen-antibody equivalence.


Assessment of selected heavy metals pollution in water, sediments and fish in the basin Dyje, Czech Republic.
This study presents the concentrations of heavy metals (Cr, Cd, Hg, Zn and Pb) in the water, sediment, and muscle of two fish species from the middle and lower reaches of the Dyje river basin (Dyje, Jihlava, Svratka, Svitava and Trkmanka rivers), in the Czech Rebublic. Samples of water, sediment (Tab. 1.) and fish muscle were obtained during the seasons of 2012-2013. As the reference species were chosen chub (Squalius cephalus) and roach (Rutilus rutilus) due to their occurrence in all of the evaluated fishing grounds. The potential ecological risk analysis of heavy metal concentrations in the sediments indicated that nine sites in the middle and lower reaches posed moderate or considerable ecological risk. The health risk analysis of individual heavy metals in fish muscle indicated safe levels for the general population, but there was a possible risk in terms of the total target hazard quotients. The contents of the analysed metals in fish muscles were low at seven sites and did not exceed the values of limits admissible in the Czech Republic. The increased values of heavy metals in the fish muscle from the Svratka and Svitava rivers may be mainly derived from metal processing, electroplating industries, industrial wastewater and domestic sewage.


Defining the metabolic and growth responses of porcine haemophili to exogenous pyridine nucleotides and precursors.
A variety of biologically important pyridine nucleotides and precursors were examined for their capacities to satisfy the V-factor requirement of 30 strains of porcine haemophili. Of the compounds tested, only NAD, NMN and nicotinamide riboside (NR) supported the growth of all strains; NADP supported the growth of only the type strain of Haemophilus parasuis. Further studies with the H. parasuis type strain and the neotype strain of H. pleuropneumoniae demonstrated that, during growth, these organisms exhibited affinities for NMN that were greater than those for NAD; the affinity of H. pleuropneumoniae for NR was similar to that for NMN, whereas H. parasuis exhibited relatively low affinity for NR. With either organism, equimolar amounts of NAD and NMN supported the production of approximately equal amounts of biomass whereas growth yields were substantially lower when NR was the pyridine nucleotide source. When either organism was grown in the presence of excess exogenous [carbonyl-14C]NAD, cessation of growth was accompanied by the apparent exhaustion of the NAD supply. Approximately 80% of the radioactivity added as [14C]NAD could be recovered as extracellular [14C]nicotinamide and the majority of the assimilated radioactive material was present intracellularly in the form of a [14C]NAD(P) pool. The results are discussed in terms of the structural features required of a pyridine compound for it to support the growth of porcine haemophili, the capacity of these organisms to compete for pyridine nucleotide sources in vivo, and possible mechanisms involved in the assimilation of such compounds.


Cholecalciferol or Calcifediol in the Management of Vitamin D Deficiency.
Vitamin D deficiency is a global health problem due to its high prevalence and its negative consequences on musculoskeletal and extra-skeletal health. In our comparative review of the two exogenous vitamin D supplementation options most used in our care setting, we found that cholecalciferol has more scientific evidence with positive results than calcifediol in musculoskeletal diseases and that it is the form of vitamin D of choice in the most accepted and internationally recognized clinical guidelines on the management of osteoporosis. Cholecalciferol, unlike calcifediol, guarantees an exact dosage in IU (International Units) of vitamin D and has pharmacokinetic properties that allow either daily or even weekly, fortnightly, or monthly administration in its equivalent doses, which can facilitate adherence to treatment. Regardless of the pattern of administration, cholecalciferol may be more likely to achieve serum levels of 25(OH)D (25-hydroxy-vitamin D) of 30-50 ng/mL, an interval considered optimal for maximum benefit at the lowest risk. In summary, the form of vitamin D of choice for exogenous supplementation should be cholecalciferol, with calcifediol reserved for patients with liver failure or severe intestinal malabsorption syndromes.


A 3-D ultrasound-based framework to characterize the echo morphology of carotid plaques.
Carotid atherosclerosis is the main cause of brain stroke, which is the most common life-threatening neurological disease. Nearly all methods aiming at assessing the risk of plaque rupture are based on its characterization from 2-D ultrasound images, which depends on plaque geometry, degree of stenosis, and echo morphology (intensity and texture). The computation of these indicators is, however, usually affected by inaccuracy and subjectivity associated with data acquisition and operator-dependent image selection. To circumvent these limitations, a novel and simple method based on 3-D freehand ultrasound is proposed that does not require any expensive equipment except the common scanner. This method comprises the 3-D reconstruction of carotids and plaques to provide clinically meaningful parameters not available in 2-D ultrasound imaging, namely diagnostic views not usually accessible via conventional techniques and local 3-D characterization of plaque echo morphology. The labeling procedure, based on graph cuts, allows us to identify, locate, and quantify potentially vulnerable foci within the plaque. Validation of the characterization method was made with synthetic data. Results of plaque characterization with real data are encouraging and consistent with the results from conventional methods and after inspection of surgically removed plaques.


