Culture medium from TNF-α-stimulated mesenchymal stem cells attenuates allergic conjunctivitis through multiple antiallergic mechanisms.
The immunomodulatory and anti-inflammatory functions of mesenchymal stem cells (MSCs) have been demonstrated in several autoimmune/inflammatory diseases, but their contribution to allergic conjunctivitis and underlying antiallergic mechanisms remain elusive. We sought to explore the clinical application of MSCs to experimental allergic conjunctivitis (EAC) and its underlying antiallergic mechanisms. Culture medium from TNF-α-stimulated, bone marrow-derived MSCs (MSC-CMT) was administered topically to mice with EAC, and the related allergic symptoms and biological changes were evaluated. Murine spleen-derived B cells, bone marrow-derived mast cells (MCs), and lung vascular endothelial cells were cultured in vitro to investigate the antiallergic MSC-CMT mechanisms. Topical instillation of MSC-CMT significantly attenuated the clinical symptoms of short ragweed pollen-induced EAC, with a significant decrease in inflammatory cell frequency, nuclear factor κB p65 expression, and TNF-α and IL-4 production. In vitro MSC-CMT significantly inhibited the activation of MCs and B-cell IgE release and reduced histamine-induced vascular hyperpermeability. During EAC, MSC-CMT treatment also decreased IgE production, histamine release, enrichment and activation of MCs, and conjunctival vascular hyperpermeability. The MSC-CMT-mediated inhibition of B cells, MCs, and histamine and its antiallergic effects during EAC were abrogated when MSCs were pretreated with COX2 small interfering RNA. Our findings provide compelling evidence that MSC-CMT inhibits EAC through COX2-dependent multiple antiallergic mechanisms and support the use of MSC-CMT as a novel strategy for treating allergic conjunctivitis.


B-cell lymphomas with concurrent IGH-BCL2 and MYC rearrangements are aggressive neoplasms with clinical and pathologic features distinct from Burkitt lymphoma and diffuse large B-cell lymphoma.
B-cell lymphomas with concurrent IGH-BCL2 and MYC rearrangements, also known as "double-hit" lymphomas (DHL), are rare neoplasms characterized by highly aggressive clinical behavior, complex karyotypes, and a spectrum of pathologic features overlapping with Burkitt lymphoma (BL), diffuse large B-cell lymphoma (DLBCL) and B-lymphoblastic lymphoma/leukemia (B-LBL). The clinical and pathologic spectrum of this rare entity, including comparison to other high-grade B-cell neoplasms, has not been well defined. We conducted a retrospective analysis of clinical and pathologic features of 20 cases of DHL seen at our institution during a 5-year period. In addition, we carried out case-control comparisons of DHL with BL and International Prognostic Index (IPI)-matched DLBCL. The 11 men and 9 women had a median age of 63.5 years (range 32 to 91). Six patients had a history of grade 1 to 2 follicular lymphoma; review of the prior biopsy specimens in 2 of 5 cases revealed blastoid morphology. Eighteen patients had Ann Arbor stage 3 or 4 disease and all had elevated serum lactate dehydrogenase (LDH) levels at presentation. Extranodal disease was present in 17/20 (85%), bone marrow involvement in 10/17 (59%) and central nervous system (CNS) disease in 5/11 (45%). Nineteen patients were treated with combination chemotherapy, of whom 18 received rituximab and 14 received CNS-directed therapy. Fourteen patients (70%) died within 8 months of diagnosis. Median overall survival in the DHL group (4.5 mo) was inferior to both BL (P=0.002) and IPI-matched DLBCL (P=0.04) control patients. Twelve DHL cases (60%) were classified as B-cell lymphoma, unclassifiable, with features intermediate between DLBCL and BL, 7 cases (35%) as DLBCL, not otherwise specified, and 1 case as B-LBL. Distinguishing features from BL included expression of Bcl2 (P<0.0001), Mum1/IRF4 (P=0.006), Ki-67 <95% (P<0.0001), and absence of EBV-EBER (P=0.006). DHL commonly contained the t(8;22) rather than the t(8;14) seen in most BL controls (P=0.001), and exhibited a higher number of chromosomal aberrations (P=0.0009). DHL is a high-grade B-cell neoplasm with a poor prognosis, resistance to multiagent chemotherapy, and clinical and pathologic features distinct from other high-grade B-cell neoplasms. Familiarity with the morphologic and immunophenotypic spectrum of DHL is important in directing testing to detect concurrent IGH-BCL2 and MYC rearrangements when a karyotype is unavailable. The aggressive clinical behavior and combination of genetic abnormalities seen in these cases may warrant categorization as a separate entity in future classifications and call for novel therapeutic approaches.


Food web connections and the transmission cycles of Trypanosoma cruzi and Trypanosoma evansi (Kinetoplastida, Trypanosomatidae) in the Pantanal Region, Brazil.
We examined by parasitological tests (hemocultures and buffy coat) infection by Trypanosoma cruzi and T. evansi in blood samples from Leopardus pardalis, Cerdocyon thous and domestic dogs. Besides, 25 T. cruzi isolates previously derived from feral pigs and small wild mammals were here characterized by miniexon gene and demonstrated to be in the TcI genotype. Herein, we make an overall analysis of the transmission cycle of both trypanosome species in the light of the assemblage of data collected over the last seven years. The carnivore Nasua nasua was confirmed to play a major role in the transmission cycles of both T. cruzi and T. evansi since it was the species that had the higher prevalence and higher parasitemias by both flagellate species. In addition, our results show that both trypanosomatid species may be found throughout the Pantanal landscape, in all forest strata, as shown by the infection of carnivore, arboreal and terrestrial scansorial marsupial species in complex and seasonal transmission cycles. We propose that transmission of T. cruzi and T. evansi in the southern Pantanal region takes place via an intricate ecological trophic network involving generalist and specialist mammal species that are linked through a robust food-web connection.


Systemic corticosteroids in the treatment of acute exacerbations of chronic obstructive pulmonary disease.
The literature was reviewed to determine whether data support current treatment guideline recommendations regarding the use of systemic corticosteroids in the treatment of chronic obstructive pulmonary disease (COPD) exacerbations. Exacerbations of COPD are common and can be detrimental to both patient health and health care costs. Corticosteroids are recommended by consensus guidelines for patients during exacerbations of COPD. Although guidelines make very specific recommendations, clinical data are conflicting and inconsistent. A search of the English-language medical literature was performed, and all randomized, double-blind, placebo-controlled trials or meta-analyses that examined the use of systemic corticosteroids in COPD exacerbations were included. Trials that included nebulized corticosteroids were also included as long as they were compared to a systemic corticosteroid and a placebo. Recommendations regarding the use of systemic corticosteroids are not optimal or completely supported. Data support recommendations if patients are treated on an outpatient basis. However, hospitalized patients might also benefit from higher doses of systemic steroids initially, followed by an oral taper dose. In the treatment of COPD exacerbations, systemic corticosteroids improve airflow limitations, decrease treatment failure rates, decrease the risk of relapse, and may improve symptoms and decrease the length of hospital stay. Because of the risks of adverse effects, the lowest dose and shortest duration of corticosteroid therapy that will provide therapeutic benefit should be chosen. The literature suggests that hospitalized patients should benefit from a higher initial dosage of systemic corticosteroids than the 30-40 mg of i.v. or oral prednisolone for 7-10 days recommended in current guidelines.


Neurofibromatosis Type 1 with the Development of Pheochromocytoma and Breast Cancer.
A 40-year-old woman presented with a left adrenal incidentaloma. Based on the presence of café-au-lait spots, cutaneous neurofibroma, and family history, she was diagnosed with neurofibromatosis type 1 (NF1). Adrenal incidentaloma screening showed an elevated normetanephrine level; the left adrenal mass showed the uptake of I-123 meta-iodobenzylguanidine. She underwent left adrenalectomy, and pheochromocytoma was diagnosed. One year later, the results of a biopsy of a palpable mass in the left breast suggested invasive ductal carcinoma. The patient underwent neoadjuvant chemotherapy followed by left breast-conserving surgery. We herein report a rare case of an NF1 patient who developed both pheochromocytoma and breast cancer.


Radiological changes in the formation of Brodie's abscess by sequential magnetic resonance imaging: a case report.
Brodie's abscess is a relatively rare subacute form of osteomyelitis. Early diagnosis is challenging because of its insidious onset and vague symptoms. Magnetic resonance imaging is helpful in the diagnosis of Brodie's abscess; however, to date, no study has described the imaging findings of this disease in the early stage. Here, we present the case of a 14 year-old boy with Brodie's abscess in the proximal tibia. The lesion initially presented as a bone marrow edema in the proximal metaphysis of the left tibia on MRI and was misinterpreted as a bone bruise. Further radiological examination was performed 1 month later; this revealed the formation of an abscess cavity, which suggested Brodie's abscess. The patient was referred to our hospital and underwent curettage and debridement, which led to the definitive diagnosis of Brodie's abscess on histopathological findings and bacterial culture. On careful retrospective evaluation, the initial radiological findings suggested a microabscess on the metaphyseal side of the growth plate and bone marrow edema spreading from the lesion to the epiphysis. These radiological changes could be reliable evidence proving that the metaphyseal side of the growth plate is the origin of Brodie's abscess. Moreover, bone marrow edema with suspected microabscess in the metaphysis of the long bones can be the initial stage of the formation of Brodie's abscess and should be carefully followed up.


Cigarette smoke extract attenuates endothelium-dependent arteriolar dilatation in vivo.
The purpose of this study was to examine the effects of cigarette smoke extract on endothelium-dependent and endothelium-independent dilatation of arterioles in vivo. Using intravital microscopy, we measured diameter of arterioles contained within the microcirculation of the hamster cheek pouch during suffusion with acetylcholine and nitroglycerin, before and after treatment with cigarette smoke extract. Under control conditions, acetylcholine and nitroglycerin produced dose-related dilatation of cheek pouch arterioles. Superfusion of cigarette smoke extract (1.0%) did not alter baseline diameter of arterioles or vasodilatation in response to nitroglycerin but impaired dilatation of arterioles in response to acetylcholine. Next, we examined the possibility that impaired dilatation of cheek pouch arterioles in response to acetylcholine after exposure to cigarette smoke extract may be related to the release of substances produced via the cyclooxygenase pathway. In indomethacin-pretreated hamsters, acetylcholine produced similar vasodilatation before and after exposure to cigarette smoke extract. Thus these findings suggest that cigarette smoke extract impairs endothelium-dependent responses of cheek pouch arterioles. The mechanism of impaired responses of cheek pouch arterioles after exposure to cigarette smoke extract appears to be related to the release of substances produced via the cyclooxygenase pathway.


