Early results of selective laser trabeculoplasty in patients resistant to deep sclerectomy.
In this study, we aimed to determine the early results of selective laser trabeculoplasty to trabeculo-Descemet membrane in patients resistant to deep sclerectomy (DS) surgery. Twelve eyes of 12 patients that had undergone DS for primary open-angle glaucoma were enrolled. Preoperative and postoperative intraocular pressures (IOP) were measured by Goldmann applanation tonometer. Time to failure after DS was noted. Failure is described as IOP higher than 20 mm Hg. All patients underwent selective laser trabeculoplasty (SLT) after failure. Eight to 10 applications of SLT were applied over the trabeculo-Descemet membrane. Laser was applied to the surgical site only. The IOPs before SLT, at postoperative first week, first month, and third month were measured. Mean follow-up time was 23.5 ± 9.5 months. Mean preoperative IOP was 34.2 ± 6.7 mm Hg; mean postoperative IOP at first week was 12.7 ± 3.2 mm Hg. Mean time to failure after DS procedure was 21.6 ± 7.6 months. Mean IOP was 20.6 ± 1.9 mm Hg just before SLT procedure. Mean IOP measurements after SLT at first week, first month, and third month were 10.5 ± 2.2 mm Hg, 10.5 ± 1.9 mm Hg, and 10.9 ± 2.1 mm Hg, respectively. Selective laser trabeculoplasty after DS is an effective and noninvasive procedure that can be done on an outpatient basis. Our results suggest that SLT applied to the surgical site is an effective intervention in the short term but long-term results are required to assess the efficacy.


Real-Time Internet of Medical Things System for Detecting Blood Leakage during Hemodialysis Using a Novel Multiple Concentric Ring Sensor.
Venous needle dislodgement (VND) is a major healthcare safety concern in patients undergoing hemodialysis. Although VND is uncommon, it can be life-threatening. The main objective of this study was to implement a real-time multi-bed monitoring system for VND by combining a novel leakage-detection device and IoMT (Internet of Medical Things) technology. The core of the system, the Acusense IoMT platform, consisted of a novel leakage-detection patch comprised of multiple concentric rings to detect blood leakage and quantify the leaked volume. The performance of the leakage-detection system was evaluated on a prosthetic arm and clinical study. Patients with a high risk of blood leakage were recruited as candidates. The system was set up in a hospital, and the subjects were monitored for 2 months. During the pre-clinical simulation experiment, the system could detect blood leakage volumes from 0.3 to 0.9 mL. During the test of the IoMT system, the overall success rate of tests was 100%, with no lost data packets. A total of 701 dialysis sessions were analyzed, and the accuracy and sensitivity were 99.7% and 90.9%, respectively. Evaluation questionnaires showed that the use of the system after training changed attitudes and reduced worry of the nursing staff. Our results show the feasibility of using a novel detector combined with an IoMT system to automatically monitor multi-bed blood leakage. The innovative concentric-circle design could more precisely control the warning blood-leakage threshold in any direction to achieve clinical cost-effectiveness. The system reduced the load on medical staff and improved patient safety. In the future, it could also be applied to home hemodialysis for telemedicine during the era of COVID-19.


Ventricular pacing via a Swan Ganz catheter: a new mode of pacemaker therapy.
Hemodynamic monitoring via a Swan Ganz catheter is often required with critically-ill patients. In the further course of the illness emergency temporary pacing might be necessary, most frequently for AV-conduction disturbances or sinus bradycardia. Insertion via the right internal jugular vein has been used in many cases. This raises the problem that one may find a second central vein, which can lead to serious problems and in any case may be a dangerous waste of time. Recently we had the opportunity to stimulate the right ventricle with a small pacing wire, brought into the ventricle via a Swan Ganz catheter especially designed for this purpose with an extra lumen ending in the right ventricle of the heart. In this account we describe our experience with the first ten patients to whom this method was applied.


How Cues of Being Watched Promote Risk Seeking in Fund Investment in Older Adults.
Social cues, such as being watched, can subtly alter fund investment choices. This study aimed to investigate how cues of being watched influence decision-making, attention allocation, and risk tendencies. Using decision scenarios adopted from the "Asian Disease Problem," we examined participants' risk tendency in a financial scenario when they were watched. A total of 63 older and 66 younger adults participated. Eye tracking was used to reveal the decision-maker's attention allocation (fixations and dwell time per word). The results found that both younger and older adults tend to seek risk in the loss frame than in the gain frame (i.e., framing effect). Watching eyes tended to escalate reckless gambling behaviors among older adults, which led them to maintain their share in the depressed fund market, regardless of whether the options were gain or loss framed. The eye-tracking results revealed that older adults gave less attention to the sure option in the eye condition (i.e., fewer fixations and shorter dwell time). However, their attention was maintained on the gamble options. In comparison, images of "watching eyes" did not influence the risk seeking of younger adults but decreased their framing effect. Being watched can affect financial risk preference in decision-making. The exploration of the contextual sensitivity of being watched provides us with insight into developing decision aids to promote rational financial decision-making, such as human-robot interactions. Future research on age differences still requires further replication.


4D printing: a cutting-edge platform for biomedical applications.
Nature's materials have evolved over time to be able to respond to environmental stimuli by generating complex structures that can change their functions in response to distance, time, and direction of stimuli. A number of technical efforts are currently being made to improve printing resolution, shape fidelity, and printing speed to mimic the structural design of natural materials with three-dimensional printing. Unfortunately, this technology is limited by the fact that printed objects are static and cannot be reshaped dynamically in response to stimuli. In recent years, several smart materials have been developed that can undergo dynamic morphing in response to a stimulus, thus resolving this issue. Four-dimensional (4D) printing refers to a manufacturing process involving additive manufacturing, smart materials, and specific geometries. It has become an essential technology for biomedical engineering and has the potential to create a wide range of useful biomedical products. This paper will discuss the concept of 4D bioprinting and the recent developments in smart materials, which can be actuated by different stimuli and be exploited to develop biomimetic materials and structures, with significant implications for pharmaceutics and biomedical research, as well as prospects for the future.


Ethnic and gender differences in risk factors for smoking onset.
The purpose of this investigation was to explore gender and ethnic differences in the prevalence of risk factors for smoking onset. As part of a larger study, 6,967 seventh graders completed a questionnaire measuring variables shown in previous research to predict cigarette smoking. The results indicated that at this age level, risk factors for smoking initiation are less prevalent among African Americans than European American youth and among girls relative to boys. These findings are consistent with the lower incidence of smoking among African Americans and girls at this young age. Future research should examine changes in risk factors over time to determine whether such changes accompany the rising smoking rate typically observed among girls and African Americans later in adolescence.


Design of a pilot study to evaluate tandem mass spectrometry for neonatal screening.
In recent years, tandem mass spectrometry has generated great interest as a method for neonatal screening. The basic principle is electronically controlled separation of analytes by their mass-to-charge ratio. The advantage of this detection system is speed, the capability to analyze for many different compounds in a single analysis, and a minimal need for auxilliary assay reagents. The prevailing screening technique uses stable isotope dilution, butylesterification, and MS/MS analysis to quantify amino acids and acylcamitines in neonatal dried blood spot samples. This allows detection of more than 30 inborn errors of metabolism of amino acids, fatty acids, and other organic acids. In Denmark, a large-scale pilot study is being implemented to evaluate the screening potential of tandem mass spectrometry. National patient registers and medical records from clinical genetics units are used to identify cohorts of healthy infants and infants with selected inborn errors of metabolism. The neonatal screening samples of these infants are retrieved from a biobank and are assayed for amino acids and acylcarnitines using tandem mass spectrometry. This study yields decision values for neonatal screening, which will be evaluated in a subsequent 2-year prospective pilot study, offering the test to 140,000 neonates as a voluntary adjunct to the existing screening program. The organization consists of integrated units for neonatal screening and clinical genetics. The effect of the program will be assessed in terms of screening efficiency, cost and short term clinical outcome.


[The assay of enzyme activity in EIA--colorimetric, fluorometric and luminometric method].
The method of enzyme activity related in enzyme immunoassay was described. The detection methods were colorimetric, fluorometric and chemi- or bioluminometric assay for enzyme, and the sensitivity of three detection methods were compared. The order of detection limit was chemi- or bioluminometry > fluorometry > colorimetry. But it is important that the selection of detection method of enzyme will make fit on your desirable sensitivity of analysis.


