Phenotypic analysis of the ccp1Delta and ccp1Delta-ccp1W191F mutant strains of Saccharomyces cerevisiae indicates that cytochrome c peroxidase functions in oxidative-stress signaling.
Yeast cytochrome c peroxidase (CCP) efficiently catalyzes the reduction of H(2)O(2) to H(2)O by ferrocytochrome c in vitro. The physiological function of CCP, a heme peroxidase that is targeted to the mitochondrial intermembrane space of Saccharomyces cerevisiae, is not known. CCP1-null-mutant cells in the W303-1B genetic background (ccp1Delta) grew as well as wild-type cells with glucose, ethanol, glycerol or lactate as carbon sources but with a shorter initial doubling time. Monitoring growth over 10 days demonstrated that CCP1 does not enhance mitochondrial function in unstressed cells. No role for CCP1 was apparent in cells exposed to heat stress under aerobic or anaerobic conditions. However, the detoxification function of CCP protected respiring mitochondria when cells were challenged with H(2)O(2). Transformation of ccp1Delta with ccp1(W191F), which encodes the CCP(W191F) mutant enzyme lacking CCP activity, significantly increased the sensitivity to H(2)O(2) of exponential-phase fermenting cells. In contrast, stationary-phase (7-day) ccp1Delta-ccp1(W191F) exhibited wild-type tolerance to H(2)O(2), which exceeded that of ccp1Delta. Challenge with H(2)O(2) caused increased CCP, superoxide dismutase and catalase antioxidant enzyme activities (but not glutathione reductase activity) in exponentially growing cells and decreased antioxidant activities in stationary-phase cells. Although unstressed stationary-phase ccp1Delta exhibited the highest catalase and glutathione reductase activities, a greater loss of these antioxidant activities was observed on H(2)O(2) exposure in ccp1Delta than in ccp1Delta-ccp1(W191F) and wild-type cells. The phenotypic differences reported here between the ccp1Delta and ccp1Delta-ccp1(W191F) strains lacking CCP activity provide strong evidence that CCP has separate antioxidant and signaling functions in yeast.


[Clinical course of brain tumors in childhood].
Brain tumors are, after leukemias, the most frequent fatal neoplasms of infancy. The clinical features and symptoms are often markedly different from those observed in the adult forms, according to the peculiar anatomy and behaviour of the child. Persistent headache, vomiting, astenia , behavioral alterations may be the precocious findings. Later, some more specific and suggestive signs such as strabismus, dyplopia , fast head size increase, funduscopic alterations, ataxia, paresis and nystagmus may be observed. On their appearance a prompt diagnostic work-up should be performed. The tumors of the posterior fossa (cerebellar astrocytoma and medulloblastoma, brainstem glyoma , hependimoma in decreasing order of frequency) generally cause precocious symptoms because of the small dimension of the subtentorial space; the presence in this region of several fundamental nuclei and pathways may explain how also small tumors may cause severe deficits. Supratentorial tumors (astrocytoma, malignant glyoma , hependimoma , craniofaringioma ) often show a more prolonged latency and may begin with signs of endocranial hypertension, seizures, or sometimes with hormonal troubles according to the involved anatomic structures. Hypothalamic astrocytoma is responsible of an extreme weight loss as far as to a cachetic status, due to the hyperincretion of GH. Finally, plexus papilloma, dermoid, optic nerve glyoma , oligodendroglyoma , germinoma, teratoma are responsible of a small number of child brain tumors, with different localization and symptomatology.