[The distribution of radiation-induced breaks in the chromosomes of irradiated subjects].
Distribution of radiation-induced breakpoints in chromosomes and its bands in persons recovered from acute radiation sickness and personnel from Chernobyl NPP were investigated using G-banding staining. The frequency of damaged bands and breakpoints in groups exposed to radiation was significantly higher as compared with the control group. It was shown that in exposed to radiation persons damage depends on its length. Most frequently damaged bands in the observed groups were determined. The G-negative bands and telomeres of chromosomes were more sensitive to radiation.


[Cholesteatoma, oto- and tympanosclerosis: their general mechanism of tissue destruction (author's transl)].
The connective tissue contains extracellular enzymes for the turnover of the ground substance. If this enzyme system gets out of control there is development of a complex metabolic disturbance of all tissue components. The lysosomal dysfunction is the basis of the general mechanism of tissue destruction in cholesteatoma, oto- and tympanosclerosis. This process seems most enhanced in cholesteatoma. Emphasis is placed on the role of extracellular lysosomes (= matrix vesicles).


Five-year survival results of subcutaneous low-dose immunotherapy with interleukin-2 alone in metastatic renal cell cancer patients.
After the discovery of its essential role in anticancer immunity, IL-2 cancer immunotherapy has shown that comparable results may be obtained with different schedules, including intravenous high-dose IL-2 as a bolus or as a 24-hour intravenous infusion or prolonged subcutaneous injection of low-dose IL-2 with or without IFN-alpha. This study shows the long-term results obtained in 92 metastatic renal cell cancer (RCC) patients with low-dose subcutaneous IL-2, which was given at 3 million IU twice/day for 5 days/week for 6 consecutive weeks. In nonprogressing patients, a second cycle was planned after a 21-day rest period, followed by maintenance therapy consisting of 5 days of treatment every month until disease progression. Complete response (CR) was achieved in only 2/92 (2%) patients, and partial response (PR) was observed in 19 patients (21%). Therefore, the response rate (CR + PR) was 21/92 (23%), with a median duration of response of 25 months. Stable disease (SD) occurred in 37 patients (40%), whereas the other 34 (37%) had a progressive disease (PD). The response rate was significantly higher in patients with a disease-free interval of >1 year than in those with a lower interval, in patients with a high performance status (PS) than in those with a low PS, and in patients with sites of disease other than the liver. A 5-year survival was obtained in 9/92 (9%) patients, and the percent of survival was significantly higher in patients with a response or SD than in those with PD. The treatment was well tolerated in all patients. This study confirms that low-dose subcutaneous IL-2 alone in an effective and well tolerated therapy of metastatic RCC, with results comparable to those described with more aggressive and toxic IL-2 schedules.


Outbreak of enterotoxigenic Escherichia coli O169 enteritis in schoolchildren associated with consumption of kimchi, Republic of Korea, 2012.
Enterotoxigenic Escherichia coli (ETEC) is now recognized as a common cause of foodborne outbreaks. This study aimed to describe the first ETEC O169 outbreak identified in Korea. In this outbreak, we identified 1642 cases from seven schools. Retrospective cohort studies were performed in two schools; and case-control studies were conducted in five schools. In two schools, radish kimchi was associated with illness; and in five other schools, radish or cabbage kimchi was found to have a higher risk among food items. Adjusted relative risk of kimchi was 5·87-7·21 in schools that underwent cohort studies; and adjusted odds ratio was 4·52-12·37 in schools that underwent case-control studies. ETEC O169 was isolated from 230 affected students, and was indistinguishable from the isolates detected from the kimchi product distributed by company X, a food company that produced and distributed kimchi to all seven schools. In this outbreak, we found that the risk of a kimchi-borne outbreak of ETEC O169 infection is present in Korea. We recommend continued monitoring regarding food safety in Korea, and strengthening surveillance regarding ETEC O169 infection through implementation of active laboratory surveillance to confirm its infection.


All hands on deck: early graduation of senior medical students in the COVID-19 pandemic.
This article was migrated. The article was marked as recommended. During the COVID-19 pandemic, early graduation of senior medical students simultaneously offers useful clinical experience in preparation for junior doctor posts, whilst helping address staffing shortages due to illness or self-isolation. Having recently graduated early from medical school, we offer our reflections on the obstacles and opportunities associated with working in an uncharted clinical environment. We are not the only ones on a steep learning curve at this time: this pandemic will challenge and provide learning for staff of all levels.


Triazole nucleoside derivatives bearing aryl functionalities on the nucleobases show antiviral and anticancer activity.
Synthetic nucleoside mimics are important candidates in the searing for antiviral and anticancer drugs. Ribavirin, the first antiviral nucleoside drug, is unique in its antiviral activity with mutilple modes of action, which are mainly due to its special triazole heterocycle as nucleobase. Additionally, introducing aromatic functionalities to the nucleobase is able to confer novel mechanisms of action for nucleoside mimics. With the aim to combine the special characteristics of unnatural triazole heterocycles with those of the appended aromatic groups on the nucleobases, novel 1,2,4-triazole nucleoside analogs bearing aromatic moieties were designed and developed. The present short review summarizes the molecular design, chemical synthesis and biological activity of these triazole nucleoside analogs. Indeed, the discovery of antiviral and anticancer activities shown by these triazole nucleosides as well as the new mechanism underlying the biological activity by one of the anticancer leads has validated the rationale for molecular design and impacted us to further explore the concept with the aim of developing structurally novel nucleoside drug candidates with new modes of action.


Experimental derivation of relative biological effectiveness of A-bomb neutrons in Hiroshima and Nagasaki and implications for risk assessment.
Epidemiological data on the health effects of A-bomb radiation in Hiroshima and Nagasaki provide the framework for setting limits for radiation risk and radiological protection. However, uncertainty remains in the equivalent dose, because it is generally believed that direct derivation of the relative biological effectiveness (RBE) of neutrons from the epidemiological data on the survivors is difficult. To solve this problem, an alternative approach has been taken. The RBE of polyenergetic neutrons was determined for chromosome aberration formation in human lymphocytes irradiated in vitro, compared with published data for tumor induction in experimental animals, and validated using epidemiological data from A-bomb survivors. The RBE of fission neutrons was dependent on dose but was independent of the energy spectrum. The same RBE regimen was observed for lymphocyte chromosome aberrations and tumors in mice and rats. Used as a weighting factor for A-bomb survivors, this RBE system was superior in eliminating the city difference in chromosome aberration frequencies and cancer mortality. The revision of the equivalent dose of A-bomb radiation using DS02 weighted by this RBE system reduces the cancer risk by a factor of 0.7 compared with the current estimates using DS86, with neutrons weighted by a constant RBE of 10.


Recurrent lower respiratory tract infections in children: a practical approach to diagnosis.
Many children are affected by recurrent lower respiratory tract infections (LRTIs), but the majority of them do not suffer from serious lung or extrapulmonary disease. The challenge for clinicians is to distinguish the recurrent RTIs with self-limiting or minor problems from those with underlying disease. The aim of this review is to describe a practical approach to children with recurrent LRTIs that limits unnecessary, expensive and time-consuming investigations. The children can be divided into three groups on the basis of their personal and family history and clinical findings: 1) otherwise healthy children who do not need further investigations; 2) those with risk factors for respiratory infections for whom a wait-and-see approach can be recommended; and 3) those in whom further investigations are mandatory. However, regardless of the origin of the recurrent LRTIs, it is important to remember that prevention by means of vaccines against respiratory pathogens (i.e. type b Haemophilus influenzae, pertussis, pneumococcal and influenza vaccines) can play a key role.


Validation of imatinib therapy failure score (IMTF) in chronic phase chronic myeloid leukemia in real life practice.
The outcome of chronic myeloid leukemia (CML) patients improved in the last decade. Clinical prognostic scoring systems aim to provide information about survival in the long-term, without determining from baseline the subset of patients who require a strictly monitoring because at increased risk of failure. Imatinib, the first-generation tyrosine kinase inhibitor (TKI), is still widely used as frontline treatment: recently, the imatinib therapy failure (IMTF) score was proposed to identify the failure free survival. Aim of our study was to validate this index in a large cohort of patients treated with imatinib.


Confined space manure storage and facilities safety assessment.
A mail survey of 1,200 farms across 16 states was conducted to identify the number, type, and size of manure storages per farm, as well as safety-related behaviors or actions related to entry into confined-space manure storage and handling facilities. Respondents provided data on 297 storage units and facilities, with approximately 75% reporting up to three storages per farm operation. Dimensions were provided for 254 manure pits: nearly 66% were less than or equal to 100 feet long, 75% were less than or equal to 40 feet wide, and 75% were less than or equal to 10 feet deep. Almost 14% of the reported storages were over 300 feet long, seven were wider than 100 feet, and 17 were more than 20 feet deep. Survey results suggest that most farm operations with confined-space manure storages do not follow best safety practices regarding their manure storages, including using gas detection equipment before entering a manure pit, using rescue lines when entering storages, or developing a written confined-space safety policy or plan. Survey results also suggest that few farmers post warning signs around their storages, post recommended ventilation times before entry, or conduct training for workers who enter confined-space manure storages. This article provides a benchmark against which the effectiveness of educational programs and design tools for confined-space manure pit ventilation systems and other confined-space manure pit safety interventions can be measured.


Associations of HLA-A, -B and -DRB1 types with oral diseases in Swiss adults.
Human leukocyte antigens (HLA) are crucial components of host defense against microbial challenge but the associations of HLA types with oral infectious diseases have not been studied in detail. This prospective cross-sectional study examined associations of HLA-A, -B and -DRB1 types with common oral diseases in a healthy Swiss adult population. 257 subjects (107 m, 150 f, mean age: 43.5 yr; range: 21-58 yr) with known HLA-A, -B and -DRB1 profiles and comprehensive medical records were included. A thorough anamnesis was followed by oral examinations including saliva flow measurements, the DMFT score for cariological status, complete periodontal status with plaque and bleeding indexes as well as assessment of mucosal alterations and temporomandibular dysfunction (TMD). Student's t-test and Pearson chi-square test were utilized to compare the oral diseases between HLA positive and negative subjects. Bonferroni correction for multiple comparisons was used and PBonf<0.05 was considered statistically significant. HLA types -B15 (PBonf = 0.002), -B51 (PBonf = 0.02) and -DRB1*12 (PBonf = 0.02) were associated with less periodontal disease manifestations. HLA-A32 had a positive association with TMD dysfunction (PBonf = 0.012). No other statistically significant associations were observed. In conclusion, HLA types may contribute to the development of oral diseases in generally healthy Caucasian adults.