Analgesia in adult trauma patients in physician-staffed Austrian helicopter rescue: a 12-year registry analysis.
Sufficient analgesia is an obligation, but oligoanalgesia (NRS> 3) is frequently observed prehospitally. Potent analgesics may cause severe adverse events. Thus, analgesia in the helicopter emergency medical service (HEMS) setting is challenging. Adequacy, efficacy and administration safety of potent analgesics pertaining to injured patients in HEMS were analysed. Observational study evaluating data from 14 year-round physician-staffed helicopter bases in Austria in a 12-year timeframe. Overall, 47,985 (34.3%) patients received analgesics, 26,059 of whom were adult patients, injured and not mechanically ventilated on site. Main drugs administered were opioids (n=20,051; 76.9%), esketamine (n=9082; 34.9%), metamizole (n=798; 3.1%) and NSAIDs (n=483; 1.9%). Monotherapy with opioids or esketamine was the most common regimen (n=21,743; 83.4%), while opioids together with esketamine (n= 3591; 13.8%) or metamizole (n=369; 1.4%) were the most common combinations. Females received opioids less frequently than did males (n=6038; 74.5% vs. n=14,013; 78.1%; p< 0.001). Pain relief was often sufficient (> 95%), but females more often had moderate to severe pain on arrival in hospital (n=34; 5.0% vs. n=59; 3.2%; p=0.043). Administration of potent analgesics was safe, as indicated by MEES, SpO2 and respiratory rates. On 10% of all missions, clinical patient assessment was deemed sufficient by HEMS physicians and monitoring was spared. Opioids and esketamine alone or in combination were the analgesics of choice in physician-staffed HEMS in Austria. Analgesia was often sufficient, but females more than males suffered from oligoanalgesia on hospital arrival. Administration safety was high, justifying liberal use of potent analgesics in physician-staffed HEMS.


Argatroban in short-term percutaneous ventricular assist subsequent to heparin-induced thrombocytopenia.
Heparin-induced thrombocytopenia paradoxically is a transient pro-thrombotic disorder triggered by heparin exposure. If not treated appropriately, it can be life threatening because of its related thromboembolic complications. In particular, it presents a unique challenge in patients needing extracorporeal life support, because anticoagulation is essential for safe management. This case report describes the safe, efficacious use of Argatroban during short-term support of a patient with a percutaneously inserted left ventricular assist TandemHeart device.


Sustainable Biocatalytic Procedure for Obtaining New Branched Acid Esters.
Biocatalytic synthesis of 2-ethylhexyl 2-methylhexanoate is described in this work for the first time. This branched-chain ester is suitable for use at low temperatures in numerous applications. The immobilized lipase Novozym® 435 has demonstrated its ability to catalyze the ester synthesis from 2-ethylhexanol and 2-methylhexanoic acid in a solvent-free medium. The high reaction times that are required result in a loss of alcohol by evaporation, which must be compensated for with an excess of this substrate if high conversions are to be achieved. Therefore, two strategies are established: 70 °C with a 10% excess of alcohol, which requires a longer operating time and provides conversions of 97%, and 80 °C with a 20% excess of alcohol, which allows for the achievement of a 99% conversion in a shorter time. The optimal reaction conditions have been chosen based on reusability of the enzyme, process productivity, green metrics and preliminary economic study. When the synthesis is carried out under the best conditions (70 °C, 10% molar excess of alcohol and six uses of the immobilized enzyme) a productivity of 203.84 kg product × kg biocatalyst-1 is attained. The biocatalytic procedure matches many of the objectives of "green chemistry" and is suitable to be scaled up and used in industrial manufacturing.


Stray radiation dose reduction using a backscatter shield and a rectangular collimator for a handheld intraoral X-ray unit.
The objective of this study was to study the dose reduction effect on occupational exposure of operators and public exposure other than operators when a backscatter shield and a rectangular collimator are used in conjunction with a handheld intraoral X-ray unit. The occupational exposure was reduced to 40% when the backscatter shield was attached to the cone-tip, to 13% when the rectangular collimator was attached and to 7.7% when the backscatter shield and rectangular collimator were used together. On the other hand, the public exposure was reduced to 20% when the rectangular collimator was attached, but the backscatter shield was not effective in reducing the public exposure. Attaching a backscatter shield is effective in reducing the occupational exposure, and a rectangular aperture is effective in reducing the occupational exposure, as well as the public exposure.


Breast-milk sodium as a predictor of breastfeeding patterns.
This study partially replicates and extends a study reporting that elevated breast-milk sodium BM [Na+] during early lactogenesis was predictive of poor breastfeeding outcomes. The present study used 6-day postpartum breast milk. Consistent with the findings of the earlier study, 80% of those with a BM [Na+] of 16 mmol/L or lower at day 6 sustained a high level of breastfeeding at week 4, compared to only 50% of those with an elevated BM [Na+] (chi 2 = 4.05, df = 1, p = .04). This difference was even greater in a subgroup of mothers predicted to be at high risk for insufficient milk supply on the basis of support density and self-perception variables. Of the latter group, 75% with low BM [Na+] sustained a high level of breastfeeding at 4 weeks postpartum, compared to only 22% with an elevated BM [Na+] (chi 2 = .65, df = 1, p = .01). In contrast, among the low-risk mothers BM [Na+] levels were not associated with any difference in breast-milk sustainment (89% and 82% sustainment for low- and high-sodium groups, respectively). Thus a normal drop in BM [Na+] is predictive of higher sustainment of breastfeeding. However, the predictive validity of this marker appears to be enhanced by combining it with the psychosocial variables of support density and self-perception of breastfeeding by the mother.


Stimulation of erythroid progenitors by high concentrations of erythropoietin results in normoblasts arrested in G2 phase of the cell cycle.
The cell cycle status of human erythroid precursors generated in a two-step liquid culture was studied by a double-labeling flow cytometric technique. Following a first phase, where peripheral blood mononuclear cells were cultured in the presence of a combination of growth factors, not including erythropoietin (Epo), the cells were washed and recultured in a second phase in the presence of Epo. This procedure resulted in a stimulation of the proliferation and maturation of erythroid precursors. In the presence of optimal concentrations of Epo (2 U/mL), a high percentage (> 40%) of cells were found in the S phase of the cell cycle until day 10. Then, as a result of maturation, the proportion of cells in S gradually decreased, reaching less than 2.0% by day 21. At this time, the culture consisted of > 95% hemoglobin-containing, nonproliferating, orthochromatic normoblasts. Cell cycle analysis of this normoblast population demonstrated a bimodal distribution; while the majority of the cells had a diploid (2C) DNA content, i.e., cells in G1 (or G0) phase, a sizable fraction was tetraploid (4C) corresponding to cells in G2. In contrast, in cultures stimulated with physiological concentrations of Epo (around 50 mU/mL), all the terminally differentiated cells were arrested at the G1 phase. These results suggest that Epo is an essential growth-promoting factor for erythroid precursors, but supraphysiological concentrations, such as present in vivo in severe anemia (e.g., aplastic anemia) or after Epo administration, may be associated with development of normoblasts with abnormal DNA content.


Antifungal properties of the immunosuppressant FK-506: identification of an FK-506-responsive yeast gene distinct from FKB1.
FK-506 is a novel and potent antagonist of T-cell activation and an inhibitor of fungal growth. Its immunosuppressive activity can be antagonized by the structurally related antibiotic rapamycin, and both compounds interact with cytoplasmic FK-506-binding proteins (FKBPs) in T cells and yeast cells. In this paper, we show that FK-506 and two analogs inhibit vegetative growth of Saccharomyces cerevisiae in a fashion that parallels the immunosuppressive activity of these compounds. Yeast mutants resistant to FK-506 were isolated, and at least three complementation groups (fkr1, fkr2, and fkr3) were defined. These fkr mutants show no alteration in their levels of FK-506-binding activity. Likewise, strains carrying null alleles of FKB1 (the yeast gene coding for the FKBP) remain FK-506 sensitive, indicating that depletion of yeast FKBP is not sufficient to confer an FK-506 resistance phenotype, although fkb1 null mutants are resistant to rapamycin. FKB1 does not map to the three fkr loci defined here. These results suggest that yeast FKBP mediates the inhibitory effect of rapamycin but that at least one other protein is directly involved in mediating the activity of FK-506. Interestingly, the ability of FK-506 to rescue a temperature-sensitive growth defect of the fkr3 mutant suggests that the FKR3 gene may define such a protein.


Diaphragmatic angiogenic growth factor mRNA responses to increased ventilation caused by hypoxia and hypercapnia.
This study investigates the effect of increased ventilation on the expression of messenger ribonucleic acid (mRNA) levels of vascular endothelial growth factor (VEGF), basic fibroblast growth factor (bFGF) and transforming growth factor-beta1 (TGF-beta1) in the diaphragm of intact, awake, spontaneously breathing rats, compared with responses in paralysed, mechanically-ventilated animals at similar blood gas and ventilatory levels. Four groups of intact, rats were studied in a body box, each group breathing one of four gases: room air, 12% oxygen (O2), 5% carbon dioxide (CO2), or 12% O2+5% CO2 for 1 h. Another 4 groups of paralysed, mechanically-ventilated animals were matched for arterial blood gas and ventilatory level. The results showed that VEGF mRNA abundance was increased three-fold and that of bFGF 1.5-fold when 12% O2+5% CO2 were breathed, but TGF-beta1 did not change. A significant linear relationship of VEGF and bFGF mRNA to minute ventilation was observed in awake animals (r=0.98, p<0.02 and r=0.87, p<0.03, respectively). The paralysed, mechanically-ventilated animals showed no mRNA increases for any probe. Systemic hypoxia had no additional effect on VEGF or bFGF levels in the diaphragm. It was concluded that messenger ribonucleic acid for vascular endothelial growth factor and basic fibroblast growth factor in the diaphragm rises significantly as a result of active ventilation and not due to blood gas/pH changes or to passive muscle shortening per se.