A novel tool for B-cell tolerance research: characterization of mouse alloantibody development using a simple and reliable cellular ELISA technique.
In animal-based transplantation research, the measurement of anti-donor antibodies in transplant recipients is limited by lack of an appropriate technique. We have developed a novel immunoassay capable of quantifying antibody bound to cell-surface major histo- compatability complex (MHC) and non-MHC antigens, using splenocytes from wild-type and MHC-deficient mice as antigen-bearing target cells. We utilized our "cellular ELISA" (CELISA) technique to study the development of tolerance versus immunity in the B-cell compartment in response to neonatal exposure to allogeneic fetal liver cells (FLC). This neonatal tolerance protocol typically induces permanent acceptance of donor-type and third-party cardiac allografts, but rejection of both donor-type and third-party skin grafts occurs. C3H/He (C3H; H-2(k)) mice were injected as neonates with BALB/c (BALB; H-2(d)) FLC and transplanted as adults with C57BL/6 (B6; H-2(b)) cardiac grafts. Despite long-term acceptance of third-party B6 cardiac grafts, serum contained increased anti-B6 IgG and IgM levels as measured by CELISA; IgM production was elevated by 2 weeks posttransplant and remained stable, while IgG production increased rapidly between 2 and 5 weeks posttransplant. In another experimental setting, CELISA assays were able to detect that neonatal injection of C3H mice with FLC from wild-type B6 mice or from MHC class II-deficient or class I/II-deficient (B6 background) mice (CI(+)CII(+), CI(+)CII(-), CI(-)CII(-), respectively) prevented sensitization to B6 antigens by subsequent skin transplants but did not induce graft acceptance, whereas FLC from MHC class I-deficient-only (CI(-)CII(+)) did not prevent B6 sensitization. The CELISA technique is a simple and sensitive means for quantifying alloantibodies in mice and will assist in further delineating the role of the B-cell compartment in neonatally induced cardiac allograft acceptance.


8-Hydroxyquinoline benzoates as highly sensitive fluorescent chemosensors for transition metal ions.
[graph: see text] 8-Hydroxyquinoline benzoates were developed as a new set of 8-HQ derivatives for highly sensitive fluorescent chemosensors for transition metal ions. A prominent fluorescence enhancement was found in the presence of transition metal ions such as Hg2+ and Cu2+, and this was suggested to result from the suppression of radiationless transitions from the npi state in the chemosensors.