Comparison of orbicularis oris muscle strength and endurance in young and elderly adults.
[Purpose] The purpose of this study was to compare the strength and endurance of the orbicularis oris muscle in healthy Korean (young vs. elderly adults). [Participants and Methods] A total of 60 participants (30 young adults and 30 elderly adults) were recruited. The Iowa Oral Performance Instrument was used to measure orbicularis oris muscle maximal strength and endurance. [Results] Elderly adults showed significantly lower orbicularis oris muscle strength and endurance than younger adults. [Conclusion] This study confirmed a significant age-related decrease in orbicularis oris muscle strength and endurance. The data collected will be useful as a basis for future on speech and swallowing therapy.


ATP-sensitive potassium currents reduce the PGE2-mediated enhancement of excitability in adult rat sensory neurons.
Behavioral studies have shown that the hyperalgesia arising from inflammatory agents, such as prostaglandin E(2) (PGE(2)), can be antagonized by activators of the ATP-sensitive potassium current (K(ATP)). This observation raises questions as to whether this suppression results from a direct action on sensory neurons and what are the cellular mechanisms giving rise to this inhibition. We found that small to medium diameter sensory neurons isolated from the L4-6 DRGs expressed the mRNAs for Kir6.1, Kir6.2, and SUR1. In perforated-patch clamp recordings from acutely dissociated sensory neurons from the young adult rat, exposure to 300 microM diazoxide, a K(ATP) channel agonist, significantly hyperpolarized the resting membrane potential, reduced the number of action potentials evoked by a ramp of depolarizing current, and increased the amplitude of inward K(ATP) currents evoked by the voltage ramp. Similar results were obtained with the protonophore FCCP, which is known to reduce the levels of intracellular ATP and lead to the activation of K(ATP). Only a subpopulation of sensory neurons was sensitive to diazoxide whereas other neurons were unaffected. Treatment with 1 microM PGE(2) significantly enhanced the excitability of these small to medium diameter capsaicin-sensitive sensory neurons; this enhancement was reversed by subsequent exposure to diazoxide in a subpopulation of neurons. Similar to diazoxide, exposure to 8-Br-cyclic GMP antagonized the PGE(2)-induced increase in excitability. The effects of 8-Br-cyclic GMP could be reversed by exposure to glibenclamide, an antagonist of K(ATP) channels. As with diazoxide, only a subpopulation of sensory neurons were affected by 8-Br-cyclic GMP. These results demonstrate that activation of K(ATP) can reverse the sensitization produced by PGE(2) and may be an important means to modulate the enhanced excitability that results from inflammatory or injury conditions.


Tangles and plaques in nondemented aging and "preclinical" Alzheimer's disease.
The distribution and density of neurofibrillary tangles and amyloid plaques was studied in a unique series of cases whose premortem cognitive status had been assessed with the Clinical Dementia Rating (CDR), including 39 nondemented cases (CDR = 0; age, 51-88 years), 15 very mildly demented cases (CDR = 0.5), and 8 severely demented (CDR = 3) cases. The initial formation of tangles and plaques in healthy aging appeared to be independent of each other. Tangles were found in all the nondemented cases, especially in hippocampal and parahippocampal areas; the average tangle concentration increased exponentially with age. In contrast, plaques were absent in some brains up to age 88, and the earliest plaque formation in other cases occurred in the neocortex, in patches of diffuse plaques. Widely distributed neuritic as well as diffuse plaques throughout neocortex and limbic structures characterized a further group of nondemented cases. In these cases there was also a substantial increase over other nondemented cases, both in the number of tangles and in the rate of increase in tangles with age, suggesting an interaction between amyloid and neurofibrillary change at this stage. Such cases closely resemble CDR = 0.5 cases, and it is proposed they represent "preclinical" Alzheimer's disease.


Idioventricular rhythm complicating acute myocardial infarction.
The incidence, natural history, prognosis, and electrocardiographic characteristics of idioventricular rhythm complicating acute myocardial infarction are described. It occurred as a transient arrhythmia nearly always within 24 hours of infarction in 61 (8%) of 737 patients, and was characterized by paroxysms of between 6 and 20 beats with widened bizarre QRS complexes at a rate of between 60 and 90 a minute. Most cases showed fusion beats and P waves dissociated from the QRS complexes, and in many cases idioventricular rhythm started during the slow phase of sinus arrhythmia. Though it usually occurred in patients with moderately severe transmural infarcts, the incidence of ventricular fibrillation and subsequent mortality was no greater than in patients with infarcts of equivalent severity who did not have idioventricular rhythm. It is concluded that this rhythm is a common and relatively benign arrhythmia complicating myocardial infarction, and that it should be distinguished from ventricular tachycardia.


Pharmacokinetics of an injectable modified-release 2-hydroxyflutamide formulation in the human prostate gland using a semiphysiologically based biopharmaceutical model.
The local distribution of 2-hydroxyflutamide (2-HOF) in prostate tissue after a single intraprostatic injection of a novel parenteral modified-release (MR) formulation in patients with localized prostate cancer was estimated using a semiphysiologically based biopharmaceutical model. Plasma concentration-time profiles for 2-HOF were acquired from a phase II study in 24 patients and the dissolution of the MR formulation was investigated in vitro. Human physiological values and the specific physicochemical properties of 2-HOF were obtained from the literature or calculated via established algorithms. A compartmental modeling approach was adopted for tissue and blood in the prostate gland, where the compartments were modeled as a series of concentric spherical shells contouring the centrally positioned depot formulation. Discrete fluid connections between the blood compartments were described by the representative flow of blood, whereas the mass transport of drug from tissue to tissue and tissue to blood was described by a one-dimensional diffusion approximation. An empirical dissolution approach was adopted for the release of 2-HOF from the formulation. The model adequately described the plasma concentration-time profiles of 2-HOF. Predictive simulations indicated that the local tissue concentration of 2-HOF within a distance of 5 mm from the depot formulation was approximately 40 times higher than that of unbound 2-HOF in plasma. The simulations also indicated that spreading the formulation throughout the prostate gland would expose more of the gland and increase the overall release rate of 2-HOF from the given dose. The increased release rate would initially increase the tissue and plasma concentrations but would also reduce the terminal half-life of 2-HOF in plasma. Finally, an in vitro-in vivo correlation of the release of 2-HOF from the parenteral MR formulation was established. This study shows that intraprostatic 2-HOF concentrations are significantly higher than systemic plasma concentrations and that increased distribution of 2-HOF throughout the gland, using strategic imaging-guided administration, is possible. This novel parenteral MR formulation, thus, facilitates good pharmacological effect while minimizing the risk of side effects.


The Frequency of Chromosomal Euploidy Among 3PN Embryos.
The evaluation of embryo morphology is one of the most important parameters used to evaluate developmental timing, also providing an indication of chromosomal failure or degeneration. The first step in the evaluation of a fertilization event is determining the number and shape of the pronuclei (PN). Normally fertilized eggs possess two even PN. However, some embryos which develop from abnormally fertilized zygotes may be tri-pronuclear zygotes (3PN). Thirty embryos were collected from 12 women who underwent in vitro fertilization (IVF) at Dr. Cipto Mangunkusumo General Hospital in Jakarta, Indonesia. Embryos were cultured until the blastocyst stage on days 5-6. The blastomere biopsy was performed by piercing the zona pellucida with a laser under a microscope. Chromosomal numerical abnormalities were analyzed using Next Generation Sequencing (NGS). Among the 30 embryos with 3PN zygotes, 33.3% had a normal chromosomal array, with 22 pairs of autosomes and 2 pairs of sex chromosomes. While the rest of sample population detected as abnormal chromosome (66.7%), with the highest percentage of abnormality was triploidy 43.3%, followed by mosaicism 13.4% and aneuploidy 10%. This was a preliminary study revealed not all morphologically 3PN embryos are genetically abnormal.


Structural coordination between active sites of a CRISPR reverse transcriptase-integrase complex.
CRISPR-Cas systems provide adaptive immunity in bacteria and archaea, beginning with integration of foreign sequences into the host CRISPR genomic locus and followed by transcription and maturation of CRISPR RNAs (crRNAs). In some CRISPR systems, a reverse transcriptase (RT) fusion to the Cas1 integrase and Cas6 maturase creates a single protein that enables concerted sequence integration and crRNA production. To elucidate how the RT-integrase organizes distinct enzymatic activities, we present the cryo-EM structure of a Cas6-RT-Cas1-Cas2 CRISPR integrase complex. The structure reveals a heterohexamer in which the RT directly contacts the integrase and maturase domains, suggesting functional coordination between all three active sites. Together with biochemical experiments, our data support a model of sequential enzymatic activities that enable CRISPR sequence acquisition from RNA and DNA substrates. These findings highlight an expanded capacity of some CRISPR systems to acquire diverse sequences that direct CRISPR-mediated interference.


Computing geometric properties of images represented by linear quadtrees.
The region quadtree is a hierarchical data structure that finds use in applications such as image processing, computer graphics, pattern recognition, robotics, and cartography. In order to save space, a number of pointerless quadtree representations (termed linear quadtrees) have been proposed. One representation maintains the nodes in a list ordered according to a preorder traversal of the quadtree. Using such an image representation and a graph definition of a quadtree, a general algorithm to compute geometric image properties such as the perimeter, the Euler number, and the connected components of an image is developed and analyzed. The algorithm differs from the conventional approaches to images represented by quadtrees in that it does not make use of neighbor finding methods that require the location of a nearest common ancestor. Instead, it makes use of a staircase-like data structure to represent the blocks that have been already processed. The worst-case execution time of the algorithm, when used to compute the perimeter, is proportional to the number of leaf nodes in the quadtree, which is optimal. For an image of size 2n × 2n, the perimeter algorithm requires only four arrays of 2n positions each for working storage. This makes it well suited to processing linear quadtrees residing in secondary storage. Implementation experience has confirmed its superiority to existing approaches to computing geometric properties for images represented by quadtrees.