[Troponin and other markers of myocardial ischemia injury, what is the relevance in internal medicine?].
Troponin is now the gold standard for the diagnosis of myocardial infarction. Aiming at improving the management of a patient suspect of an acute coronary syndrome, this article will point the interpretation of troponin dosages according to the clinical presentation and concomitant diseases. First, the interest of troponin dosage as compared with other markers of myocardial ischemia will be underlined. Then, the literature available about troponin in cardiovascular diseases but also in extracardiac diseases will be analysed. Finally, the difficulties of assay will be discussed. The availability of a sensitive and specific marker such as troponin is definitively a progress in the management of patients with an acute coronary syndromes. But it remains a biological contribution to the global management of the patient. It is important to know the causes susceptible to increase the levels of troponin to avoid a wrong interpretation of the dosage, leading to diagnostic but also therapeutic mistakes.


R. v. Salford Health Authority Ex Parte Janaway, 1 December 1988.
The House of Lords of the UK dismissed the appeal of a medical secretary against a lower court decision (see Annual Review of Population Law, Volume 14, 1987, Section 240) that she could be terminated for refusing to type a letter referring a patient to a consultation for a possible abortion. It held that her refusal could not be justified under Section 4(1) of the Abortion Act 1967, which provides that no person may be forced to "participate in any treatment authorized by this Act to which he has a conscientious objection," because typing a letter does not constitute participation in treatment.


Evidence of unprecedented rise in growth synchrony from global tree ring records.
Changes in the temporal coherence between populations, which can influence their stability, resilience and persistence, remain a critical uncertainty of climate change. Recent studies have documented increasing spatial synchrony between populations at continental scales and linked it to anthropogenic climate change. However, the lack of long-term and global baseline perspectives on spatial synchrony presents a challenge to understanding the importance of these trends. Here, we show a steady rise in the spatial synchrony of annual tree growth from a global tree ring database over the past 50 years that is consistent across continents, species and environmental conditions and is unprecedented for the past millennium. Increasing growth synchrony coincided with warming trends and potentially rising synchrony in the temperature records. We discuss the potential driving mechanisms and the limitations in the interpretation of this trend, and we propose that increasing mutual dependency on external factors (also known as Moran's effect) linked to rising global temperatures is the most likely driver of more homogeneous global growth dynamics.


Phylogeography of pipistrelle-like bats within the Canary Islands, based on mtDNA sequences.
Evolution of three Canary Island Vespertilionid bat species, Pipistrellus kuhlii, Pipistrellus maderensis, and Hypsugo savii was studied by comparison of approximately 1 kbp of mtDNA (from cytochrome b and 16S rRNA genes) between islands. mtDNA reveals that both P. kuhlii and P. maderensis exist in sympatry on Tenerife (and possibly other islands). Their morphological similarity explains why their co-occurrence had not been detected previously. Levels of sequence divergence are quite low within P. maderensis. Haplotypes were either identical or separated by </=2 mutational steps on two of the islands (La Gomera and El Hierro). However there is sufficient between-island divergence between haplotypes from Tenerife, La Palma, and La Gomera/El Hierro to suggest that they could represent three evolutionary significant units (ESU). H. savii has an overlapping island distribution but a contrasting phylogeographical pattern. Most significantly, sequence divergence is greatest between La Gomera and El Hierro (>/=12 mutational steps) indicating colonization of the latter from the former sometime during the last approximately 1.2 Ma, with low subsequent gene flow. Unlike P. maderensis the El Hierro population alone appears to represent an ESU. The H. savii haplotypes detected in Gran Canaria and Tenerife are identical or separated by 1 mutational step.


Tender point scores and their relations to signs of mobility, symptoms, and disability in female home care personnel and the prevalence of fibromyalgia syndrome.
In this study of female home care personnel employed in a municipality (n = 643; participation rate 94%) we investigated (1) the prevalence of tender points and fibromyalgia (FM); (2) the relationships between tender point score and other signs and symptoms; (3) if subgroups based on the tender point score differed with respect to signs, symptoms, disability, and health related quality of life; and (4) signs that showed the strongest intercorrelations with disability and health. The following variables were registered: (1) Signs: joint mobility, spinal posture and mobility, tender points, and segmental mobility and pain provocation at L4-S1 levels of the low back. (2) SYMPTOMS: pain and pain intensity and other symptoms. (3) Disability (i.e., self-rated reduced capacity for everyday activities and employment) and health: 3 indices and sick leave. The tender point score correlated with the number of pain regions and the pain intensities, and the amount of other symptoms, sick leave, and disability. Tender point score was the strongest regressor of the investigated signs in regression of the 2 disability indices. Segmental pain showed the strongest correlation with tender point score. Three subgroups identified by tender point score showed significant differences in segmental pain, prevalence and intensity of different symptoms, disability, and health related quality of life. The prevalence of FM was 2.0%. Tender point score together with different symptoms showed relatively strong correlations with disability. A relatively high prevalence of FM was found in occupationally active female home care personnel.


A Four-dimensional Motion Field Atlas of the Tongue from Tagged and Cine Magnetic Resonance Imaging.
Representation of human tongue motion using three-dimensional vector fields over time can be used to better understand tongue function during speech, swallowing, and other lingual behaviors. To characterize the inter-subject variability of the tongue's shape and motion of a population carrying out one of these functions it is desirable to build a statistical model of the four-dimensional (4D) tongue. In this paper, we propose a method to construct a spatio-temporal atlas of tongue motion using magnetic resonance (MR) images acquired from fourteen healthy human subjects. First, cine MR images revealing the anatomical features of the tongue are used to construct a 4D intensity image atlas. Second, tagged MR images acquired to capture internal motion are used to compute a dense motion field at each time frame using a phase-based motion tracking method. Third, motion fields from each subject are pulled back to the cine atlas space using the deformation fields computed during the cine atlas construction. Finally, a spatio-temporal motion field atlas is created to show a sequence of mean motion fields and their inter-subject variation. The quality of the atlas was evaluated by deforming cine images in the atlas space. Comparison between deformed and original cine images showed high correspondence. The proposed method provides a quantitative representation to observe the commonality and variability of the tongue motion field for the first time, and shows potential in evaluation of common properties such as strains and other tensors based on motion fields.


In vitro reconstitution and analysis of the 6-deoxyerythronolide B synthase.
Notwithstanding an extensive literature on assembly line polyketide synthases such as the 6-deoxyerythronolide B synthase (DEBS), a complete naturally occurring synthase has never been reconstituted in vitro from purified protein components. Here, we describe the fully reconstituted DEBS and quantitatively characterize some of the properties of the assembled system that have never been explored previously. The maximum turnover rate of the complete hexamodular system is 1.1 min(-1), comparable to the turnover rate of a truncated trimodular derivative (2.5 min(-1)) but slower than that of a bimodular derivative (21 min(-1)). In the presence of similar concentrations of methylmalonyl- and ethylmalonyl-CoA substrates, DEBS synthesizes multiple regiospecifically modified analogues, one of which we have analyzed in detail. Our studies lay the foundation for biochemically interrogating and rationally engineering polyketide assembly lines in an unprecedented manner.


The distribution and kinetics of polyomavirus in lungs of intranasally infected newborn mice.
The primary cell types that sustain polyomavirus (Py) replication following intranasal infection as well as the nature of the host cellular response to Py were unknown. As this is an essential and specific site for virus entry, it seems likely that viral gene function must be adapted to these mucosal tissues. Using immunohistochemistry and in situ hybridization, we determined the cell types in the lung that support Py gene expression and replication following intranasal inoculation of newborn mice within 24 h of birth. Lungs were collected daily from days 1 to 10 postinfection for Py DNA and early T antigen analysis and for histological examination by H&E staining, using methods that preserve the delicate newborn lung architecture. Viral DNA was present in increasing quantities from 2 to 6 dpi in a subset of the Clara cells lining the inner lumen of the bronchi and bronchioles, while T antigen expression was present in a majority of the cells in the bronchi and bronchiole lumen. A distinct and transient pattern of hyperplasia was observed among the cells expressing T antigen and was present from 3 through 6 dpi. Py DNA-containing cells exfoliated into the bronchiole lumen and alveolar ducts, but Py T antigen was not detected in these cells. Py DNA was first detected at 2 dpi, increased through 6 dpi, and abruptly declined through 9 dpi at which time there was no sign of viral DNA in the lungs by in situ hybridization. An unusual infiltration of neutrophils began before the presence of exfoliated cells or Py replication and continued for 2-3 days and was followed by a lymphocytic infiltration at 8-10 dpi lasting 2-3 days. Neither the hyperplasia nor the neutrophil infiltration occurred following infection with the MOP1033 MT-Ag or RB1 LT-Ag mutants of Py. In addition, both the neutrophil infiltration and the transient hyperplasia are in stark contrast to the heavy macrophage infiltration that follows infection of lungs with mouse adenovirus. Thus it appears that Py elicits a distinct host response pattern not seen with other DNA viral infections.