Revision of the soft scale genus Paralecanium (Hemiptera: Coccomorpha: Coccidae) with the introduction of three new genera and twenty new species.
The soft scale insect genus Paralecanium Cockerell (Hemiptera: Coccomorpha: Coccidae), mainly characterised by the possession of fan-shaped marginal setae, has never been revised and most of the species currently recognised were described in the early part of the twentieth century. The present revision is based on the morphology of the adult females and redescribes and illustrates all 34 species or subspecies previously included in the genus. These three subspecies have been raised to full specific rank, namely P. expansum javanicum (Green) to P. javanicum (Green) stat. n.; P. expansum rotundum (Green) to P. rotundum (Green) stat. n. and P. frenchii macrozamiae (Fuller) to P. macrozamiae (Fuller) stat. n. Of the remaining previously recognised species, three have been made junior synonyms: P. album Takahashi is here considered to be a junior synonym of P. metallicum (Green), syn. n.; P. angkorense Takahashi is considered to be a junior synonym ofP. cocophyllae Banks, syn. n.; and P. limbatum Green is considered to be a junior synonym of P. geometricum (Green), syn. n. Paralecanium marianum Cockerell from Brazil is clearly not closely related to Paralecanium and so a new genus, Mariacoccus Hodgson Williams gen. n., has been introduced to take it and the adult female is redescribed and illustrated; comparison of the adult female of M. marianus with that of Coccus lizeri (Fonseca) showed that the latter species is a junior synonym of M. marianus (Cockerell), syn. n. Based on the morphology of the above species and on the new species mentioned below, two new genera have been introduced: Insularicoccus Hodgson Williams gen. n. (type species P. carolinensis Beardsley), and Discochiton Hodgson Williams gen. n. (type species D. martini Hodgson, spec. n.). This brings the number of soft scale genera with fan-shaped marginal setae to three. Diagnoses of each of these 3 genera are presented, along with a key to the genera based on adult female morphology. The species that have been left in Paralecanium Cockerell are: P. calophylli (Green), P. frenchii (Maskell), P. geometricum (Green), P. hainanense Takahashi, P. machili Takahashi, P. macrozamiae (Fuller), P. maculatum Takahashi, P. marginatum (Green), P. maritimum (Green), P. minutum Takahashi, P. neomaritimum Takahashi, P. ovatum Morrison, P. pahanense Takahashi, P. peradeniyense (Green), P. planum (Green) and P. zonatum (Green). The species transferred to Discochiton as comb. n. are: P. album Takahashi, P. cocophyllae Banks, P. expansum (Green), P. javanicum (Green), P. luzonicum Cockerell, P. malainum Takahashi, P. mancum (Green), P. metallicum (Green), P. milleri Takahashi, P. pseudexpansum (Green), P. quadratum (Green), P. rotundum (Green),P. trifasciatum (Green) and P. vacuum Morrison. The only species transferred to Insularicoccus is the type species, namelyP. carolinense (Beardsley), comb. n. In addition, adult females of the following 11 new species are described and illustrated in the genus Paralecanium (as defined here) spec. n.: P. acinaces Hodgson, P. busoense Hodgson, P. claviseta Hodgson, P. comperei Hodgson, P. cypripedium Hodgson, P. elongatum Hodgson, P. leei Hodgson, P. morobeense Hodgson, P. neoguineense Hodgson, P. palawanense Hodgson and P. vacerra Hodgson. In addition, the following 8 species are described and illustrated as Discochiton spec. n.: D. browni Hodgson, D. crenulatum Hodgson, D. diplodiscus Hodgson, D. martini Hodgson, D. papillatum Hodgson, D. paucipedis Hodgson, D. sarawakense Hodgson and D. seychellarum Williams Hodgson, and 1 species of Insularicoccus is described as new and illustrated, namely I. syzygium Hodgson spec. n. Keys, based on adult female morphology, are provided for separation of all the species in each genus. As part of this revision, the morphological characters used to diagnose species in this group of genera are re-evaluated and a number of new characters found; greater emphasis has been placed on some character-states, so the basic morphology of this group of genera is also described. Lectotypes have been designated for 22 species, namely: Lecanium calophylli Green; L. expansum Green; L. expansum javanicum Green; L. expansum metallicum Green; L. expansum rotundum Green; rotundum Green; L. geometricum Green; L. limbatum Green; L. mancum Green; L. marginatum Green; L. maritimum Green; L. peradeniyense Green; L. planum Green; L. pseudexpansum Green; L. quadratum Green; L. trifasciatum Green and L. zonatum Green. Also: Paralecanium album Takahashi; P. hainanense Takahashi; P. malianum Takahashi; P. marianum Cockerell; P. neomaritimum Takahashi and P. vacuum Morrison. Finally, an example of the first-instar nymphs, second-instar males and second/third instar females of both Paralecanium and Discochiton are described and illustrated and compared with those already known in the tribe Paralecaniini.


Use of spherical harmonic deconvolution methods to compensate for nonlinear gradient effects on MRI images.
Spatial encoding in MR techniques is achieved by sampling the signal as a function of time in the presence of a magnetic field gradient. The gradients are assumed to generate a linear magnetic field gradient, and typical image reconstruction relies upon this approximation. However, high-speed gradients in the current generation of MRI scanners often sacrifice linearity for improvements in speed. Such nonlinearity results in distorted images. The problem is presented in terms of first principles, and a correction method based on a gradient field spherical harmonic expansion is proposed. In our case, the amount of distortion measured within a typical field of view (FOV) required for head imaging is sufficiently large that without the use of some distortion correction technique, the images would be of limited use for stereotaxy or longitudinal studies, where precise volumetric information is required.