The cellular mechanisms associated with the anesthetic and neuroprotective properties of xenon: a systematic review of the preclinical literature.
Xenon exhibits significant neuroprotection against a wide range of neurological insults in animal models. However, clinical evidence that xenon improves outcomes in human studies of neurological injury remains elusive. Previous reviews of xenon's method of action have not been performed in a systematic manner. The aim of this review is to provide a comprehensive summary of the evidence underlying the cellular interactions responsible for two phenomena associated with xenon administration: anesthesia and neuroprotection. A systematic review of the preclinical literature was carried out according to the PRISMA guidelines and a review protocol was registered with PROSPERO. The review included both in vitro models of the central nervous system and mammalian in vivo studies. The search was performed on 27th May 2022 in the following databases: Ovid Medline, Ovid Embase, Ovid Emcare, APA PsycInfo, and Web of Science. A risk of bias assessment was performed utilizing the Office of Health Assessment and Translation tool. Given the heterogeneity of the outcome data, a narrative synthesis was performed. The review identified 69 articles describing 638 individual experiments in which a hypothesis was tested regarding the interaction of xenon with cellular targets including: membrane bound proteins, intracellular signaling cascades and transcription factors. Xenon has both common and subtype specific interactions with ionotropic glutamate receptors. Xenon also influences the release of inhibitory neurotransmitters and influences multiple other ligand gated and non-ligand gated membrane bound proteins. The review identified several intracellular signaling pathways and gene transcription factors that are influenced by xenon administration and might contribute to anesthesia and neuroprotection. The nature of xenon NMDA receptor antagonism, and its range of additional cellular targets, distinguishes it from other NMDA antagonists such as ketamine and nitrous oxide. This is reflected in the distinct behavioral and electrophysiological characteristics of xenon. Xenon influences multiple overlapping cellular processes, both at the cell membrane and within the cell, that promote cell survival. It is hoped that identification of the underlying cellular targets of xenon might aid the development of potential therapeutics for neurological injury and improve the clinical utilization of xenon. https://www.crd.york.ac.uk/prospero/, identifier: 336871.


Three-dimensional reconstruction of icosahedral particles from single micrographs in real time at the microscope.
Single particle analysis is a valuable tool in cryo-electron microscopy for determining the structure of biological complexes. However, the conformational state and the preparation of the sample are factors that play a critical role in the ultimate attainable resolution. In some cases extensive analysis at the microscope of a sample under different conditions is required to derive the optimal acquisition conditions. Currently this analysis is limited to raw micrographs, thus conveying only limited information on the structure of the complex. We are developing a computing system that generates a three-dimensional reconstruction from a single micrograph acquired under cryogenic and low dose conditions, and containing particles with icosahedral symmetry. The system provides the microscopist with immediate structural information from a sample while it is in the microscope and during the preliminary acquisition stage. The system is designed to run without user intervention on a multi-processor computing resource and integrates all the processing steps required for the analysis. Tests performed on experimental data sets show that the probability of obtaining a reliable reconstruction from one micrograph is primarily determined by the quality of the sample, with success rates close to 100% when sample conditions are optimal, and decreasing to about 60% when conditions are sub-optimal. The time required to generate a reconstruction depends significantly on the diameter of the particles, and in most instances takes about 1min. The proposed approach can provide valuable three-dimensional information, albeit at low resolution, on conformational states, epitope binding, and stoichiometry of icosahedral multi-protein complexes.


Ambulatory care pavilion takes its place out front by solving multiple needs.
In sum, this structure exemplifies the fact that high-tech tertiary care medical centers can be user-friendly to the ambulatory health care consumer by serving their routine needs conveniently and efficiently. Says Gerald Miller, president of Crozer-Chester: "The ambulatory care pavilion has enabled Crozer to successfully and efficiently merge physicians' offices with institutional-based services and inpatient services. We are pleased with how the pavilion positions our medical center for the next century.


Soluble Receptor for Advanced Glycation End Products (sRAGE) Is a Sensitive Biomarker in Human Pulmonary Arterial Hypertension.
Pulmonary arterial hypertension (PAH) is a progressive condition with an unmet need for early diagnosis, better monitoring, and risk stratification. The receptor for advanced glycation end products (RAGE) is activated in response to hypoxia and vascular injury, and is associated with inflammation, cell proliferation and migration in PAH. For the adult cohort, we recruited 120 patients with PAH, 83 with idiopathic PAH (IPAH) and 37 with connective tissue disease-associated PAH (CTD-PAH), and 48 controls, and determined potential plasma biomarkers by enzyme-linked immunoassay. The established heart failure marker NTproBNP and IL-6 plasma levels were several-fold higher in both adult IPAH and CTD-PAH patients versus controls. Plasma soluble RAGE (sRAGE) was elevated in IPAH patients (3044 ± 215.2 pg/mL) and was even higher in CTD-PAH patients (3332 ± 321.6 pg/mL) versus controls (1766 ± 121.9 pg/mL; p < 0.01). All three markers were increased in WHO functional class II+III PAH versus controls (p < 0.001). Receiver-operating characteristic analysis revealed that sRAGE has diagnostic accuracy comparable to prognostic NTproBNP, and even outperforms NTproBNP in the distinction of PAH FC I from controls. Lung tissue RAGE expression was increased in IPAH versus controls (mRNA) and was located predominantly in the PA intima, media, and inflammatory cells in the perivascular space (immunohistochemistry). In the pediatric cohort, plasma sRAGE concentrations were higher than in adults, but were similar in PH (n = 10) and non-PH controls (n = 10). Taken together, in the largest adult sRAGE PAH study to date, we identify plasma sRAGE as a sensitive and accurate PAH biomarker with better performance than NTproBNP in the distinction of mild PAH from controls.


Analysis of South American climate and teleconnection indices.
Oceanic heat anomalies affect climate in remote regions through the atmospheric cycle. South America (SA) was the first region found associated with EI Niño, which affects the fishery, agriculture, forestry, and livestock industry of SA. As approximately 60% of the total water is used for agriculture, climate changes in SA caused by ocean anomalies have led to the variability of available water, especially for irrigation water. Where the precipitation is low and/or the temperature is high, the availability and quality of water resources are under pressure. For instance, droughts associated with La Niña severely limited water supply and irrigation requirements between 25°S - 40°S in west-central Argentina and central Chile. In order to study the relationship between ocean variability and the climate of SA, 19 teleconnection indices (TI) related to Ocean abnormity are considered. The 19 indices are: the sea surface temperature (SST) and their anomaly in 4 Niño regions (SST1 + 2, SST3, SST3.4, SST4, ANOM1 + 2, ANOM3, ANOM3.4, ANOM4), Southern Oscillation Index (SOI), Oceanic Niño Index (ONI), Outgoing Longwave Radiation (OLR), Arctic Oscillation (AO), North Atlantic Oscillation (NAO), Pacific Decadal Oscillation (PDO), Pacific-North America (PNA), Atlantic Multi-decadal Oscillation (AMO), West and East of Indian Ocean Dipole (IODW, IODE), and the difference between IODW and IODE (IODd). High-resolution gridded climate data (1982-2016) from the Global Precipitation Climatology Centre (GPCC), the Climate Prediction Center (CPC), and the National Centers for Environmental Prediction (NCEP) are applied for correlation analyses. The results show that the 89.4% area of South American climate has a significant correlation with the SST in Niño region 1 + 2, the mean correlation coefficient is 0.55 for NCEP precipitation and 0.54 for CPC temperature. The lag duration for the remote correlation is around 2-3 months. It is the first attempt to analyze the correlation relationship based on 19 TIs, which can provide comprehensive insight into the climate of SA at a high-resolution scale. These findings are helpful for identifying the sensitive factors that affect climate in SA, for projecting the climate variables of SA, and for managing the irrigation water resources of SA.


Refractive collimation beam shaper design and sensitivity analysis using a free-form profile construction method.
A refractive laser beam shaper comprising two free-form profiles is presented. The profiles are designed using a free-form profile construction method such that each incident ray is directed in a certain user-specified direction or to a particular point on the target surface so as to achieve the required illumination distribution of the output beam. The validity of the proposed design method is demonstrated by means of ZEMAX simulations. The method is mathematically straightforward and easily implemented in computer code. It thus provides a convenient tool for the design and sensitivity analysis of laser beam shapers and similar optical components.


Beyond the professional fee, Part 1: Basic legal issues for physicians investing in the technical side of healthcare services.
As reimbursement for professional services shrinks in comparison with physicians' educational debt and practice expenses, physicians are looking for additional sources of revenue to supplement their practice income. A natural option to consider is investment in healthcare entities or equipment in order to share in the technical fees generated by such entities or equipment.


Virulence and local adaptation of a horizontally transmitted parasite.
Parasites are thought to maximize the number of successfully transmitted offspring by trading off propagule production against host survival. In a horizontally transmitted microparasitic disease in Daphnia, a planktonic crustacean, increasing geographic distance between host and parasite origin was found to be correlated with a decrease in spore production and virulence. This finding indicates local adaptation of the parasite, but contradicts the hypothesis that long-standing coevolved parasites are less virulent than novel parasites. Virulence can be explained as the consequence of balancing the positive genetic correlation between host mortality and strain-specific spore production.


A Randomized Trial Comparing Freely Moving and Zonal Instruction of Veterinary Surgical Skills Using Ovariohysterectomy Models.
Timely, specific feedback is the most important feature of simulation-based training, but providing adequate instructor supervision is challenging. Students' (n = 76) surgical skills were assessed after training using either the traditional (T) method of large-group teaching by multiple instructors or the alternative method of one instructor assigned (A) to a defined group of students. Instructors rotated to a different group of students for each laboratory session. The instructor-to-student ratio and environment remained identical. No differences were found in raw assessment scores or the number of students requiring remediation, suggesting that students learned in this environment whether they received feedback from one instructor or multiple. Students had no preference between the methods, though 88% of the instructors preferred the assigned method, because they perceived an increased ability to teach and observe individual students. There was no difference in the number of students identified as at-risk of remediation between groups. When both groups were considered together, students identified as at-risk were more likely (40% vs. 10%) to require post-assessment remediation. However, only 22% of students requiring remediation had been identified as at-risk, and A-group instructors were more accurate than T-group instructors at identifying at-risk students. These results suggest that students accept either instructional method, but most instructors prefer to be assigned to a small group of students. Surgical skills were learned similarly well by students in both groups, although assigned instructors were more accurate at identifying at-risk students, which could prove beneficial if early intervention measures can be offered.