Regions of the Streptococcus sobrinus spaA gene encoding major determinants of antigen I.
Surface protein antigen A (SpaA), also called antigen B, antigen I/II, or antigen P1, is an abundant cell envelope protein that is the major antigenic determinant of Streptococcus sobrinus and other members of the Streptococcus mutans group of cariogenic bacteria. This laboratory has previously reported the cloning and expression in Escherichia coli of a BamHI restriction fragment of S. sobrinus DNA containing most of the spaA gene (pYA726) and encoding antigen I. Regions of spaA encoding immunodeterminants of antigen I were analyzed by either deletion mapping or expressing selected restriction fragments from the trc promoter. SpaA proteins produced by mutants harboring nested deletions, constructed by BAL 31 exonuclease treatment at a unique SstI site located towards the 3' end of the gene, were examined by Western immunoblot with rabbit serum against SpaA from S. sobrinus. Only SpaA polypeptides larger than 56 kilodaltons reacted with anti-SpaA serum. Various restriction fragments of the region of spaA encoding the antigenic determinants were cloned into an expression vector. The immunoreactive properties of the polypeptides encoded by those fragments indicated that expression of the immunodominant determinant required topographically assembled residues specified by noncontiguous regions located within 0.48-kilobase PvuII-to-SstI and 1.2-kilobase SstI-to-HindIII fragments which were adjacent on the spaA map.


A longitudinal view of factors that influence the emotional well-being of family caregivers to individuals with heart failure.
Caring for community-residing patients with heart failure can affect caregivers' emotional wellbeing. However, few studies have examined caregivers' well-being longitudinally, or identified factors associated with positive and negative outcomes. The objective of this longitudinal cohort study was to examine changes in caregivers' well-being over time, and to identify patient and caregiver factors associated with positive and negative outcomes. Fifty caregiver/heart failure patient dyads were recruited from an acute care facility and followed in the community. All participants completed surveys at hospital admission and 3, 6 and 12 months later. Caregivers completed assessments of depression symptoms and positive affect and standardized measures to capture assistance provided, mastery, personal gain, social support, participation restriction, and patients' behavioral and psychological symptoms. From patients, we collected demographic characteristics and health-related quality of life. Individual Growth Curve modelling was used to analyze the data. Caregivers' negative and positive emotions remained stable over time. Depression symptoms were associated with higher participation restriction in caregivers. Positive affect was associated with more personal gain and more social support. Patients' health-related quality of life and their behavioral and psychological symptoms were not significantly associated with caregivers' emotional outcomes. Interventions should be offered based on caregivers' needs rather than patients' health outcomes, and should focus on fostering caregivers' feelings of personal gain, assisting them with securing social support, and engaging in valued activities.


Immune response to influenza A(H1N1)v in HIV-infected patients.
HIV infection is considered a risk factor for severe outcomes of influenza A(H1N1)v infection. However, data on immune response against influenza A(H1N1)v virus in HIV-infected patients are lacking. Data from seven HIV-positive and 14 HIV-negative patients infected with A(H1N1)v and from 23 HIV-positive and six HIV-negative asymptomatic controls were analyzed to evaluate the clinical picture, A(H1N1)v viral shedding, and the immune response against the virus. Patients displayed mainly upper respiratory tract diseases (57.1%), while pneumonia was diagnosed only in HIV-negative patients (23.8% of subjects, of which 4.8% required intensive care unit admission). At day seven, 29% of HIV-infected patients were still positive for A(H1N1)v by RT-PCR on nasopharyngeal swabs. Interestingly, a persistence of CXCL10 secretion at high level and lower IL-6 levels was observed in HIV-positive subjects. The geometric mean haemagglutination inhibition titer (HI-GMT) and anti-influenza IgM levels were lower in HIV-positive individuals while anti-influenza IgG levels remained similar in the two groups. The immune impairment due to HIV infection could affect A(H1N1)v clearance and could lead to a lower antibody response and a persistent secretion of CXCL10 at high levels. However, the lower IL-6 secretion and treatment with highly active antiretroviral therapy (HAART) could result in a milder clinical picture.