Predicting the result of additional second-trimester markers from a woman's first-trimester marker profile: a new concept in Down syndrome screening.
To describe a method for deciding whether an individual's first-trimester Down syndrome screening test result justifies further testing in the second trimester. Statistical modelling was used to estimate the distribution of second-trimester marker profiles for a given first-trimester profile and hence the probability of a final positive result, using a 1 in 250 term cut-off. A multi-variate log Gaussian model was used with published parameters. Markers were maternal serum pregnancy-associated plasma protein-A and free beta-human chorionic gonadotrophin (hCG) at 10 weeks, nuchal translucency at 11 weeks, and second-trimester maternal serum alpha-fetoprotein, total hCG, unconjugated estriol and inhibin-A. To illustrate the method, the model was applied to a published series of 24 Down syndrome and 367 unaffected pregnancies. Modelling predicts that for 63% Down syndrome and 0.4% unaffected pregnancies having first-trimester tests, there is a 50% or more probability of a final positive result. A step-wise sequential screening policy based on immediate prenatal diagnosis for those with high probability and second-trimester testing for the remainder would have a 90% detection rate and 1.7% false-positive rate. Modelling also predicts 8.0% Down syndrome and 89% unaffected pregnancies with probabilities below 3%. A contingent screening policy restricting second-trimester testing to those with 3-49% probabilities would have an 88% detection rate and 1.4% false-positive rate. Predicting the probability of a positive final result from the first-trimester marker profile has potential utility, either as a decision aide for individual women or as a formal part of screening policy in selecting a subset of women for second-trimester testing.


Translesion DNA Synthesis.
All living organisms are continually exposed to agents that damage their DNA, which threatens the integrity of their genome. As a consequence, cells are equipped with a plethora of DNA repair enzymes to remove the damaged DNA. Unfortunately, situations nevertheless arise where lesions persist, and these lesions block the progression of the cell's replicase. In these situations, cells are forced to choose between recombination-mediated "damage avoidance" pathways or a specialized DNA polymerase (pol) to traverse the blocking lesion. The latter process is referred to as Translesion DNA Synthesis (TLS). As inferred by its name, TLS not only results in bases being (mis)incorporated opposite DNA lesions but also bases being (mis)incorporated downstream of the replicase-blocking lesion, so as to ensure continued genome duplication and cell survival. Escherichia coli and Salmonella typhimurium possess five DNA polymerases, and while all have been shown to facilitate TLS under certain experimental conditions, it is clear that the LexA-regulated and damage-inducible pols II, IV, and V perform the vast majority of TLS under physiological conditions. Pol V can traverse a wide range of DNA lesions and performs the bulk of mutagenic TLS, whereas pol II and pol IV appear to be more specialized TLS polymerases.


[Water intake of primary and middle school students in four cities of China].
To investigate total intake of drinking water of primary and middle school students in four cities of China, and to provide scientific evidence for developing standard of intake of drinking water for primary and middle school students in China. A total of 5914 primary and middle school students were selected from Beijing, Shanghai, Chengdu and Guangzhou using multi-stage random sampling method and 5868 subjects completed the survey. The information on the amounts of daily drinking water was recorded for seven consecutive days using a 24 hour measurement. The amounts of daily drinking water among different cities and ages and between boys and girls or urban and rural were analyzed. The average daily total drinking water of subjects was (1089 ± 540) ml, with significant differences among the four cities ((1165 ± 549), (1126 ± 544), (866 ± 421), (1185 ± 568) ml in Beijing, Shanghai, Chengdu and Guangzhou, respectively; F = 114.28, P < 0.05). The average daily drinking water was significantly higher in boys (1157 ± 575) ml than in girls (1026 ± 498) ml (Z = 8.88, P < 0.05), and in urban (1185 ± 578) ml than in rural (991 ± 480) ml (Z = 13.94, P < 0.05). The difference among different ages was statistically significant (F = 91.53, P < 0.05) with ascending order in 7 - 10, 11 - 13 and 14 - 17 years old ((953 ± 483), (1134 ± 551) and (1170 ± 557) ml respectively). The daily consumption of total drinking water is different in primary and middle school students among different cities, regions, genders and ages.