[Tissue Intrinsic Fluorescence Spectrum Recovering Based on Diffusion Theory].
Tissue intrinsic fluorescence spectrum refers to the fluorescence that is not impaired by tissue absorption and scattering which has the ability to reflect tissue biochemical properties. In order to reduce the influence of tissue absorption and scattering properties on tissue fluorescence spectrum, and then recover tissue intrinsic fluorescence spectrum, a tissue spectrum detection system based on fiber-optic probe was developed for the measurement of tissue fluorescence spectrum and diffusion reflectance spectrum at the same place. On the other hand, diffusion theory was introduced to extract the tissue physiological parameters from the measurement tissue diffusion reflectance spectrum, which included blood volume fraction, oxyhemoglobin saturation, melanin content, reduce scattering coefficient at 500 nm and the ratio of rayleigh scattering and the total scattering. Then tissue optical parameters in visible wavelengths were calculated. According to the tissue optical parameters and measured tissue diffusion spectrum, the intrinsic fluorescence spectrum was recovered from the measured fluorescence. Based on this, clinical trials were conducted to measure human skin fluorescence spectrum and diffusion reflectance spectrum, and then to recover skin intrinsic fluorescence spectrum. Finally, the accumulation of Advanced Glycation End products (AGE) in human skin was evaluated and the probability of diabetes mellitus was predicted. The result shows that the sensitivity and specificity were 69% and 0.75% respectively, when the measured fluorescent was used to screening diabetes mellitus. At the same specificity, the sensitivity was 90% when the recovered intrinsic fluorescence was employed to screening diabetes mellitus.


Primary or secondary antifungal prophylaxis in patients with hematological maligancies: efficacy and damage.
Patients with hematological malignancies often develop febrile neutropenia (FN) as a complication of cancer chemotherapy. Primary or secondary antifungal prophylaxis is recommended for patients with hematological malignancies to reduce the risk of invasive fungal infection (IFI). This study retrospectively evaluated the efficacy and potential harm of administration of primary and secondary antifungal prophylaxis to patients with hematological malignancies at one hospital. All patients with hematological malignancies older than 14 years of age who had experienced at least one FN attack during chemotherapy while being treated at one hospital between November 2010 and November 2012 were retrospectively evaluated. A total of 282 FN episodes in 126 consecutive patients were examined during a 2-year study period. The mean patient age was 51.73±14.4 years (range: 17-82 years), and 66 patients were male. Primary prophylaxis with posaconazole was administered to 13 patients and systemic antifungal treatment under induction or consolidation chemotherapy to seven patients. Of 26 patients who received secondary antifungal prophylaxis with either oral voriconazole (n=17) or posaconazole (n=6) during 46 FN episodes, systemic antifungal therapy was administered in 16 of 38 episodes and three of eight episodes, respectively. Secondary antifungal prophylaxis with caspofungin was found effective in treating six FN episodes in three patients who had experienced at least two persistent candidemia attacks. The mortality rates associated with IFI were 9% in the first year, 2% in the second year, and 6% overall. The mortality rates associated with candidemia were 33% in the first year, 22% in the second year, and 27% overall. Primary antifungal prophylaxis should be administered to selected patients on the basis of consideration of efficacy, cost, and potential harm. Use of secondary prophylaxis may reduce systemic antifungal use and IFI frequency but may increase risk of colonization and infection with azole-resistant fungal strains.


A microwave heating technique for the hyperthermic treatment of tumours in the eye, especially retinoblastoma.
A special microwave stripline applicator (2450 MHz) has been designed for the hyperthermic treatment of eye tumours, especially retinoblastoma. The measured microwave absorbed power distribution was put into a thermal finite difference computer model of the human and rabbit eye to calculate the resulting temperature distributions. The temperature distributions have been experimentally verified on rabbit eyes, and satisfactory temperature distributions can be obtained. The anterior part as well as the posterior part of the eye can be heated uniformly to temperatures in the range of 37-44 degrees C. The complete hyperthermia system is described.


Pt nanocluster size effects in the hydrogen evolution reaction: approaching the theoretical maximum activity.
Hydrogen production from electrocatalytic water splitting in electrolyzers is a key process to store excess electric energy produced from intermittent renewable energy sources. For proton exchange membrane (PEM) electrolyzers, carbon supported platinum particles exhibit the highest rates for the hydrogen evolution reaction (HER); however, high Pt costs limit the wide spread use of this technology. By employing a graphene layer grown on a Ru(0001) single crystal as a template for Pt nanocluster (NC) growth, we studied the dependence of the HER activity on the NC size using NCs of different sizes. We provide clear quantitative experimental evidence for a volcano-like relationship between the HER activity and the NC size which has been missing so far. For Pt NCs with very low sizes below 2 nm, we found stunningly improved exchange HER current densities. The highest exchange current density was observed for Pt NCs with an average size of ca. 38 atoms. These Pt38 NCs do not only surpass the Pt-mass-specific activity of commercial Pt electrode materials by well above three orders of magnitude, also their exchange current density is located close to the maximum exchange current density for the HER predicted theoretically for transition metal surfaces. The present work provides a strong stimulus for future research towards technically feasible Pt NC catalysts with cluster sizes in the range of few tens of Pt atoms.


Slowing the progression of vascular calcification in hemodialysis.
Hyperphosphatemia and secondary hyperparathyroidism are common complications of ESRD (chronic kidney disease stage 5) that, when untreated, may result in increased morbidity and mortality. Hyperphosphatemia and hypercalcemia have been associated with increased coronary artery calcification. Achieving control of serum phosphorus without increasing serum calcium is an important goal for patients with ESRD. Although calcium-based phosphate binders effectively reduce serum phosphorus and parathyroid hormone concentrations, these agents can lead to hypercalcemia and have been associated with increased vascular calcification. The phosphorus binder sevelamer was developed to overcome the limitations associated with the usual management of hyperphosphatemia and secondary hyperparathyroidism (i.e., mineral salts). Sevelamer, a nonabsorbable hydrogel, is as efficacious as calcium-based phosphate binders for reducing serum phosphorus but does not cause hypercalcemia or other adverse metabolic effects. Sevelamer also exhibits beneficial effects on lipids, consistently and significantly decreasing LDL cholesterol and increasing HDL cholesterol in most studies. In a head-to-head randomized clinical trial, sevelamer and calcium-based binders achieved similarly excellent phosphorus control, but the use of calcium-based binders led to significantly higher serum calcium concentrations and an increased incidence of hypercalcemia and unintended suppression of parathyroid hormone. Treatment with calcium-based binders also led to the progression of coronary artery and aortic calcification, whereas sevelamer attenuated or arrested progression. Strategies that use oral calcium and vitamin D in patients with ESRD should be reexamined, and the potential advantages of sevelamer should be considered when selecting a primary agent to reduce serum phosphorus in hemodialysis patients.


Acceptance threshold theory can explain occurrence of homosexual behaviour.
Same-sex sexual behaviour (SSB) has been documented in a wide range of animals, but its evolutionary causes are not well understood. Here, we investigated SSB in the light of Reeve's acceptance threshold theory. When recognition is not error-proof, the acceptance threshold used by males to recognize potential mating partners should be flexibly adjusted to maximize the fitness pay-off between the costs of erroneously accepting males and the benefits of accepting females. By manipulating male burying beetles' search time for females and their reproductive potential, we influenced their perceived costs of making an acceptance or rejection error. As predicted, when the costs of rejecting females increased, males exhibited more permissive discrimination decisions and showed high levels of SSB; when the costs of accepting males increased, males were more restrictive and showed low levels of SSB. Our results support the idea that in animal species, in which the recognition cues of females and males overlap to a certain degree, SSB is a consequence of an adaptive discrimination strategy to avoid the costs of making rejection errors.


Exploring Overweight Risk Trajectories During Childhood and Their Associations With Elevated Blood Pressure at Late Adolescence: a Retrospective Cohort Study.
Overweight during childhood is significantly associated with higher risk of high blood pressure (HBP) in later life. However, recognition of critical intervention period is limited. We aimed to analyze the changes in overweight risk during school-age and its relationship with HBP. Seventeen thousand eight hundred sixteen school-aged children (53.9% boys) with a mean follow-up time of 8.2 years were involved. Children's overweight was defined as body mass index Z score ≥1 and was fitted with a group-based trajectory model. The 4 trajectories were labeled as constant low, high decreasing, low rising, and constant high according to the change of overweight risk during follow-up. Population-averaged logit model and log-binomial regression models were used to analyze HBP risk. Cox proportional hazard model was used to analyze the HBP incidence among distinct groups. Children with higher overweight risk generally had higher HBP risk during follow-up. However, the HBP risk in low rising group was low at baseline and increased to 3.14 (95% CI, 2.54-3.88; P<0.001) for boys and 3.23 (95% CI, 2.08-5.01; P=0.004) for girls at end point, which were comparable to the relative risk in the constant high group (4.60 [95% CI, 4.02-5.27] for boys and 5.28 [95% CI, 3.94-7.07] for girls). Findings of this study provide insights on patterns of overweight risks during childhood. Children with transition from normal to overweight during teen years would be high-risk in HBP incidence.


A portable high-field pulsed-magnet system for single-crystal x-ray scattering studies.
We present a portable pulsed-magnet system for x-ray studies of materials in high magnetic fields (up to 30 T). The apparatus consists of a split-pair of minicoils cooled on a closed-cycle cryostat, which is used for x-ray diffraction studies with applied field normal to the scattering plane. A second independent closed-cycle cryostat is used for cooling the sample to near liquid helium temperatures. Pulsed magnetic fields (approximately 1 ms in total duration) are generated by discharging a configurable capacitor bank into the magnet coils. Time-resolved scattering data are collected using a combination of a fast single-photon counting detector, a multichannel scaler, and a high-resolution digital storage oscilloscope. The capabilities of this instrument are used to study a geometrically frustrated system revealing strong magnetostrictive effects in the spin-liquid state.