Profile of drug interactions in hospitalized children.
The expected therapeutic response may be affected by the presence of drug interactions. With the high number of reports on new drug interactions, it has been difficult for health professionals to keep constantly updated. For this reason, computer systems have helped identify such interactions. To verify the rate and profile of drug interactions in medical prescriptions to hospitalized pediatric patients. A descriptive study investigated prescriptions to hospitalized pediatric patients. The study included patients between 0 and 12 years old, containing 4 or more drugs in their prescriptions. The analysis of interaction and incompatibility possibilities in prescribed drugs used Micromedex / Drug-Reax® program. From 2005 to 2006, 3,170 patients were investigated, and 11,181 prescriptions were analyzed, a mean value of 3.5 prescriptions/patient. In total, 6,857 drug interactions were found, which corresponds to 1.9 interaction/prescription. Among them, relevance to ampicillin and gentamicin, found in 220 (3.2%) prescriptions. In total, 2,411 drug incompatibilities in via y were found, a mean value of 0.5/prescription, with emphasis on vancomycin and cefepime, found in 243 (10.0%) prescriptions. The presence of drug interactions is a permanent risk in hospitals. This way, the utilization of computer programs, pharmacotherapy monitoring of patients and the pharmacist presence in the multidisciplinary team are some manners of contributing to hospitalized patients' treatment.


Dual Diagnosis of Trichohepatoenteric Syndrome and Lipoid Proteinosis in a Turkish Child.
Trichohepatoenteric syndrome (THES) is caused by pathogenic mutations in TTC37 and SKIV2L genes and characterized by intractable diarrhea, facial dysmorphism, hair abnormality, immunodeficiency, and skin abnormalities. Lipoid proteinosis is caused by pathogenic mutations in ECM1 gene and characterized by deposition of hyaline-like material in various tissues resulting in heterogenous clinical findings. Four years after the diagnosis and management of THES, due to new clinical findings, another reason for underlying features of the patient was considered. WES was performed and a homozygous c.507delT (p.Arg171GlyfsTer7) mutation in the ECM1 gene was detected. This case provides an example of co-existence of multiple genetic defects in a single patient born to consanguineous parents.


5HT7 receptors in the rodent suprachiasmatic nucleus.
Serotonergic modulation of circadian rhythms in rodent model preparations has received considerable attention over the past decade. Investigators have also been trying to determine which of the many serotonin receptor subtypes may be mediating the effects of serotonin in the suprachiasmatic nucleus, the location of the biological clock that generates the circadian rhythms. A single study in 1993 using the in vitro rat hypothalamic slice preparation suggested that serotonergic modulation of circadian rhythms at the level of the suprachiasmatic nucleus was acting via the newly discovered 5HT7 receptor subtype. Since that initial claim, serotonin modulation of circadian rhythms at the level of the suprachiasmatic nucleus has generally been attributed to 5HT7 receptor activation. However, when trying to cite relevant literature in support of 5HT7 involvement, it becomes evident that attributing rhythm-related serotonin activity in the suprachiasmatic nucleus to 5HT7 receptors may be somewhat premature. There are issues related to pharmacological specificity, species-specific results, and significant knowledge gaps that necessitate a careful review of the literature to make a judgment as to whether 5HT7 receptors are responsible for serotonergic activity in the rodent suprachiasmatic nucleus. In addition, there is sufficient data available at present to make an initial determination as to the degree of 5HT7 receptor involvement at any level in the generation or modulation of circadian rhythms in rodent species.


Bone morphogenetic protein-2 in biodegradable gelatin and β-tricalcium phosphate sponges enhances the in vivo bone-forming capability of bone marrow mesenchymal stem cells.
Bone marrow mesenchymal stem cells (MSCs) have been used for bone tissue engineering due to their osteogenic differentiation capability, but their application is controversial. To enhance their capability, we prepared biodegradable gelatin sponges incorporating β-tricalcium phosphate ceramics (GT sponge), which has been shown to possess excellent controlled drug-release properties. The GT sponge was used as a carrier for both rat MSCs and bone morphogenetic protein-2 (BMP-2) and osteogenic differentiation was assessed by subcutaneous implantation of four different kinds of implants, i.e. GT-alone, MSC-GT composites, BMP-GT composites and BMP-GT composites supplemented with MSCs (BMP-MSC-GT) in rats. Two weeks after implantation, histological sections showed new bone formation in the peripheral parts of the BMP-GT and in almost the total volume of the BMP-MSC-GT implants. After 4 weeks, histology as well as microCT analyses demonstrated extensive bone formation in BMP-MSC-GT implants. Gene expression and biochemical analyses of both alkaline phosphatase and bone-specific osteocalcin confirmed the histological findings. These results indicate that the combination of MSCs, GT and BMP synergistically enhances osteogenic capability and provides a rational basis for their clinical application in bone reconstruction.