Diagnostic value of lumbar puncture among infants and children presenting with fever and convulsions.
Central nervous system (CNS) infections can be categorized according to the nature of the infectious pathogen into viral, bacterial, protozoan, or fungal. The diagnosis of diffuse CNS infections depends on examination of cerebrospinal fluid (CSF) obtained by lumbar puncture (LP). The aim of this work was to determine the diagnostic value of CSF analysis in infants and children presenting with fever and convulsions. Detailed clinical data of infants and children included in this study were collected with special reference to the course and duration of the illness, description of the convulsions, consciousness level, signs of increased intracranial pressure, and signs of meningeal irritation. Lumbar puncture and chemical and bacteriological analyses of the obtained cerebrospinal fluid were done for all of the children. The total number of children included in the study was 85, they had a median age 19 months, and 88% of them had generalized convulsions. CSF examination revealed that 20% had abnormal physical findings, while 23.5% had abnormal white blood cell count (WBC) (CSF Pleocytosis). CSF cultures were done in three cases with the highest White blood cells (WBCs), and streptococcus pneumoniae was present in all three cases. This study found that CNS infections are not uncommon in infants and children presenting with fever and convulsions in our locality, and acute bacterial meningitis cannot be excluded. However, its presence in the absence of clinical symptoms and signs of meningeal irritation is a remote possibility, but it should always be considered.


The colonial context of Filipino American immigrants' psychological experiences.
Because of the long colonial history of Filipinos and the highly Americanized climate of postcolonial Philippines, many scholars from various disciplines have speculated that colonialism and its legacies may play major roles in Filipino emigration to the United States. However, there are no known empirical studies in psychology that specifically investigate whether colonialism and its effects have influenced the psychological experiences of Filipino American immigrants prior to their arrival in the United States. Further, there is no existing empirical study that specifically investigates the extent to which colonialism and its legacies continue to influence Filipino American immigrants' mental health. Thus, using interviews (N = 6) and surveys (N = 219) with Filipino American immigrants, two studies found that colonialism and its consequences are important factors to consider when conceptualizing the psychological experiences of Filipino American immigrants. Specifically, the findings suggest that (a) Filipino American immigrants experienced ethnic and cultural denigration in the Philippines prior to their U.S. arrival, (b) ethnic and cultural denigration in the Philippines and in the United States may lead to the development of colonial mentality (CM), and (c) that CM may have negative mental health consequences among Filipino American immigrants. The two studies' findings suggest that the Filipino American immigration experience cannot be completely captured by the voluntary immigrant narrative, as they provide empirical support to the notion that the Filipino American immigration experience needs to be understood in the context of colonialism and its most insidious psychological legacy- CM.


Provisional stenting in the real world: results in 1058 consecutive patients undergoing percutaneous coronary angioplasty.
OBJECTIVE: To study a strategy of aggressive coronary balloon angioplasty with provisional stenting in allcomers. In randomized trials, stenting has improved the outcome of patients undergoing coronary intervention. However, whether these results hold up in clinical practice is largely unknown. Furthermore, the results of balloon angioplasty have also improved dramatically. It is therefore essential to evaluate the current results of balloon angioplasty and to assess whether stents are required in all patients. METHODS: The authors prospectively studied the occurrence of death, myocardial infarction (MI) and target lesion revascularization (TLR) of a large consecutive group of patients undergoing aggressive balloon angioplasty with provisional stenting. None of the patients received a platelet glycoprotein IIb/IIIa receptor blocker. The results were compared with the outcome of routine stenting in recent randomized trials. RESULTS: Angioplasty was performed in 1058 patients of whom 369 (34.9%) received a stent. The angiographic success rate was 98.9%. During hospital stay, 4.8% of the patients suffered any cardiac event. At one-year follow-up, death occurred in 1.1%, MI in 3.3%, TLR in 12.4% and any event in 16.7% of the patients. Event-free survival at one-year was 82.3%. These results compare favorably with routine stenting in recent trials. CONCLUSIONS: Aggressive balloon angioplasty with provisional stenting yields excellent results in a general patient population.