The potential impact of native Australian trypanosome infections on the health of koalas (Phascolarctos cinereus).
Whole blood collected from koalas admitted to the Australian Zoo Wildlife Hospital (AZWH), Beerwah, QLd, Australia, during late 2006-2009 was tested using trypanosome species-specific 18S rDNA PCRs designed to amplify DNA from Trypanosoma irwini, T. gilletti and T. copemani. Clinical records for each koala sampled were reviewed and age, sex, blood packed cell volume (PCV), body condition, signs of illness, blood loss, trauma, chlamydiosis, bone marrow disease, koala AIDS and hospital admission outcome ('survival'/ 'non-survival') were correlated with PCR results. Overall 73.8% (439/595) of the koalas were infected with at least 1 species of trypanosome. Trypanosoma irwini was detected in 423/595 (71.1%), T. gilletti in 128/595 (21.5%) and T. copemani in 26/595 (4.4%) of koalas. Mixed infections were detected in 125/595 (21%) with co-infections of T. irwini and T. gilletti (101/595, 17%) being most common. There was a statistical association between infection with T. gilletti with lower PCV values and body condition scores in koalas with signs of chlamydiosis, bone marrow disease or koala AIDS. No association between T. gilletti infection and any indicator of health was observed in koalas without signs of concurrent disease. This raises the possibility that T. gilletti may be potentiating other disease syndromes affecting koalas.


Synthesis of DNA in isolated pancreatic islets maintained in tissue culture.
The DNA synthesis of isolated pancreatic islets has been determined after culture in vitro of the islets for 5 days in media containing either various glucose concentrations or tolbutamide or cortisone. Only glucose stimulated the replication of DNA, while cortisone was inhibitory and tolbutamide without effect. It is concluded that glucose by itself stimulates the mitotic rate of the islet cells and that the corresponding effect of steroids in vivo may be mediated through the ensuing hyperglycemia.


Right ventricular-pulmonary artery coupling in chronic thromboembolic pulmonary hypertension.
Chronic thromboembolic pulmonary hypertension occurs in a proportion of patients with prior acute pulmonary embolism and is characterised by breathlessness, persistently raised pulmonary pressures and right heart failure. Surgical pulmonary endarterectomy (PEA) offers significant prognostic and symptomatic benefits for patients with proximal disease distribution. For those with inoperable disease, management options include balloon pulmonary angioplasty (BPA) and medical therapy. Current clinical practice relies on the evaluation of pulmonary haemodynamics to assess disease severity, timing of and response to treatment. However, pulmonary haemodynamics correlate poorly with patient symptoms, which are influenced by right ventricular tolerance of the increased afterload. How best to manage symptomatic patients with chronic thromboembolic pulmonary disease (CTEPD) in the absence of pulmonary hypertension is not resolved.Right ventricular-pulmonary artery coupling (RV-PAC) describes the energy transfer within the whole cardiopulmonary unit. Thus, it can identify the earliest signs of decompensation even before pulmonary hypertension is overt. Invasive measurement of coupling using pressure volume loop technology is well established in research settings. The development of efficient and less invasive measurement methods has revived interest in coupling as a viable clinical tool. Significant improvement in RV-PAC has been demonstrated after both PEA and BPA. Further studies are required to understand its clinical utility and prognostic value, in particular, its potential to guide management in patients with CTEPD. Finally, given the reported differences in coupling between sexes in pulmonary arterial hypertension, further work is required to understand the applicability of proposed thresholds for decoupling in therapeutic decision making.


Posterior reversible encephalopathy syndrome: clinical and radiological manifestations, pathophysiology, and outstanding questions.
Almost two decades have elapsed since posterior reversible encephalopathy syndrome (PRES) was described in an influential case series. This usually reversible clinical syndrome is becoming increasingly recognised, in large part because of improved and more readily available brain imaging. Although the pathophysiological changes underlying PRES are not fully understood, endothelial dysfunction is a key factor. A diagnosis of PRES should be considered in the setting of acute neurological symptoms in patients with renal failure, blood pressure fluctuations, use of cytotoxic drugs, autoimmune disorders, or eclampsia. Characteristic radiographic findings include bilateral regions of subcortical vasogenic oedema that resolve within days or weeks. The presence of haemorrhage, restricted diffusion, contrast enhancement, and vasoconstriction are all compatible with a diagnosis. In most cases, PRES resolves spontaneously and patients show both clinical and radiological improvements. The range of symptoms that can comprise the syndrome might be broader than usually thought. In its mild form, this disorder might cause only one clinical symptom (headache or seizure) and radiographically might show few areas of vasogenic oedema or even normal brain imaging in some rare cases. In severe forms, PRES might cause substantial morbidity and even mortality, most often as a result of acute haemorrhage or massive posterior fossa oedema causing obstructive hydrocephalus or brainstem compression.


An Empirical Test of the Three-Step Theory of Suicide in U.K. University Students.
The purpose of this study was to further our understanding of how individuals move down the pathway from first thinking about suicide to ultimately attempting to take their own lives by empirically testing the Three-Step Theory (3ST) in a sample of university students (n = 665). Results largely support the theory's central propositions. First, an interactive model of pain and hopelessness accounted for substantial variance in suicidal desire. This result replicated in both men and women, and across age groups (i.e., below 35 and at or above 35). Also, as predicted, connectedness was protective against ideation in those high on both pain and hopelessness. However, contrary to our prediction, connectedness was similarly protective among everyone else. Finally, suicide capacity predicted suicide attempt history over and above current and lifetime suicide ideation. These findings provide further support to the 3ST.


Menstrual health and the metabolic syndrome in adolescents.
The metabolic syndrome, a constellation of interrelated risk factors for cardiovascular disease and type 2 diabetes mellitus, has become a major public health concern against the backdrop of increasing rates of obesity. Insulin resistance plays a pivotal role as the underlying pathophysiological linchpin of the various components of the syndrome. The metabolic syndrome is well recognized in adults, and there is convincing evidence that it starts in childhood, with progressive clustering of the various components over time and tracking through adulthood. Adult women and adolescents with polycystic ovary syndrome (PCOS) have higher prevalence rates of the metabolic syndrome compared with the general population. Several anthropometric (obesity, particularly abdominal obesity), metabolic (insulin resistance/hyperinsulinemia, dyslipidemia) and hormonal (low IGFBP1, IGFBP2 and low sex hormone binding globulin) features of adolescents with PCOS are also features of the metabolic syndrome. Insulin resistance, believed to be a key pathogenic factor in both PCOS and the metabolic syndrome, may be the thread that links the two conditions. Menstrual health in adolescents could be viewed as yet another component in the evaluation of the metabolic syndrome. Careful assessment of menstrual history and appropriate laboratory work-up could reveal the presence of PCOS in obese at-risk adolescent girls with a family history of the metabolic syndrome.


Histochemical changes in the testes of lead induced experimental rats.
The experiments were performed on mature male rats divided in five groups, one control and four experimental in which the animals received 1 mg, 2 mg, 4 mg and 6 mg/kg body weight lead acetate intraperitoneally respectively, over a period of 30 days. ALA-D and lead was estimated in the blood by the use of atomic absorption spectrophotometer and ATP-ase, AMP-ase, Alk-ase were histochemically localized. Significant increase in blood and testis of lead levels along with decrease of ALA-D levels were observed. Changes in the testicular tissue were encountered. Other details concerned with the damage of the testicular tissue are discussed.


Antisense yycG Regulation of Antibiotic Sensitivity of Methicillin-Resistant Staphylococcus aureus in Chronic Osteomyelitis.
Background: Methicillin-resistant Staphylococcus aureus (MRSA) is an urgent medical problem in osteomyelitis. The YycFG two-component regulatory system (TCS) allows bacteria to adapt rapidly to physical, chemical, and biological stresses. The recombinant plasmid shuttle vector was used to overexpress an antisense RNA (asRNA) to inhibit target gene expression by sequence-specific double-stranded RNA complex degradation. In the current study, antisense yycG RNA (ASyycG)-overexpression MRSA clinical isolates were constructed. Methods: Bacterial growth was monitored, and biofilm biomass was determined by crystal violet microtiter assay. Quantitative reverse transcription polymerase chain reaction analysis was used to identify expression of yycF/G/H and icaA/D in MRSA and ASyycG strains. The expression of YycG protein was quantified by Western blot assays. The antibiotic resistance of ASyycG strains was compared with that of the MRSA strains. Results: The ASyycG strains showed a decrease in growth rate compared with the MRSA strains. Of note, overexpression of ASyycG led to a reduction in biofilm formation and adhesion force. ASyycG strains had decreased expressions of the yycF/G/H and icaA/D. Furthermore, Western blot data showed that expression of the YycG protein decreased by 40% in ASyycG strains compared with MRSA strains. In addition, the effect of yycG asRNA improved the susceptibility of ASyycG strains to cefoxitin. Conclusions: The ASyycG strains inhibited biofilm organization and increased antibiotic sensitivity, which may be attributed to altered intracellular polysaccharide construction.


How to Tackle Key Challenges in the Promotion of Physical Activity among Older Adults (65+): The AEQUIPA Network Approach.
The paper introduces the theoretical framework and methods/instruments used by the Physical Activity and Health Equity: Primary Prevention for Healthy Ageing (AEQUIPA) prevention research network as an interdisciplinary approach to tackle key challenges in the promotion of physical activity among older people (65+). Drawing on the social-ecological model, the AEQUIPA network developed an interdisciplinary methodological design including quantitative/qualitative studies and systematic reviews, while combining expertise from diverse fields: public health, psychology, urban planning, sports sciences, health technology and geriatrics. AEQUIPA tackles key challenges when promoting physical activity (PA) in older adults: tailoring of interventions, fostering community readiness and participation, strengthening intersectoral collaboration, using new technological devices and evaluating intervention generated inequalities. AEQUIPA aims to strengthen the evidence base for age-specific preventive PA interventions and to yield new insights into the explanatory power of individual and contextual factors. Currently, the empirical work is still underway. First experiences indicate that thenetwork has achieved a strong regional linkage with communities, local stakeholders and individuals. However, involving inactive persons and individuals from minority groups remained challenging. A review of existing PA intervention studies among the elderly revealed the potential to assess equity effects. The results will add to the theoretical and methodological discussion on evidence-based age-specific PA interventions and will contribute to the discussion about European and national health targets.


Design and self-assembly of ditopic and tetratopic cavitand complexes.
The self-assembly of open ditopic and tetratopic cavitand complexes has been investigated by using monofunctionalized cavitand ligands and suitable metal precursors. In the case of ditopic complexes, self-assembly protocols, leading exclusively to the formation of both thermodynamically stable cis-Pt square-planar complexes 8 and 9 and the kinetically inert fac-Re octahedral complex 14, have been elaborated. The use of cis-[Pt(CH3)CN)2Cl2] as metal precursor led to the formation of monotopic trans-10 and ditopic trans-11 cavitand complexes, while cis-[Pt(dmso)2Cl2] afforded both cis-13 and trans-11 isomers. The self-assembly of tetratopic cavitand complexes has been achieved by using mononuclear [Pd(CH3CN)4(BF4)2] and dinuclear [M2(tppb)(OTf)4] (19: M = Pt; 20: M = Pd) metal precursors. Only the tetratopic dinuclear complexes 21 and 22 were stable. The ligand configuration with two phosphorus and two cavitand ligands at the metal centers is the most appropriate to build tetratopic cavitand complexes with sufficient kinetic stability.