Non-selective cation channels in basolateral-membrane vesicles from pars recta of rabbit kidney proximal tubule.
The characteristics of 86Rb+ fluxes through conductive channels in basolateral-membrane vesicles isolated from pars recta of rabbit kidney proximal tubule were investigated. In RbCl-, KCl- and NaCl-loaded vesicles a transient and almost equal accumulation of 86Rb+ was observed. The uptakes of 86Rb+ were inhibited to the same extent by 10 mM-BaCl2 in all loadings. The accumulation was driven by an electrical diffusion potential. The 86Rb+ flux was dependent on intravesicular Ca2+. Increasing concentrations of Ca2+ gradually decreased the 86Rb+ uptake. At 10 microM-Ca2+ the radioisotope flux was below 20% of control. The vesicles containing the channel showed very low selectivity among the univalent cations K+, Rb+, Li+, Na+ and choline+.


Pomegranate extract: a potential protector against aminoglycoside ototoxicity.
To investigate the effectiveness of pomegranate extract as protection against aminoglycoside ototoxicity. Prospective, randomised, controlled, experimental study. Eighteen Wistar albino rats were randomly allocated to 5 days of either: saline injections; gentamicin injections; or pomegranate extract (100 µl/day via gavage) plus gentamicin injections. Distortion product otoacoustic emissions were tested before treatment and on day 3. After treatment, reactive oxygen species levels were measured in each rat's right cochlea and right kidney via chemiluminescence. Baseline emission amplitudes were similar. Post-treatment emissions differed significantly in the two treatment groups (p < 0.001). Cochlear reactive oxygen species levels were significantly higher in the gentamicin group (mean ± standard deviation, 316.6 ± 36.5 relative light units per mg) than the gentamicin plus pomegranate extract group (240 ± 24.6 relative light units per mg) (p = 0.004); control group levels were 119.1 ± 10.3 relative light units per mg. Renal reactive oxygen species levels were similar for the control and gentamicin plus pomegranate extract groups (p = 0.59) but much higher in the gentamicin group (p = 0.004). Concurrent systemic pomegranate extract administration reduced reactive oxygen species level increases and otoacoustic emission changes, following aminoglycoside injection.


Adjuvant sequential chemotherapy with doxorubicin plus cyclophosphamide, methotrexate, and fluorouracil (ACMF) with concurrent radiotherapy in resectable advanced breast cancer.
Doxorubicin (Adriamycin) is an anthracycline effective in breast cancer. Despite a worldwide acceptance of Adriamycin in the adjuvant chemotherapy to maximize the survival benefit in the higher risk patients with breast cancer with promising results, oncologists in general do not favorably consider anthracyclines in the adjuvant treatment setting because of concern about the acute and chronic drug-related toxicity. For high-risk patients with breast cancer with more than three positive axillary lymph nodes, this series adopted a modified sequential regimen of ACMF first with Adriamycin (A) as a single agent in 3-weekly administration for three courses, and then a combination of cyclophosphamide, methotrexate, fluorouracil (CMF) every 3 to 4 weeks for six courses given in an outpatient setting concurrent with radiation therapy as an adjuvant treatment. A total of 56 patients underwent modified radical mastectomy and 3 others breast conservation surgery for their invasive breast cancer. Forty-seven (84%) patients completed the intended adjuvant treatment and 1 patient died of infection from treatment-related neutropenia. As a whole, the 3-year overall survival and disease-free survival rates of 56 patients analyzed were 82.3% and 64.4%, respectively. In this high-risk group, patients with four to nine positive nodes showed a slightly better trend of survival than those with 10 or more positive nodes without reaching statistically significant difference (36-month overall survival: 90.9% vs. 72.5%, p = 0.06; disease-free survival: 78.7% vs. 47.8%, p = 0.38). In this entire group of patients, locoregional recurrence was absent. A total of 55 episodes of grade III and IV hematologic toxicity were observed, with only one death from neutropenic sepsis. This modified ACMF regimen offers a good survival rate in breast cancer patients with more than three positive axillary lymph nodes. When these patients are carefully managed, the morbidity and mortality related to the treatment are low.