Children's understanding of the relation between delayed video representation and current reality: a test for self-awareness?
This study investigated whether children's ability to recognize themselves in delayed video feedback indicates changes in self-awareness (Povinelli, Landau, & Perilloux, 1996, Child Development, 67, 1540-1554). Children were presented with 3-min-old videos of themselves to test whether they would investigate the current state of affairs upon seeing a surprising element in the video. In one condition, a sticker had been covertly placed into the child's hair, and in another an object had been hidden in a box. Both conditions proved equally difficult and performance correlated. Four-year-olds performed better than 3-year-olds, and children who failed the tasks retrieved the "surprise" item when presented with a mirror. There was no evidence to suggest that children's difficulties were due to immature metarepresentational thinking, lack of experience, problems with the questions, or problems appreciating the correspondence between image and referent. Yet, the parallel results in both conditions and the likelihood of false positives and false negatives indicate that the video test in its present form may not be a valid measure of differences in self-awareness.


Vergence position of rest: spontaneous variability.
We determined the vergence position of rest (horizontal component) in two sessions, 1 to 6 weeks apart. Each session contained 8 trials. Twenty observers with normal eyes looked through Risley prisms that allowed a continuous modulation of their strength. We asked the observers to look at a fully fusionable picture, and to adjust the prisms such that viewing appeared most relaxing. The viewing distances were 400 cm and 50 cm. The standard deviation among the 8 trials of each session, averaged over the two sessions, the viewing distances of 400 cm and 50 cm, and the 20 observers, was + or -1.4 prism dioptres (cm/m). From the first session to the second, many observers changed the preferred prism significantly. At a viewing distance of 400 cm, 10 of 20 observers changed their preferred prism by more than 1.0 prism diopter, up to 3.3 prism diopters. At a viewing distance of 50 cm, 11 of 20 observers changed their preferred prism by more than 1.0 prism diopter, up to 5.1 prism diopters. The prismatic strength estimated as being most relaxing can be very variable. Before considering a prescription, the practitioner should determine whether the preferred prismatic strength remains reasonably stable. To ascertain this consistency, more than one session seems to be necessary.


An integrated approach to methicillin-resistant Staphylococcus aureus control in a rural, regional-referral healthcare setting.
To curtail the prevalence and cross-transmission of methicillin-resistant Staphylococcus aureus (MRSA) in a rural healthcare setting. Before-after, quasi-experimental quality improvement study. A regional-referral hospital, 5 affiliated nursing homes, and an outpatient MRSA clinic. Residents of the 5 nursing homes were screened for MRSA at baseline and 1 year later. Active surveillance cultures were performed on subsequently admitted nursing home residents, "high-risk" patients admitted to the hospital, and household contacts of clinic patients. The decolonization regimen consisted of systemic therapy with minocycline and rifampin and topical therapy with nasal mupirocin ointment and 5% tea tree oil body wash. Three separate samples for cultures to document clearance of MRSA colonization were obtained at 1-week intervals 1 month after the completion of decolonization therapy. Samples for follow-up cultures were obtained at month 6 and month 12 after the completion of decolonization therapy. After intervention and follow-up for 12 months or more, the prevalence of MRSA carriage at the nursing homes decreased by 67% (P < .001), and 120 (82%) of 147 nursing home residents and 111 (89%) of 125 clinic patients remained culture-negative for MRSA. Twenty-three (24%) of 95 new clinic patients had at least 1 MRSA-positive contact. Mupirocin resistance did not develop. In the hospital, the incidence rate of nosocomial MRSA infection decreased from 0.64 infections per 1,000 patient-days before the interventions to 0.40 infections per 1,000 patient-days 1 year after the interventions and to 0.32 infections per 1,000 patient-days 2 years after the intervention (P < .01). Use of active surveillance cultures and decolonization therapy was effective in decreasing the prevalence of asymptomatic carriage, the incidence of nosocomial infection, and the overall prevalence of MRSA in our rural healthcare setting.


5-Oxoprolinal: transition-state aldehyde inhibitor of pyroglutamyl-peptide hydrolase.
Pyroglutamyl-peptide hydrolase (EC 188.8.131.52) removes the N-terminal pyroglutamyl residue from pyroglutamyl-containing peptides such as thyrotropin-releasing hormone (TRH), luteinizing hormone-releasing hormone (LH-RH), neurotensin, and bombesin. The aldehyde analogue of pyroglutamate, 5-oxoprolinal, was synthesized as an active site directed transition-state inhibitor of the enzyme. 5-Oxoprolinal was found to be a potent (Ki = 26 nM) and specific competitive inhibitor of pyroglutamyl-peptide hydrolase. Other aldehydes tested inhibited the enzyme only weakly or not at all. 5-Oxoprolinal blocked the degradation of LH-RH by purified pyroglutamyl-peptide hydrolase. The inhibitor, when injected into mice, inhibited the enzyme after 10 and 30 min. 5-Oxoprolinal should be of value in studies probing the biological significance of pyroglutamyl-peptide hydrolase.


Catheter-manometer system damped blood pressures detected by neural nets.
Degraded catheter-manometer systems cause distortion of blood pressure waveforms, often leading to erroneously resonant or damped waveforms, requiring waveforms quality control. We have tried multilayer perceptron back-propagation trained neural nets of varying architecture to detect damping on sets of normal and artificially damped brachial arterial pressure waves. A second-order digital simulation of a catheter-manometer system is used to cause waveform distortion. Each beat in the waveforms is represented by an 11 parameter input vector. From a group of normotensive or (borderline) hypertensive subjects, pressure waves are used to statistically test and train the neural nets. For each patient and category 5-10 waves are available. The best neural nets correctly classify about 75-85% of the individual beats as either adequate or damped. Using a single majority vote classification per subject per damped or adequate situation, the best neural nets correctly classify at least 16 of the 18 situations in nine test subjects (binomial P = 0.001). More importantly, these neural nets can always detect damping before clinically relevant parameters such as systolic pressure and computed stroke volume are reduced by more than 2%. Neural nets seem remarkably well adapted to solving such subtle problems as detecting a slight damping of arterial pressure waves before it affects waveforms to a clinically relevant degree.


Phytochrome Regulation of the Response to Exogenous Gibberellins by Epicotyls of Vigna sinensis.
The elongation rate of cowpea epicotyls from whole cowpea (Vigna sinensis) seedlings and derooted and debladed plants (explants) increased after the main light period (8-hour duration) was extended with either continuous low intensity tungsten light or brief (5 minutes) far-red (FR) irradiation. This end-of-day FR effect was reversed by red (R) irradiation suggesting the involvement of phytochrome. These results confirm and extend those obtained previously with other species. Localization studies indicate the epicotyl to be the site of the photoreceptor. Treatment of cowpea seedlings with paclobutrazol, a gibberellin (GA) biosynthetic inhibitor, abolished the FR promoted epicotyl elongation, indicating a role for GAs in this process. There was no significant difference in epicotyl elongation rates of R plus FR irradiated explants treated with GA(1) or GA(20) and R irradiated explants treated with GA(1). However, R irradiation inhibited subsequent epicotyl elongation of GA(20) treated explants. Moreover, the observation, using GC-MS, that GA(1) and GA(20) are native GAs in cowpea lends support to the concept that phytochrome may control the conversion of endogenous GA(20) to GA(1) in cowpea.


Acetohydroxyacid synthase isozyme I from Escherichia coli has unique catalytic and regulatory properties.
AHAS I is an isozyme of acetohydroxyacid synthase which is apparently unique to enterobacteria. It has been known for over 20 years that it has many properties which are quite different from those of the other two enterobacterial AHASs isozymes, as well as from those of "typical" AHASs which are single enzymes in a given organism. These include a unique mechanism for regulation of expression and the absence of a preference for forming acetohydroxybutyrate. We have cloned the two subunits, ilvB and ilvN, of this Escherichia coli isoenzyme and examined the enzymatic properties of the purified holoenzyme and the enzyme reconstituted from purified subunits. Unlike other AHASs, AHAS I demonstrates cooperative feedback inhibition by valine, and the kinetics fit closely to an exclusive binding model. The formation of acetolactate by AHAS I is readily reversible and acetolactate can act as substrate for alternative AHAS I-catalyzed reactions.


[A case of intrasellar meningioma with panhypopituitarism and hyperprolactinemia].
A case of intrasellar meningioma is reported. A 49-year-old woman was admitted to our hospital on July 22, 1985, complaining of reduced visual acuity and visual field defect. Visual acuity was 0.6 in the right eye and 0.1 in the left eye. Visual field examination revealed upper temporal quadrantanopsia on the right side and incomplete temporal hemianopsia on the left side. Ocular fundi were normal. X-ray films of the skull showed a balloon-shaped sella turcica with "double floor". CT scan showed a isodense mass with central low density occupying the intrasellar and suprasellar region. After administration of contrast medium, almost homogenous enhancement was noted. Bilateral carotid angiographies demonstrated that horizontal portion of the right anterior cerebral artery was raised. No tumor blush was evident. Endocrinologic function tests confirmed a complete deficit of the anterior lobe hormones except for elevated serum prolactin level of 110 ng/ml. She showed no galactorrhea. On August 6, 1985, the sella turcica was reached via the transsphenoidal rhinoseptal approach. The sella floor and dura matter were intact. The grey, soft and necrotic tumor tissue was encountered and bleeding was controllable. The tumor extending to suprasellar region was firm in consistency. Pathologically, the tumor was a typical meningothelial meningioma. Postoperatively, visual field defect improved and visual acuity was recovered on the right side immediately. Postoperative CT scan showed a thin residual enhanced lesion, which was the attachment of the tumor. It seemed to be the elevated diaphragma sellae. Clinical observation, radiological and endocrinological findings of intrasellar meningioma are similar to that of non-functioning pituitary adenoma.(ABSTRACT TRUNCATED AT 250 WORDS)


Electrotransfer of fixed phosphoproteins from pieces of dried polyacrylamide gel to small disks of nitrocellulose, nylon or polyvinylidene difluoride.
A simple method for the transfer of 32P-labeled proteins from dried polyacrylamide gels to small disks of nitrocellulose, nylon or polyvinylidene difluoride (PVDF) is described. Gel pieces containing the desired phosphoprotein are rehydrated in buffer containing sodium dodecyl sulfate (SDS) and sealed in agarose in a glass tube over a supporting gel of polyacrylamide. Protein is transferred upwards through a discontinuous density gradient of SDS-buffer and methanol to a disk of membrane sealed to the mouth of the tube with dialysis membrane. The method allows the concentration of a phosphoprotein present in several gel pieces to a single disk of immobilized membrane. Recovery of phosphoprotein was at least as good as obtained with conventional electroblotting. Application of the method to the analysis of the phosphoamino acid content of the astrocyte marker, glial fibrillary acidic protein, is described.