A clinical scoring system to predict the development of bronchopulmonary dysplasia.
This study aims to develop a scoring system for the prediction of bronchopulmonary dysplasia (BPD). Medical records of 652 infants whose gestational age and birth weight were below 32 weeks and 1,500 g, respectively, and who survived beyond 28th postnatal day were reviewed retrospectively. Logistic regression methods were used to determine the clinical and demographic risk factors within the first 72 hours of life associated with BPD, as well as the weights of these factors on developing BPD. Predictive accuracy of the scoring system was tested prospectively at the same unit. Birth weight, gestational age, gender, presence of respiratory distress syndrome, patent ductus arteriosus, intraventricular hemorrhage, hypotension were the most important risk factors for BPD. Therefore, a scoring system (BPD-TM score) ranging from 0 to 13 and grouped in four tiers (0-3: low, 4-6: low intermediate, 7-9: high intermediate, and 10-13: high risk) was developed based on these factors. Below the score of 4, 4.1% of infants (18/436), above the score of 9, 100% (29/29) of the infants developed BPD. The score was validated successfully in 172 infants. With this easy to use scoring system, one can predict the neonate at risk for BPD at 72 hours of life and direct preventive measures toward these infants.


Hypoxic excitation in neurons cultured from the rostral ventrolateral medulla of the neonatal rat.
Neurons within cardiorespiratory regions of the rostral ventrolateral medulla (RVLM) have been shown to be excited by local hypoxia. To determine the electrophysiological properties of these excitatory responses to hypoxia, we developed a primary dissociated cell culture system to examine the intrinsic response of RVLM neurons to hypoxia. Neonatal rat neurons plated on medullary astrocyte monolayers were studied using the whole cell perforated patch-clamp technique. Sodium cyanide (NaCN, 0.5-10 mM) was used, and membrane potential (V(m)), firing frequency, and input resistance were examined. In 11 of 19 neurons, NaCN produced a V(m) depolarization, an increase in firing frequency, and a decrease in input resistance, suggesting the opening of a cation channel. The hypoxic depolarization had a linear dose response and was dependent on baseline V(m), with a greater response at more hyperpolarized V(m). In 8 of 19 neurons, NaCN produced a V(m) hyperpolarization, decrease in firing frequency, and variable changes in input resistance. The V(m) hyperpolarization exhibited an all-or-none dose response and was independent of baseline V(m). These differential responses to NaCN were retained after synaptic blockade with low Ca(2+)-high Mg(2+) or TTX. Thus hypoxic excitation 1) is maintained in cell culture, 2) is an intrinsic response, and 3) is likely due to the increase in a cation current. These hypoxia-excited neurons are likely candidates to function as central oxygen sensors.


Rapid-freezing transmural cardiac biopsy drill.
We have built a transmural cardiac biopsy drill that uses an air turbine (10,000 rpm) to turn a stainless steel bore of either 2.0 or 4.5 mm diam. Vacuum draws the cut biopsies through the drill into isopentane chilled to -150 degrees C with liquid nitrogen. The steel bores cut through the beating canine left ventricular free wall in 0.14 +/- 0.04 (SD) permitting sampling in discrete portions of the cardiac cycle. Small and large biopsies traverse the drill in 0.48 +/- 0.19 and 0.15 +/- 0.04 s, respectively. Large biopsies freeze in 1.46 +/- 0.73 s, whereas small biopsies are calculated to freeze in 0.28 s. Average biopsy weights are 34 +/- 14 and 180 +/- 71 mg. Left ventricular myocardial blood flow measured with radionuclide-labeled microspheres showed that muscle plugs sutured into wound sites of 4.5-mm-diam biopsies cause a 30% reduction in myocardial blood flow within 0.5 cm of the biopsy site. Light microscopy shows normal cardiac muscle with little damage from drill rotation.


Parenting from a County Jail: Parenting from Beyond the Bars.
The incarceration of mothers affects 1.7 million minor children who are at high risk of behavioral problems, emotional trauma, and future incarceration. The jail setting removes women from society for brief periods of time providing an opportunity to offer essential interventions for those who will be returning to the community, and possibly their children, rather quickly. Utilizing a pre- and post-test quasi-experimental design, we measured the self-reported treatment effects for female inmates, housed in a county jail in the New York metropolitan area, who completed an evidence-based parenting program. Program participants reported reduced depression, increased parental knowledge, and increased communication with their families and fellow inmates. The results indicated that alternative ways of measuring program success are needed - particularly for populations who present with long histories of criminal justice involvement and drug use.


The efficacy of visual therapy: accommodative disorders and non-strabismic anomalies of binocular vision.
Questions concerning the effectiveness of therapeutic intervention designed to modify the function of an individual's visual system have been raised by professional groups, consumer groups, insurance carriers, students, educators, as well as "pure" scientists. This paper examines the available literature in order to answer the question, "Is there evidence that 'orthoptics' or 'vision therapy' causes changes in an individual's accommodative or vergence eye movement systems?" This review neither examines alternative methods of causing these changes nor provides information concerning which particular techniques are most effective, although the literature does provide such information. Certain conditions are explained to aid the student and lay-reader in understanding the literature cited. The literature cited substantiates that visual therapy can modify visual functions and also points out the relationship of these changes to the relief of certain symptoms.


Mechanism of inhibition of bacteriophage T7 RNA polymerase by T7 lysozyme.
Bacteriophage T7 lysozyme is known to inhibit transcription by T7 RNA polymerase. Lysozyme present before initiation inhibited the synthesis of long RNA chains but did not inhibit elongation when added shortly after chains were initiated. A combination of gel-shift and transcription assays showed that lysozyme and polymerase form a 1:1 complex that binds promoter DNA and makes abortive transcripts, indicating that lysozyme has little effect on the early steps of transcription. Extension of stalled transcription complexes suggested that a transcribing polymerase becomes resistant to lysozyme inhibition after synthesis of an RNA chain as short as 15 nucleotides. It seems likely that bound lysozyme prevents an initiating polymerase from converting to an elongation complex. This conversion is thought to involve both a conformational change in the polymerase and the binding of nascent RNA. Gel-shift experiments indicated that lysozyme does not interfere with the binding of RNA, so it probably prevents a necessary conformational change in the polymerase. Lysozyme also increased pausing or termination at two sites in lambda DNA and at a site near the right end of the concatemer junction of T7 DNA. If pausing at these sites involves a reversal from the elongation to the initiation conformation, lysozyme may increase pausing or termination by "locking in" the initiation conformation. The arrest of transcription complexes near promoters and near the right end of the concatemer junction almost certainly must relate to lysozyme's ability to stimulate replication, maturation and packaging of T7 DNA during T7 infection.


Alaskan Berry Extracts Promote Dermal Wound Repair Through Modulation of Bioenergetics and Integrin Signaling.
Various wild berry species endemic to Alaska and the circumpolar North that exhibit unique medicinal properties have long been appreciated by indigenous Arctic communities. Traditional use of Alaskan berry preparations in the treatment of skin wounds is recorded but has not been scientifically evaluated. Alaskan wild berries feature diverse phytochemical compositions that contain a variety of bioactive polyphenols exhibiting anti-inflammatory, antioxidant, and antimicrobial properties, making them ideal for wound healing interventions and natural anti-aging cosmeceutical formulations. Given increasing interest in identifying biologically active plant constituents for wound care and cosmeceutical applications, the objective of this study was to screen several wild berry species endemic to Alaska and the circumpolar Artic for wound healing and in the crude, polyphenol-enriched, and further fractionated extracts of: Empetrum nigrum (crowberry), Vaccinium uliginosum (bog blueberry), and V. vitis-idaea (low-bush cranberry or lingonberry). A cell migration assay with human dermal fibroblasts (HDFa) was performed to model promotion of wound closure, revealing that bog blueberry extract most actively promoted migration, whereas divergent effects observed with other berry extracts were related to compositional disparities. Lipopolysaccharide (LPS)-stimulated inflammatory response variables measured in RAW 264.7 macrophages [reactive oxygen species (ROS), NO production, prostaglandin-endoperoxide synthase 2 (COX-2), and inducible nitric oxide synthase (iNOS) expression] were suppressed by most extracts/fractions, but especially bog blueberry and proanthocyanidin (PAC) fractions. Wild berry germplasm contained abundant complex flavonoid structures such as PAC and anthocyanins (ANCs), associated with enhanced repair and inflammatory resolution in these models. Next, underlying mechanisms by which PACs and bioactive metabolites (B2 dimer and epicatechin) could influence wound repair and tissue regeneration were examined. PAC metabolites promoted scratch-wound closure and appeared to exert the highest impacts on early stages of wound healing through stimulating mitochondrial bioenergetics (basal respiration, ATP production, and maximum respiratory capacity) and upregulating expression of important extracellular matrix (ECM) proteins (integrin-ß1 and collagen type I α2 chain). Targeting cellular bioenergetics and integrin-mediated cell-ECM signaling with bioactives from Alaskan wild berries shows considerable therapeutic promise to treat chronic skin wounds and inflammatory skin disorders, as well as more generally to support regenerative healing responses and restore function in a variety of tissue and organ settings after injury or aging.


Searching the Porphyromonas gingivalis genome with peptide fragmentation mass spectra.
An approach is described for genomic database searching based on experimentally observed proteolytic fragments, e.g., isolated from 1D or 2D gels or analyzed directly, that can be applied to unfinished prokaryotic genomic data in the absence of annotations or previously assigned open reading frames (ORFs). This variation on the database search is in contrast to the more familiar use of peptide mass spectral fragmentation data to search fully annotated inferred protein databases, e.g., OWL or SWISS-PROT. We compared the SEQUEST search results from a six reading frame translation of the Porphyromonas gingivalis genome DNA sequence with those from computationally derived ORFs created using publicly available genomics software tools. The ORF approach eliminated many of the artifacts present in output from the six reading frame search. The method was applied to uninterpreted tandem mass spectrometric data derived from proteins secreted by the periodontal pathogen Porphyromonas gingivalis in response to the gingival epithelial cell environment, a model system for the study of host-pathogen interactions relevant to human periodontal disease.