Implementing a self care program. Effect on employee health care utilization.
1. One way to reduce health care costs is to reduce the demand for health care services. This can be accomplished by teaching employees to make better decisions about when they should see the health care provider or go to the emergency department versus treating themselves at home using self care. 2. In an effort to reduce health care costs, a manufacturing company implemented a self care program using a publication called the HealthyLife Self Care Guide. The guide was distributed to employees during a 50 minute workshop. 3. Analysis of claims data 1 year prior to distribution of the Guide and 1 year after distribution showed a savings of $39.65 per employee (a 24.4% decrease in costs) due to reduced health care provider and emergency department visits. This amounted to a return on investment of 2.6:1. 4. It appears that implementing a self care program in a worksite setting can be an effective way to reduce employer health care costs.


Conversion of proinsulin to insulin occurs coordinately with acidification of maturing secretory vesicles.
Proinsulin is a single polypeptide chain composed of the B and A subunits of insulin joined by the C-peptide region. Proinsulin is converted to insulin during the maturation of secretory vesicles by the action of two proteases and conversion is inhibited by ionophores that disrupted intracellular H+ gradients. To determine if conversion of prohormone to hormone actually occurs in an acidic secretory vesicle, cultured rat islet cells were incubated in the presence of 3-(2,4-dinitroanilino)-3' amino-N-methyldipropylamine (DAMP), a basic congener of dinitrophenol that concentrates in acidic compartments and is retained there after aldehyde fixation. The cells were processed for indirect protein A-gold colocalization of DAMP, using a monoclonal antibody to dinitrophenol, and proinsulin, using a monoclonal antibody that exclusively reacts with the prohormone. The average density of DAMP-specific gold particles in immature secretory vesicles that contained proinsulin was 71/micron 2 (18 times cytoplasmic background), which indicated that this compartment was acidic. However, the density of DAMP-specific gold particles in the insulin-rich mature secretory vesicle averaged 433/micron 2. This suggests that although proinsulin conversion occurs in an acidic compartment, the secretory vesicles become more acidic as they mature. Since the concentration of anti-proinsulin IgG binding in secretory vesicles is inversely proportional to the conversion of proinsulin to insulin, we were able to determine that maturing secretory vesicles had to reach a critical pH before proinsulin conversion occurred.


The State of the LPO-AOD System and the Relative Length of Telomeric Repeats in the Chromosomes of Blood Leukocytes in Obstructive Sleep Apnea Syndrome.
We analyzed the relative length of telomeric repeats in peripheral blood leukocytes and indicators of oxidative stress in 32 men (mean age 51.2±3.1 years) with obstructive sleep apnea syndrome (OSAS). The control group consisted of volunteers without OSAS. The relative length of telomeres was determined in a DNA sample isolated from venous whole blood samples. The length of telomeric repeats of chromosomes was determined using quantitative real-time PCR (3 times for each DNA sample); albumin served as an internal control. The intensity of LPO processes and antioxidant protection was assessed by conventional spectrophotometric methods. It was found that in patients with OSAS, the relative length of telomeric repeats was lower by 48%, the levels of substrates and products of LPO were higher (double bonds and diene conjugates by 34 and 19%, respectively), and antioxidant protection indicators were lower (concentrations of fat-soluble vitamins α-tocopherol and retinol by 18 and 19.3%, respectively, total antioxidant activity by 32.9%) than in volunteers of the control group. Thus, we can conclude that the nucleotide sequences are reduced in patients with OSAS under conditions of intensified free radical oxidation.