Detection of Mycobacterium avium subsp. paratuberculosis in drinking water and biofilms by quantitative PCR.
It has been suggested that Mycobacterium avium subspecies paratuberculosis has a role in Crohn's disease. The organism may be acquired but is difficult to culture from the environment. We describe a quantitative PCR (qPCR) method to detect M. avium subsp. paratuberculosis in drinking water and the results of its application to drinking water and faucet biofilm samples collected in the United States.


State policies and the financing of acquired immunodeficiency syndrome care.
State policies, with respect to the operation of Medicaid programs and the regulation of private health insurance, affect who gets what care, how much is spent, and who ultimately pays. A RAND Corporation study was used to assess States and the District of Columbia in terms of the effects of their Medicaid and health insurance regulations on people with acquired immunodeficiency syndrome and other human immunodeficiency virus-related illnesses. State characteristics are used to explain the individual State policy rankings.


Design of an ultra-thin near-eye display with geometrical waveguide and freeform optics.
Small thickness and light weight are two important requirements for a see-through near-eye display which are achieved in this paper by using two advanced technologies: geometrical waveguide and freeform optics. A major problem associated with the geometrical waveguide is the stray light which can severely degrade the display quality. The causes and solutions to this problem are thoroughly studied. A mathematical model of the waveguide is established and a non-sequential ray tracing algorithm is developed, which enable us to carefully examine the stray light of the planar waveguide and explore a global searching method to find an optimum design with the least amount of stray light. A projection optics using freeform surfaces on a wedge shaped prism is also designed. The near-eye display integrating the projection optics and the waveguide has a field of view of 28°, an exit pupil diameter of 9.6mm and an exit pupil distance of 20mm. In our final design, the proportion of the stray light energy over the image output energy of the waveguide is reduced to 2%, the modulation transfer function values across the entire field of the eyepiece are above 0.5 at 30 line pairs/mm (lps/mm). A proof-of-concept prototype of the proposed geometrical waveguide near-eye display is developed and demonstrated.


[Rare blood group B (A) detection and safe transfusion].
This study was aimed to investigate the genetic characteristics, identification method and transfusion strategy of rare blood type B(A). The rare blood group B(A) was typed by serological technique, PCR-SSP genotyping and sequencing of exon 6, 7 of ABO blood group. The genetic characteristics and molecular mechanism of B(A) blood group were also analyzed. Blood group compatibility test was conducted between blood donors of B(A) and recipients by clinical transfusion. The results showed that both forward and reverse grouping did not match the 3 cases of serological result in their family survey, while all of the 3 cases were grouped as AB blood group by forward grouping, B blood group by reverse grouping with serological result and B(A)04/001 group were genotyped by ABO genotyping. The patient of B blood group was transfused by 1 bag of washed red blood cells of donor of B(A) under closely monitoring, the patient's condition changed, and a mild adverse transfusion reaction was appeared. Washed red blood cell of O blood group was transfused into B(A) patient without blood transfusion reaction. It is concluded that the forward ABO serological grouping and reverse ABO serological grouping are not compatible, that may be verified by family survey and molecular biological methods. If in some cases transfusion therapy was applied, and group B(A) can not be transfused to the patient with group B or AB. Thus, transfusion compatibility or autologous transfusion can be adopted to transfuse to the patient from group B(A).


[miR-451 inhibits malignant progression of multiple myeloma RPMI-8226 cells by targeting c-Myc].
Objective: To investigate the effects of microRNA-451 on proliferation, invasion and migration of multiple myeloma RPMI-8226 cells and its mechanism. Methods: RPMI-8226 cells cultured in vitro were divided into blank control group (untransfected), negative control (NC) group and miR-451 mimic transfected (miR-451) group. The expression of miR-451 was detected by real-time quantitative PCR (qRT-PCR), cell proliferation was detected by 3-(4, 5-dimethyl-2-thiazolyl)-2, 5-diphenyl-2-H-tetrazolium bromide (MTT) array and clone formation experiment, cell invasion and migration were detected by Transwell, and the expressions of c-Myc, MMP-2 and MMP-9 proteins were detected by western blot. The targeting relationship between miR-451 and c-Myc was detected by double luciferase reporter gene assay. Results: Compared to the blank control group, the expression level of miR-451 was increased (2.85±0.27 vs 1.02±0.06), while the cell survival rate [(47.28±3.15)% vs (93.65±6.52)%], cloning formation rate [(15.03±1.34)% vs (28.48±2.12)%], invasive cell number (86.65±5.58 vs 135.47±9.85), migrating cell number (106.36±6.48 vs 165.28±11.05) and the expression level of c-Myc(0.35±0.03 vs 0.66±0.05), MMP-2 (0.20±0.02 vs 0.48±0.03) and MMP-9 (0.28±0.03 vs 0.59±0.06) protein were significantly decreased in the miR-451 group (P<0.05). In the negative control group, the expression level of miR-451, cell viability, clone formation rate, invasive cell number, migrating cell number, c-Myc protein, MMP-2 protein and MMP-9 protein were 0.94±0.05, (95.16±5.04)%, (27.55±2.26)%, (128.96±8.32) and (158.65±8.76), 0.68±0.06, 0.51±0.03, 0.54±0.03, respectively. There were no significant differences between the blank control group and the NC group (P>0.05). Double luciferase reporter gene experiment confirmed that c-Myc was a potential target gene of miR-451. Conclusion: miR-451 can inhibit the proliferation, invasion and migration of RPMI-8226 cells by targeting c-Myc.


Comparative study of efficiency of dental methods for identification of burn victims in two bus accidents in Spain.
This article deals with the usefulness and limitations of dental identification methods of human burn victims of two buses accidents in Spain. The first accident happened in Bailen in 1996 and involved 28 badly burned Spanish victims. In the second, in Illescas in 1997, 8 Japanese and 2 Spanish victims lost their lives. In both accidents, postmortem forensic procedures for identification were used, including external and internal examination, routine photographs, and dental examination. Dental identification was established in 57% of the cases in the Bailen accident and 80% in the Illescas accident. The success rate of dental identification varies considerably depending on the nature of the accident, the nationality and country of residence of the victims, the incidence of dental treatment, the availability of adequate dental records, and the degree of dental injuries. A discussion of procedures and methods for identification recommended when dealing with burn victims is included.


Carboranyl oligonucleotides: 4. synthesis and physicochemical studies of oligonucleotides containing 2'-O-(o-carboran-1-yl)methyl group.
Boronated oligonucleotides are potential candidates for antisense oligonucleotide technology (AOT), boron neutron capture therapy (BNCT), and as tools in molecular biology. A method was developed for the solid phase synthesis of oligonucleotides containing 2'-O-(o-carboran-1-yl-methyl) (2'-CBM) group. Synthesis was performed using a standard beta-cyanoethyl cycle and automated DNA synthesizer. Manual steps were performed for the insertion of a modified monomer bearing the 2'-CBM group. Several tetradecanucleotides complementary to DNA-HCMV, and bearing 2'-CBM modification near the 3'-end or 5'-end or in the middle of the oligonucleotide chain were synthesized. The resulting oligomers were characterized by polyacrylamide gel electrophoresis (PAGE), reverse phase high-performance liquid chromatography (RP-HPLC), matrix-assisted laser desorption ionization mass spectrometry (MALDI-MS) and ultraviolet spectroscopy (UV), circular dichroism (CD), and melting temperature (Tm) measurements. Tm of duplexes formed between 2'-CBM-modified tetradecanucleotides and complementary DNA and RNA template were compared with those formed by the unmodified oligonucleotide and complementary sequence. The stability of 2'-CBM oligonucleotides in the presence of phosphodiesterase I from Crotalus atrox venom and in human serum was studied. Oligonucleotides bearing the 2'-CBM group are characterized by increased resistance to enzymatic digestion, increased lipophilicity, and the ability to form stable duplexes with complementary templates.