Recursive implementation of a two-step nonparametric decision rule.
The two-step approach to nonparametric discrimination is that of estimating class-conditional densities and deriving the Bayes decision rule as if the estimates were true. Direct implementation of such a decision rule ecounters two computational problems. Complexity increases with sample size, and finite precision limits the decision rule domain. Here a recursive algorithm to reduce the expected number of operations and word-length limitations below that of the direct approach is developed. A special case of the formulation reduces to the weighted k-nearest-neighbor rule.


Peroxidase as a developmental marker in plant tissue culture.
Peroxidase was studied as a developmental marker in pumpkin (Cucurbita pepo L.) callus lines and horse-radish (Armoracia lapathifolia Gilib) transformants. Embryogenic callus lines DE grown on MS medium with 2.4-D and NA-3 grown on medium with NAA and adenine sulfate showed about a 20 times higher enzyme activity than the habituated non-embryogenic line Z5b/T grown on medium without hormones. A rise in peroxidase activity indicated that somatic embryogenesis was triggered in a few habituated tissue cultures. Separated globular embryoids had a manifold lower enzyme activity than the callus from which they originated. SDS-electrophoresis showed distinct polypeptide patterns between the horse-radish leaves and crown galls, but the tumor characteristic protein bands failed to be identified. In horse-radish crown galls and short bushy plants regenerated from hairy roots an enhanced peroxidase activity was registered. Due to its high peroxidase level and abundant biomass production horse-radish transformants should facilitate enzyme production.


Comparison of the structure and aspects of the proteinase-binding properties of cystic fibrotic alpha-macroglobulin with normal alpha 2-macroglobulin.
Considerable attention has been focused recently on alpha 2-macroglobulin (alpha 2M), a major endopeptidase inhibitor in blood plasma, as a possible source of the primary defect in cystic fibrosis (CF). We report here studies designed to compare the structure of CF alpha 2M with normal alpha 2M to determine if there is a difference. The physicochemical properties of purified alpha 2M as revealed by various electrophoretic techniques, covalent proteinase binding properties, and primary structural studies on a variety of partial hydrolyzates of CF alpha 2M and normal alpha 2M are compared. These studies were carried out on eight different individual isolates of CF alpha 2M and three age-matched normal alpha 2M preparations and alpha 2M isolated from fetal cord blood. Three properties of CF alpha 2M were studied by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE): (1) the existence of four identically-sized subunits in the native molecule (10), (2) the cleavage of this subunit into fragments of approximately 100,000 daltons upon interaction with proteinases (10), and (3) the cleavage of an alkaline/heat sensitive bond to produce 120,000 and 60,000 dalton fragments (11). Both CF and normal alpha 2M were cleaved to the extent of 79-87%, CF alpha 2M behaves identically with normal alpha 2M with regard to all these properties. Salvesen and Barrett (24) have demonstrated that varying proportions of several [125I]-labeled proteinases form SDS-stable, non-reducible links to normal alpha 2M. Two of the CF alpha 2M preparations were studied to determine if similar covalent binding of proteinases occurred. The positions of the labeled and % of proteinase bound bands in SDS/reduced PAGE system were identical for normal alpha 2M and CF alpha 2M. These results indicate that CF alpha 2M behaves normally with regard to covalent binding of proteinases. Qualitative comparison of the peptide fragments separated by SDS-PAGE or isoelectric focusing of CF and normal alpha 2M produced by partial proteolysis with trypsin, chymotrypsin or Staphylococcus aureus V-8 proteinase did not reveal any difference unique to CF alpha 2M. The cyanogen bromide fragmentation studies and the cysteine cleavage studies also indicated that no major change in the positions of methionyl residues or cysteinyl/cystinyl residues has occurred in CF alpha 2M. The failure of all these different studies and those reported by others to demonstrate any differences between CF and normal alpha 2M makes it highly unlikely that there is a primary defect in alpha 2M in CF.


Syndecan 4 interacts genetically with Vangl2 to regulate neural tube closure and planar cell polarity.
Syndecan 4 (Sdc4) is a cell-surface heparan sulfate proteoglycan (HSPG) that regulates gastrulation, neural tube closure and directed neural crest migration in Xenopus development. To determine whether Sdc4 participates in Wnt/PCP signaling during mouse development, we evaluated a possible interaction between a null mutation of Sdc4 and the loop-tail allele of Vangl2. Sdc4 is expressed in multiple tissues, but particularly in the non-neural ectoderm, hindgut and otic vesicles. Sdc4;Vangl2(Lp) compound mutant mice have defective spinal neural tube closure, disrupted orientation of the stereocilia bundles in the cochlea and delayed wound healing, demonstrating a strong genetic interaction. In Xenopus, co-injection of suboptimal amounts of Sdc4 and Vangl2 morpholinos resulted in a significantly greater proportion of embryos with defective neural tube closure than each individual morpholino alone. To probe the mechanism of this interaction, we overexpressed or knocked down Vangl2 function in HEK293 cells. The Sdc4 and Vangl2 proteins colocalize, and Vangl2, particularly the Vangl2(Lp) mutant form, diminishes Sdc4 protein levels. Conversely, Vangl2 knockdown enhances Sdc4 protein levels. Overall HSPG steady-state levels were regulated by Vangl2, suggesting a molecular mechanism for the genetic interaction in which Vangl2(Lp/+) enhances the Sdc4-null phenotype. This could be mediated via heparan sulfate residues, as Vangl2(Lp/+) embryos fail to initiate neural tube closure and develop craniorachischisis (usually seen only in Vangl2(Lp/Lp)) when cultured in the presence of chlorate, a sulfation inhibitor. These results demonstrate that Sdc4 can participate in the Wnt/PCP pathway, unveiling its importance during neural tube closure in mammalian embryos.


[The unicorn and the pharmacists. Early modern views on the presumed anti-toxic effects of unicorn horn].
Around 1600, the age-old belief in the anti-toxic effect of unicorn horn began to be called into question. This is evidenced by the views of two well-known French pharmaceutic authorities whose publications are discussed in this paper: the surgeon Ambroise Paré (1510-1590), court physician to four French kings, and the Montpellier pharmacist Laurent Catelan (1568-1647), who owned a famous cabinet de curiosités. Although Paré had to accept, however reluctantly, the existence of the unicorn (since it is mentioned in the Bible), he vehemently denied the supposed medicinal effect of unicorn products. He defended his position by an appeal to ancient and contemporary authorities, by rational argumentation, and by experiment. Paré's arguments failed to convince Catelan, who adhered to an alternative, so-called spagyric, medical theory of neoplatonic inspiration, as propagated by Paracelsus and Ficino. Catelan remained convinced of the efficacity of unicorn horn, which in his view could drain the human body from any poisonous substance. The medical establishment being reluctant to give up a rewarding source of income,'unicorn' remained much in demand as a prescription.


CXCR4 antagonist AMD3100 attenuates colonic damage in mice with experimental colitis.
To investigate the effects of the chemokine stromal cell-derived factor-1 (CXCL12) receptor (CXCR4) antagonist AMD3100 on colonic inflammation and epithelial barrier in dextran sulfate sodium (DSS)-induced colitis in mice. Experimental colitis was induced by administration of 5% DSS for 7 d, and assays performed on intestinal segments from the ileocecal valve to the anus. Colonic morphology was examined by hematoxylin and eosin staining. Colonic cytokines were determined by enzyme-linked immunosorbent assay. Myeloperoxidase (MPO) activity (indicator of inflammatory infiltration) was observed spectrophotometrically. Gut permeability was assessed by mucosal-to-serosal clearance of fluorescein isothiocyanate-conjugated dextran 4000 (FD4) in everted gut sacs. The apoptosis of colonic epithelium was assessed by Hoechst-33342 staining. To further elucidate the role of CXCR4 in colonic inflammation, we also investigated the effect of AMD3100 on migration and cytokine production of isolated peripheral blood mononuclear cells (PBMCs). DSS-induced colitis was characterized by morphologic changes, as well as increased colonic cytokines, inflammatory infiltration, epithelial apoptosis, and intestinal permeability in mice. In AMD3100-treated mice, epithelial destruction, inflammatory infiltration, and submucosal edema were markedly reduced; colonic tumor necrosis factor-alpha (TNF-alpha), interleukin-6 (IL-6) and interferon-gamma (IFN-gamma) levels, as well as MPO activity were significantly decreased. Increased intestinal permeability in DSS-treated mice was significantly reduced by AMD3100. The number of apoptotic cells in colitis mice was markedly increased after DSS administration, and decreased when treated with the CXCR4 antagonist AMD3100. In pre-activated PBMCs, CXCL12 stimulation significantly increased the migration of PBMCs, and was inhibited by AMD3100. Moderately increased TNF-alpha, IL-6, and IFN-gamma from CXCL12-treated PBMCs were also reduced by AMD3100. The CXCR4 antagonist AMD3100 exerts therapeutic effects on experimental colitis by inhibiting colonic inflammation and enhancing epithelial barrier integrity.


Pain during PICC insertion in preterm infants: the needle is not the only problem.
To evaluate whether warm povidone-iodine (PI) application before peripherally inserted central catheter (PICC) placement eased pain related to the procedure in premature infants and reduced the duration of the procedure and the number of attempts. A prospective randomized controlled trial was conducted with infants born before 32 weeks of gestation who required the first placement of the PICC. Skin disinfection was performed with warm PI before the procedure in the warm PI(W-PI) group, whereas PI kept at room temperature was used in the regular PI(R-PI) group. NPASS scores of the infants were evaluated three times: at baseline(T0), during skin preparation(T1), and during needle insertion(T2). Fifty-two infants (26 in the W-PI group,26 in the R-PI group) were enrolled in the study. The perinatal and baseline demographic characteristics did not significantly differ between the two groups. While the median NPASS scores at T0 and T2 were similar between the groups, the median T1 score was significantly higher in the R-PI group(p = .019). While the median NPASS scores at T1 and T2 were similar in the R-PI group, there was a significant difference in the W-PI group, with NPASS scores being significantly lower at T1 compared to T2. The results demonstrate that skin disinfection was just as painful as needle insertion in the R-PI group. The duration of the procedure and the number of needle insertions were significantly lower in the W-PI group. Before invasive interventions, such as PICC insertion, we recommend warm PI as a part of non-pharmacological pain management.