Nefazodone in social phobia.
A variety of drug treatments have been shown to be effective in the treatment of social phobia. This study attempted to assess the efficacy of nefazodone, a new novel serotonergic drug, in the treatment of social phobia. Nefazodone was administered to 23 patients who had a primary DSM-IV diagnosis of social phobia, generalized type (diagnosed by the Structured Clinical Interview for DSM-IV), in a 12-week open clinical trial. Treatment began at 100 mg of nefazodone daily and was increased according to clinical response and side effects. Patients completed self-report measures at baseline and at weeks 4, 8, and 12. These measures included the Fear of Negative Evaluation scale, the Social Avoidance and Distress scale, the Social Anxiety Thoughts Questionnaire, the Fear Questionnaire, the State-Trait Anxiety Inventory, the Beck Depression Inventory, the Social Adjustment Scale Self-Report, and the Sheehan Disability Scale. Clinicians completed the Liebowitz Panic and Social Phobic Disorders rating form and the Brief Social Phobia Scale. Twenty-one of the 23 patients completed the 12-week trial. Sixteen (69.6%) were considered responders (moderate or marked improvement), and 7 (30.4%) were considered to be nonresponders (minimal improvement or no change in symptoms). Measures of social anxiety, social phobic avoidance, depression, and social functioning showed a statistically significant change at endpoint. These findings support a role for nefazodone in the treatment of social phobia, generalized type. Controlled studies will be required to further investigate this preliminary finding as well as to compare nefazodone with other pharmacologic treatments of social phobia.


An in vitro RNA editing system from cauliflower mitochondria: editing site recognition parameters can vary in different plant species.
Most of the 400 RNA editing sites in flowering plant mitochondria are found in mRNAs. Consequently, the sequence vicinities of homologous sites are highly conserved between different species and are presumably recognized by likewise conserved trans-factors. To investigate the evolutionary adaptation to sequence variation, we have now analyzed the recognition elements of an editing site with divergent upstream sequences in the two species pea and cauliflower. This variation is tolerated at the site selected, because the upstream cis-elements reach into the 5'-UTR of the mRNA. To compare cis-recognition features in pea and cauliflower mitochondria, we developed a new in vitro RNA editing system for cauliflower. In vitro editing assays with deleted and mutated template RNAs show that the major recognition elements for both species are located within the conserved sequence. In cauliflower, however, the essential upstream nucleotides extend further upstream than they do in pea. In-depth analysis of single-nucleotide mutations reveals critical spacing of the editing site and the specific recognition elements, and shows that the +1 nucleotide identity is important in cauliflower, but not in pea.


Photosystem II photoinactivation, repair, and protection in marine centric diatoms.
Diatoms are important contributors to aquatic primary production, and can dominate phytoplankton communities under variable light regimes. We grew two marine diatoms, the small Thalassiosira pseudonana and the large Coscinodiscus radiatus, across a range of temperatures and treated them with a light challenge to understand their exploitation of variable light environments. In the smaller T. pseudonana, photosystem II (PSII) photoinactivation outran the clearance of PSII protein subunits, particularly in cells grown at sub- or supraoptimal temperatures. In turn the absorption cross section serving PSII photochemistry was down-regulated in T. pseudonana through induction of a sustained phase of nonphotochemical quenching that relaxed only slowly over 30 min of subsequent low-light incubation. In contrast, in the larger diatom C. radiatus, PSII subunit turnover was sufficient to counteract a lower intrinsic susceptibility to photoinactivation, and C. radiatus thus did not need to induce sustained nonphotochemical quenching under the high-light treatment. T. pseudonana thus incurs an opportunity cost of sustained photosynthetic down-regulation after the end of an upward light shift, whereas the larger C. radiatus can maintain a balanced PSII repair cycle under comparable conditions.


The efficacy of pediatric blood culture sets in the determination of burn bacteremia.
A blood culture is an essential laboratory procedure necessary to confirm a septic episode. However, it is important to collect the blood sample at the appropriate time with an acceptable technique. The standard method is to collect at least 5 to 10 ml blood per culture bottle from patients with fevers. However, this volume of blood is an unrealistic amount to take from the frequently febrile pediatric patient. Alternatively, the pediatric blood culture bottle allows the collection of 1 ml blood per bottle to perform the same evaluation. We evaluated the two techniques of blood-culture collection over a 9-month period and compared the results between adult and pediatric blood culture bottles. Seventy-six patients, from November 1988 through February 1989, had blood cultures performed with the adult culture bottles, which produced a total of 1314 samples. A total of 113 patients, from March through July 1989, had blood cultures performed with the pediatric culture bottles, which produced a total of 758 samples. Percent recovery for the adult bottles versus the pediatric bottles was 13.95% versus 22.8% (p less than 0.0001). Since the amount of blood necessary to isolate an infectious agent is critical not only for laboratory identification but also for the volume of blood of pediatric patients, these data clearly establish the efficacy of pediatric blood culture bottles and the utilization of smaller amounts of blood. Not only did this approach significantly enhance organism recovery rate, but it may well be more cost-effective because fewer cultures need to be performed to isolate the infectious organism.