Plasma kallikrein-mediated activation of the renin-angiotensin system does not require prior acidification of prorenin.
Activation of prorenin in the neutral phase after pH 3.3 dialysis of human plasma depends on clotting factor XII-initiated prekallikrein to kallikrein conversion. Acid dialysis may be necessary for destroying kallikrein inhibitors or rendering prorenin susceptible to attack by kallikrein. If the latter possibility proves true, it is difficult to see how the factor XII-kallikrein pathway could activate prorenin in vivo. Plasma prorenin was therefore separated from active renin and from the protease inhibitors alpha 2-macroglobulin, C1-inactivator, alpha 1-antitrypsin, inter-alpha-trypsin inhibitor, and antithrombin III by gel filtration on Sephadex G-100 and affinity chromatography on Blue Sepharose CL-6B at neutral pH. The resulting prorenin preparation could be activated at pH 7.5 by highly purified human plasma kallikrein, which was prepared from prekallikrein by activation with active factor XII fragment beta-factor XII a. Activation proceeded at 4 and 37 C at a kallikrein concentration of 2 micrograms/ml, which is approximately 5% of the prekallikrein concentration in normal plasma. It appears that an acid-induced conformational change of the prorenin molecule is not required for its activation by plasma kallikrein.


A β-N-acetylhexosaminidase Amuc_2109 from Akkermansia muciniphila protects against dextran sulfate sodium-induced colitis in mice by enhancing intestinal barrier and modulating gut microbiota.
Inflammatory bowel disease (IBD) is associated with the microbial composition of the gut and its metabolites. Akkermansia muciniphila is a probiotic that exerts a significant alleviative or therapeutic effect on host enteritis. This study was designed to determine the protective effect and potential mechanism underlying the secretion of β-acetylaminohexosidase (Amuc_2109) by A. muciniphila against dextran sulfate sodium (DSS)-induced colitis in mice. C57BL/6 mice were gavaged with Amuc_2109 for 21 days, and during the last seven days of treatment, they drank DSS dissolved in their drinking water to induce colitis. Our results showed that supplementation with Amuc_2109 improved DSS-induced colitis as evidenced by lowered disease activity index (DAI) scores, reduced weight loss, increased colon length, and inhibited oxidative stress. In addition, Amuc_2109 inhibited the overexpression of inflammatory cytokines (TNF-α, IL-1β, IL-6) and the NLR family pyrin domain containing 3 (NLRP3) inflammasome in DSS-induced colitis. Furthermore, supplementation with Amuc_2109 also restored the mRNA expression of tight junction proteins (ZO-1, occludin, claudin-1). Further analysis of fecal microbial 16S rRNA sequences showed that Amuc_2109 reshaped the intestinal microbiota. While the anti-inflammatory effects of Amuc_2109 were only manifested with the wild-type protein, the anti-inflammatory effects were completely lost after the mutation of its key catalytic amino acids rendered Amuc_2109 inactive. In summary, these findings demonstrate the potential of Amuc_2109, as a therapeutic agent for ulcerative colitis. We posit that it will provide additional assistance in the prevention and treatment of mucus layer-related diseases such as ulcerative colitis.


Education for professional nursing practice: looking backward into the future.
The publication of the 1965 ANA position paper calling for the baccalaureate degree as the minimum requirement for entry into professional nursing practice initiated an impassioned debate which continues to frustrate and divide nursing. An examination of the entry into practice debate from an educational perspective reveals some interesting similarities in the social and political pressures felt by nursing in 1965 and the challenges that exist today. In looking back at the course of nursing education during the past thirty-seven years, some useful insights emerge which can help to set direction for the future. The author argues that it is time to leave the old debate behind and agree on the importance of developing a better educated profession.


"Oxygen supply" as modulator of aging processes: hypoxia and hyperoxia models for aging studies.
Cell growth is regulated by several factors, including oxygen supply, which influence cell metabolism. Aging is characterized by decreased oxygen supply to tissue, a reduction of tissue PO(2) and of the activity of several enzymes and metabolic factors. The oxygen-gradient diffusion at capillary tissue level is essential for the cellular survival, while the homeostasis of the oxygen in the arterial blood is mediated by reflexes sensitive to oxygen decrease and by release of several factors. Aging is correlated with a reduction of cells' oxygen supply concomitant to a parallel decrease in oxygen demand by tissues. Both chronic hypoxia or hyperoxia are considered as stresses. Indeed, in both conditions, free radical species, which damage structural and functional components of the membrane, are generated. ROS (reactive oxygen species) are physiological products of aerobic life and their accumulation affects aging. Because hypoxia per se modulates mitochondria activity, influencing oxygen consumption, hypoxia and aging could share some link. Moreover, the observation that in hypoxia or hyperoxia there is an accumulation of lipofucsine as a general reaction to stress is consistent with the accumulation of such components during aging. Correlation between hypoxia-hyperoxia and life-span remains open until we solve the question of how and why do cells sense oxygen. In other words, to better understand aging we need to know what O(2) species are being sensed by cells. In conclusion, hypoxia and hyperoxia represent an experimental model adequate for studying aging processes.


Survival of cardiac allograft in highly sensitized and nonsensitized rats treated with FK506.
The effect of FK506 was studied in sensitized and nonsensitized recipients using a rat cardiac allograft model. Heart grafts from Male ACI rats were heterotopically transplanted to Male LEW rats. The sensitized rat models were established by using donor type blood, admixed with immunoadjuvant (Adjuvant Complete Freund), seven days prior to transplantation. FK506 was administrated from day 0 to day 14 posttransplantation. The results showed that FK506 could prolong allograft survival in both sensitized and nonsensitized rat recipients in a dose-dependent manner, although, the dose required in sensitized recipients was higher than in nonsensitized recipients. The minimal effective dose of FK506 was 0.32 mg/kg/day in sensitized recipients and 0.1 mg/kg/day in nonsensitized recipients.


Surveillance Colonoscopy after Appendicectomy in Patients over the Age of 40: Targeted Audit of Outcomes and Variability in Practice.
Introduction: Recent evidence suggests the need to proceed with a surveillance colonoscopy in patients above the age of 40 years who undergo appendicectomy for acute appendicitis, given the higher risk of an underlying colonic tumor. After anecdotally observing a substantial variability in terms of adaptation of these recommendations by the on-call surgical teams, we performed a clinical audit regarding our relevant endoscopic follow-up compliance rates to identify areas for improvement of our practise. Materials Methods: We performed a retrospective review of the electronic records of all patients above 40 years who had appendicectomy for acute appendicitis within a 3-year period in our institution, assessing as primary outcome the actual performance of a follow-up colonoscopy and the detected endoscopic findings. Results: Our results demonstrated that more than 80% of our patients did not have an endoscopic follow-up, as suggested by the current evidence. In addition, with respect to the subspecialisation of the parent surgical team, it seems that non-colorectal teams had lower compliance regarding the arrangement of endoscopic surveillance, when compared to specialist colorectal team. Conclusions: Emergency surgical teams need to be further educated with respect to the current practise recommendations concerning the appropriate endoscopic follow-up after the performance of appendicectomy for acute appendicitis. Establishment of dedicated bundles of postoperative care, as well as clear relevant guidance from the gastrointestinal/emergency surgery societies would be of great value in this direction.


Effect of Ferredoxin on the Diaphorase Activity of Cyanobacterial Ferredoxin-NADP Reductase.
The interaction of ferredoxin-NADP reductase (FNR) and ferredoxin (Fd) results in an enhanced rate of reaction and a shift of the pH optimum for the FNR-mediated diaphorase reaction. Low concentrations of NaCl (<100 millimolar), favorable for formation of the FNR:Fd complex, further magnify the alteration of the diaphorase reaction; the activity is enhanced 3-fold and pH optimum is shifted from 9.5 to 7.8. The Fd-stimulated diaphorase activity of FNR may result either from a conformational change of the enzyme and/or from a transition from a two electron to a one electron reaction.


Genomic structure and promoter analysis of the human obese gene.
The human gene encoding the homolog of the mouse obese (ob) gene was isolated and partially characterized. The human ob gene consists of three exons and two introns and spans about 18 kilobase pairs (kb), encoding a 3.5-kb cDNA. A 3-kb 5'-flanking region of the gene was cloned and transient transfection assay with luciferase reporter confirmed the promoter activity in differentiated F442-A adipocytes. Potential regulatory elements are discussed in this report.


New mutant rabbit strain with hypercholesterolemia and atherosclerotic lesions produced by serial inbreeding.
We have investigated the characteristics of the Kurosawa and Kusanagi hypercholesterolemic (KHC) rabbit, a strain manifesting inherited and persistent hypercholesterolemia and produced by inbreeding from mutants discovered in 1985. These KHC rabbits had abnormally high serum cholesterol, triglyceride, and phospholipid concentrations 8 to 10 times greater than the concentrations in clinically normal Japanese white rabbits (Oryctolagus cuniculus). The KHC rabbits also had decreased serum high-density lipoprotein cholesterol concentration, about one-third the value in clinically normal rabbits. In addition, the serum lipoprotein electrophoretic patterns were characterized by a strong, broad beta-lipoprotein band and a diminished alpha-lipoprotein band. Fractionation of lipoprotein lipids revealed increased cholesterol, phospholipid, and triglyceride in the low-density lipoprotein fraction; increased cholesterol and phospholipid in the very low-density lipoprotein fraction; and decreased cholesterol and triglyceride in the high-density lipoprotein fraction. On the basis of incidence of hypercholesterolemia in mating experiments, this abnormality is thought to have been inherited as a single autosomal recessive gene mutation, and analysis of the low-density lipoprotein receptor indicated that the KHC rabbit has a 12-base pair deletion in the low-density lipoprotein receptor mRNA. Macroscopic analysis of the aorta revealed the atheromatous lesions at 2 months of age, drastically increased lesional areas in the total aortic surface at 8 months of age, and a high incidence of coronary atheromas and xanthomas. Light microscopy revealed two typical lesions in the aorta fatty plaque and medical calcification.(ABSTRACT TRUNCATED AT 250 WORDS)


A Systematic Review of Head and Neck Cancer Health Disparities: A Call for Innovative Research.
(1) Describe the existing head and neck cancer health disparities literature. (2) Contextualize these studies by using the NIMHD research framework (National Institute on Minority Health and Health Disparities). (3) Explore innovative ideas for further study and intervention. Ovid MEDLINE, Embase, Web of Science, and Google Scholar. Databases were systematically searched from inception to April 20, 2020. The PRISMA checklist was followed (Preferred Reporting Items for Systematic Reviews and Meta-analyses). Two authors reviewed all articles for inclusion. Extracted data included health disparity population and outcomes, study details, and main findings and recommendations. Articles were also classified per the NIMHD research framework. There were 148 articles included for final review. The majority (n = 104) focused on health disparities related to at least race/ethnicity. Greater than two-thirds of studies (n = 105) identified health disparities specific to health behaviors or clinical outcomes. Interaction between the individual domain of influence and the health system level of influence was most discussed (n = 99, 66.9%). Less than half of studies (n = 61) offered specific recommendations or interventions. There has been extensive study of health disparities for head and neck cancer, largely focusing on individual patient factors or health care access and quality. This review identifies gaps in this research, with large numbers of retrospective database studies and little discussion of potential contributors and explanations for these disparities. We recommend shifting research on disparities upstream toward a focus on community and societal factors, rather than individual, and an evaluation of interventions to promote health equity.


The Reliability and Validity of Kiddie-Schedule for Affective Disorders and Schizophrenia - Present and Life-time Version - Persian Version.
The validity and reliability of a Persian version of the Kiddie Schedule for Affective Disorders and Schizophrenia-Present and Lifetime Version (K-SADS-PL-P) was evaluated. The K-SADS-PL-P was administered to 102 inpatients (mean age = 15.3 yrs, SD = 1.81) in a child and adolescent psychiatric ward. The psychometric properties were evaluated in comparison to the results of clinical diagnosis. The K-SADS-PL-P showed good-to-excellent concurrent validity in diagnosing current major disorders. Test-retest reliabilities of most of the current diagnoses were also good to excellent. The Persian version of the K-SADS-PL provides reliable and valid youth psychiatric diagnoses.


Mechanical and Electrochemical Performance of Carbon Fiber Reinforced Polymer in Oxygen Evolution Environment.
Carbon fiber-reinforced polymer (CFRP) is recognized as a promising anode material to prevent steel corrosion in reinforced concrete. However, the electrochemical performance of CFRP itself is unclear. This paper focuses on the understanding of electrochemical and mechanical properties of CFRP in an oxygen evolution environment by conducting accelerated polarization tests. Different amounts of current density were applied in polarization tests with various test durations, and feeding voltage and potential were measured. Afterwards, tensile tests were carried out to investigate the failure modes for the post-polarization CFRP specimens. Results show that CFRP specimens had two typical tensile-failure modes and had a stable anodic performance in an oxygen evolution environment. As such, CFRP can be potentially used as an anode material for impressed current cathodic protection (ICCP) of reinforced concrete structures, besides the fact that CFRP can strengthen the structural properties of reinforced concrete.


Langmuir films from antibodies based on amphiphilic polyelectrolytes.
A technique for forming Langmuir films from antibodies based on an amphiphilic polyelectrolyte was developed. The physicochemical and immunochemical properties of the Langmuir films obtained were studied. The interaction of HBsAg with the films was found to be described by a model with one binding site, whereas that of HBsAg with antibodies adsorbed on a polystyrene plate, by a model with a positive cooperativity. The use of the novel Langmuir films from antibodies increases the sensitivity of the immunoenzyme assay.


Interactions between inositol trisphosphate receptors and fluorescent Ca2+ indicators.
Fura-2 and BAPTA were previously shown to be competitive antagonists of inositol trisphosphate (InsP3) receptors, but for practical reasons the analyses were performed at pH 8.3. We recently developed a scintillation proximity assay (SPA) for pure cerebellar InsP3 receptors which allows low affinity interactions to be characterized and is readily applicable to scarce or expensive ligands. In the present study, we use SPA to demonstrate that at pH 7.2, many of the commonly used fluorescent Ca2+ indicators reversibly displace 3H-InsP3 from its receptor and that they differ substantially in their affinities for the InsP3 receptor (IC50 = 6.5-137 microM). Recombinant type 1 InsP3 receptors expressed in Sf9 cells were used to examine 3H-InsP3 binding in cytosol-like medium: both fura-2 (IC50 = 796 +/- 86 microM) and Ca Green-5N (IC50 = 62 +/- 7 microM) completely inhibited the binding, but only in their Ca(2+)-free forms. Similar results were obtained with type 3 InsP3 receptors. We conclude that many Ca2+ indicators in their Ca(2+)-free forms compete with InsP3 for binding to its receptor, and that for Ca Green-5N the interaction occurs with sufficient affinity to significantly perturb physiological responses.


Chemical diversity in Himalayan Elsholtzia species.
The terpenoid composition of essential oils from the leaves of five Elsholtzia species, viz., E. eriostachya Benth., E. cristata Willd., E. polystachya Benth., E. flava Benth., and E. pilosa Benth., collected from the Himalayan region (India), was examined by GC, GC/MS, and NMR analyses. Comparison of the results with previous reports revealed new chemotypes. Cluster analysis was carried out in order to discern the similarities and differences within the essential-oil compositions at their subspecies/chemotype level. Based on the major constituents of the essential oils, six chemical groups were obtained.


[Modulation of anthracycline resistance by reserpine in P388 leukemia cells].
The activity of reserpine and a possible mechanism by which it reverses the resistance to both doxorubicin and pirarubicin in doxorubicin-resistant P388 leukemia (P388/DOX) cells were examined in vitro. During 48 hr drug-exposure, the sensitivity of doxorubicin and pirarubicin were potentiated markedly when reserpine was present at the concentration of 1 microgram/ml, which is not toxic to P388 leukemia (P388/S) cells. However, reserpine had little effect on the cytotoxicity of doxorubicin and pirarubicin in the sensitive parent cell. Reserpine at 0.5-20 micrograms/ml increased intracellular accumulation of doxorubicin and pirarubicin in the drug-resistant cells. The potentiating action of reserpine was stronger when the cells were preincubated with reserpine within 30 min. Efflux of doxorubicin and pirarubicin was greater in drug-resistant cells compared to sensitive cells. This enhanced efflux of drug resulted in a decrease in the intracellular accumulation of doxorubicin in the drug-resistant cells. When the resistant cells were exposed to 2 micrograms/ml of reserpine, this enhanced efflux was blocked. A similar effect of reserpine on doxorubicin was seen with the efflux pattern of pirarubicin. From the measurements of drug uptake and efflux, it seems that like other multiple drug resistance modifiers, reserpine modulates anthracycline resistance by increasing intracellular accumulation of drug.


11β-hydroxysteroid dehydrogenases: intracellular gate-keepers of tissue glucocorticoid action.
Glucocorticoid action on target tissues is determined by the density of "nuclear" receptors and intracellular metabolism by the two isozymes of 11β-hydroxysteroid dehydrogenase (11β-HSD) which catalyze interconversion of active cortisol and corticosterone with inert cortisone and 11-dehydrocorticosterone. 11β-HSD type 1, a predominant reductase in most intact cells, catalyzes the regeneration of active glucocorticoids, thus amplifying cellular action. 11β-HSD1 is widely expressed in liver, adipose tissue, muscle, pancreatic islets, adult brain, inflammatory cells, and gonads. 11β-HSD1 is selectively elevated in adipose tissue in obesity where it contributes to metabolic complications. Similarly, 11β-HSD1 is elevated in the ageing brain where it exacerbates glucocorticoid-associated cognitive decline. Deficiency or selective inhibition of 11β-HSD1 improves multiple metabolic syndrome parameters in rodent models and human clinical trials and similarly improves cognitive function with ageing. The efficacy of inhibitors in human therapy remains unclear. 11β-HSD2 is a high-affinity dehydrogenase that inactivates glucocorticoids. In the distal nephron, 11β-HSD2 ensures that only aldosterone is an agonist at mineralocorticoid receptors (MR). 11β-HSD2 inhibition or genetic deficiency causes apparent mineralocorticoid excess and hypertension due to inappropriate glucocorticoid activation of renal MR. The placenta and fetus also highly express 11β-HSD2 which, by inactivating glucocorticoids, prevents premature maturation of fetal tissues and consequent developmental "programming." The role of 11β-HSD2 as a marker of programming is being explored. The 11β-HSDs thus illuminate the emerging biology of intracrine control, afford important insights into human pathogenesis, and offer new tissue-restricted therapeutic avenues.


Developmental exposure to the SSRI citalopram causes long-lasting behavioural effects in the three-spined stickleback (Gasterosteus aculeatus).
Selective Serotonin re-uptake inhibitors (SSRIs) are a class of psychotropic drugs used to treat depression in both adolescents and pregnant or breast-feeding mothers as well as in the general population. Recent research on rodents points to long-lasting behavioural effects of pre- and perinatal exposure to SSRIs which last into adulthood. In fish however, studies on effects of developmental exposure to SSRIs appears to be non-existent. In order to study effects of developmental SSRI exposure in fish, three-spine sticklebacks were exposed to 1.5 µg/l of the SSRI citalopram in the ambient water for 30 days, starting two days post-fertilisation. After approximately 100 days of remediation in clean water the fish were put through an extensive battery of behavioural tests. Feeding behaviour was tested as the number of bites against a piece of food and found to be increased in the exposed fish. Aggression levels were measured as the number of bites against a mirror image during 10 min and was also found to be significantly increased in the exposed fish. Novel tank behaviour and locomotor activity was tested in an aquarium that had a horizontal line drawn half-way between the bottom and the surface. Neither the latency to the first transition to the upper half, nor the number of transitions or the total time spent in the upper half was affected by treatment. Locomotor activity was significantly reduced in the exposed fish. The light/dark preference was tested in an aquarium where the bottom and walls were black on one side and white on the other. The number of transitions to the white side was significantly reduced in the exposed fish but there was no effect on the latency to the first transition or the total time spent in the white half. The results in the current study indicate that developmental SSRI exposure causes long-lasting behavioural effects in fish and contribute to the existing knowledge about SSRIs as environmental pollutants.


First Report of Impatiens Necrotic Spot Virus Infecting Lettuce in Arizona and Southern Desert Regions of California.
Impatiens necrotic spot virus (INSV; family Tospoviridae, genus Orthotospovirus) is a thrips-borne pathogen that infects a wide range of ornamental and vegetable crops. INSV was first reported in lettuce (Lactuca sativa) in the Salinas Valley of CA (Monterey County) in 2006 (Koike et al. 2008). Since then, the pathogen has continued to impact lettuce production in the region, causing severe economic losses with increasing incidence and severity in recent years. Tomato spotted wilt virus (TSWV), another tospovirus, also infects lettuce, but its occurrence is much less frequent than INSV (Kuo et al. 2014). While INSV has not been reported in the desert areas of CA and AZ, there are concerns that the virus could become established in this region. In early March 2021, symptoms resembling those caused by orthotospovirus infection were observed in several romaine and iceberg lettuce fields in the Yuma and Tacna regions of Yuma County, AZ. Symptoms included leaves that exhibited tan to dark brown necrotic spots, distorted leaf shapes, and stunted plant growth. Similar symptoms were also reported in romaine fields and one green leaf and iceberg lettuce field in the neighboring Imperial and Riverside Counties of CA. A total of 14 samples (5 from Tacna, 4 from Yuma, 4 from Imperial, 1 from Riverside) were tested using ImmunoStrips (Agdia, Elkhart, IN) for INSV and TSWV. Results confirmed the presence of INSV in 13 out of 14 samples, and the absence of INSV in one sample originating from Yuma. All 14 samples tested negative for TSWV. The 13 INSV positive samples were processed for RT-PCR validation. Total RNA was extracted from each sample using the RNeasy Plant Mini Kit (Qiagen, Valencia, CA). RT-PCR was performed with OneStep Ahead RT-PCR Kit (Qiagen) with primers to the N gene of INSV S RNA (Accession KF745140.1; INSV F = CCAAATACTACTTTAACCGCAAGT; INSV R = ACACCCAAGACACAGGATTT). All reactions generated a single amplicon at the correct size of 524 bp. One sample each from Yuma, Tacna, and Brawley (Imperial County), as well as a romaine lettuce sample collected from the Salinas Valley in March 2021, were sent for Sanger bi-directional sequencing (Eton Biosciences, San Diego, CA). Sequence analysis revealed that all three desert samples (Yuma, Tacna, and Brawley with Accessions OK340696, OK340697, OK340698, respectively) shared 100% sequence identity and 99.43% identity to the Salinas Valley 2021 sample (SV-L2, Accession OK340699). Additionally, all desert samples shared 99.24% sequence identity to the Salinas Valley lettuce isolate previously described in 2014 (SV-L1, Accession KF745140.1; Kuo et al. 2014), while the SV-L2 and SV-L1 sequences shared 99.43% identity. By the end of the season (April 2021) a total of 43 lettuce fields in Yuma County, AZ, and 9 fields in Imperial and Riverside Counties, CA were confirmed to have INSV infection using ImmunoStrips. Impacted fields included romaine, green leaf, red leaf, and head lettuce varieties, and both direct-seeded and transplanted lettuce, under conventional and organic management regimes. In AZ, INSV incidence in fields ranged between 0.2% and 33%, while in Imperial and Riverside Counties, CA, field incidence remained low at less than 0.1%. It is possible that INSV was introduced from the Salinas Valley of CA through the movement of infected lettuce transplants and/or thrips vectors. To our knowledge, this is the first report of INSV infecting lettuce in Arizona and the southern desert region of California.


Rituximab in pemphigus.
Pemphigus is a severe autoimmune blistering disease mediated by pathogenic anti-desmoglein antibodies leading to an inter keratinocyte disjunction. Rituximab is a monoclonal antibody that binds to the CD-20 antigen of B lymphocytes, which causes B-cell depletion and a subsequent reduction in pathogenic autoantibodies. Its therapeutic role in pemphigus has been progressively growing with increasing evidence of successful outcomes. Rituximab was initially off-labeled used as an alternative in patients with recalcitrant or relapsing pemphigus and in patients with contraindications to systemic corticosteroids. Recently, a large randomized clinical trial has shown that first-line use of rituximab combined with short-term prednisone regimen was both more effective and potentially safer than a standard regimen of high doses of corticosteroids in patients with moderate to severe pemphigus.


Contribution of HLA-DRB1 * 09: 01 allele to development of minocycline induced antineutrophil cytoplasmic antibody (ANCA)-associated cutaneous vasculitis: report of two cases.
We report two cases of antineutrophil cytoplasmic antibody (ANCA)-associated vasculitis (AAV) that developed after long-term oral administration of minocycline and consider the contribution of human leukocyte antigen (HLA)-DRB 1 * 09: 01 allele to its development. Case 1; A 47-year-old man receiving minocycline for palmoplantar pustulosis for 24 months developed fever, arthralgia, and irregular livedo on the bilateral lower legs. Skin biopsy demonstrated vasculitis, while a blood test showed positivity of myeloperoxidase (MPO)-ANCA. Discontinuation of minocycline and oral administration of prednisolone relieved the symptoms promptly. Case 2; A 53-year-old woman developed reddish-brown livedo reticularis with tenderness on the bilateral lower legs after administration of minocycline to treat palmoplantar pustulosis for 24 months. Although skin biopsy did not demonstrate vasculitis, a blood test showed MPO-ANCA positivity. Cessation of minocycline resulted in rapid improvement of the cutaneous lesions and constitutional symptoms. We diagnosed both cases as having Drug-associated ANCA-associated Vasculitis (DAV) caused by minocycline according to the diagnostic criteria proposed by Cluver et al. Further examination revealed the presence of HLA-DRB1 * 09:01 allele in both cases. This allele has been implicated in the genetic background of idiopathic microscopic polyangiitis (MPA) in the Japanese population. Our finding suggests a relationship between the development of MPO-ANCA or DAV caused by minocycline and HLA-DRB1 * 09:01 allele, but will have to confirmed by further studies with larger numbers of patients.


Enzymatic Precipitation Enhanced Surface Plasmon Resonance Immunosensor for the Detection of Salmonella in Powdered Milk.
Contamination of food by pathogenic bacteria has always been a serious threat for human health. The amount of food exports and imports has been increasing in recent years which requires precise food quality control with short analysis time and simplified sample treatment. Surface plasmon resonance (SPR) immunosensor enhanced by biocatalyzed precipitation was developed for the analysis of Salmonella in dairy products. The specific capture antibody was immobilized on the SPR chip which allowed a direct label-free detection of Salmonella Typhimurium with the limit of detection (LOD) of 104 CFU·mL-1 and the analysis time of 10 min. Alternatively, the secondary detection antibody was conjugated with horseradish peroxidase to provide a signal enhancement by the biocatalyzed conversion of 4-chloro-1-naphthol to insoluble benzo-4-chlorocyclohexadienone. The formation of precipitate was studied in detail by atomic force microscopy (AFM). The sensitivity was increased 40 times in case of the precipitation-enhanced detection compared to the label-free approach. The optimized method provided LOD of 100 CFU·mL-1 with linear range up to 106 CFU·mL-1. The total time of analysis including bacteria binding and enhancement step was below 60 min. The capability to analyze real samples with complex matrices was demonstrated on the detection of Salmonella in powdered milk. The developed sensor represents simple and robust approach for routine monitoring of food contamination.