Identification of a tumor-associated target antigen, ATM-1, for a human T-cell clone with activated killer activity and its existence in sera of cancer patients.
Three human T-cell clones with activated killer activity (5B5, 5C1, and 7B5) which could lyse various tumor cell lines were established. The cytotoxic activity of these clones was decreased by incubation with anti-CD3 monoclonal antibody, suggesting that they recognized tumor cells by T-cell antigen receptor. A monoclonal antibody which blocked the cytotoxic activity of clone 5B5 was obtained. This antibody (N1977) blocked the binding and cytotoxic activity of clone 5B5 at the target cell level, suggesting that the antigen defined by N1977 antibody, designated as ATM-1, was a target molecule recognized by 5B5 cells. ATM-1 in the conditioned medium of a cancer cell line (NBT-2) and serum from a patient with lung cancer was characterized by following its immunoreactivity. On gel filtration, both the conditioned medium and the serum gave three peaks of ATM-1 immunoreactivity, corresponding to approximate molecular weights of 1,200,000, 700,000, and 120,000, respectively. They were chromatofocused at pH 4.0, 4.8, and 6.5, respectively. The high molecular weight forms were shown to be molecules with the disulfide-linked elementary glycoprotein with ATM-1 immunoreactivity and approximate molecular weight of 120,000. Most of the molecules with ATM-1 immunoreactivity bound to both concanavalin A and wheat germ agglutinin, and their binding activity to the antibodies was lost by treatment at 60 degrees C for 30 min. An assay of ATM-1 level in sera was performed by a sandwich enzyme immunoassay. The following positive percentages were obtained from preliminary clinical studies: breast cancer, 67% (8 of 12 cases); hepatocellular carcinoma, 83% (10 of 12 cases); gastric cancer, 58% (7 of 12 cases); lung cancer, 41% (5 of 12 cases); hematological malignancies, 0% (0 of 9 cases); systemic lupus erythematosus, 0% (0 of 8 cases); rheumatoid arthritis, 0% (0 of 8 cases).


Numerical approximation of tangent moduli for finite element implementations of nonlinear hyperelastic material models.
Finite element (FE) implementations of nearly incompressible material models often employ decoupled numerical treatments of the dilatational and deviatoric parts of the deformation gradient. This treatment allows the dilatational stiffness to be handled separately to alleviate ill conditioning of the tangent stiffness matrix. However, this can lead to complex formulations of the material tangent moduli that can be difficult to implement or may require custom FE codes, thus limiting their general use. Here we present an approach, based on work by Miehe (Miehe, 1996, "Numerical Computation of Algorithmic (Consistent) Tangent Moduli in Large Strain Computational Inelasticity," Comput. Methods Appl. Mech. Eng., 134, pp. 223-240), for an efficient numerical approximation of the tangent moduli that can be easily implemented within commercial FE codes. By perturbing the deformation gradient, the material tangent moduli from the Jaumann rate of the Kirchhoff stress are accurately approximated by a forward difference of the associated Kirchhoff stresses. The merit of this approach is that it produces a concise mathematical formulation that is not dependent on any particular material model. Consequently, once the approximation method is coded in a subroutine, it can be used for other hyperelastic material models with no modification. The implementation and accuracy of this approach is first demonstrated with a simple neo-Hookean material. Subsequently, a fiber-reinforced structural model is applied to analyze the pressure-diameter curve during blood vessel inflation. Implementation of this approach will facilitate the incorporation of novel hyperelastic material models for a soft tissue behavior into commercial FE software.


[Rhabdomyosarcoma in middle to old-aged patients: analysis of clinicopathological features and prognosis in 76 cases].
Albeit rare, rhabdomyosarcoma (RMS) may occur in middle to old-aged adults. The purpose of this study was to investigate the clinicopathological characteristics and prognosis of RMS in patients ≥ 40 years of age. The clinical, pathological and immunohistochemical profiles of the 76 cases were reviewed, and the outcomes were analyzed. There were 46 males and 30 females with a median age of 55 years. Sixty cases occurred in somatic soft tissues, including extremities (n = 25), head and neck (n = 19) and trunk (n = 16). Sixteen developed in the internal organs with predilection for the urogenital tract. Histologically, 38 cases (50.0%) were of embryonal RMS (ERMS) subtype, 29 cases (38.2%) of pleomorphic RMS (PRMS), 7 cases (9.2%) of poorly-differentiated RMS (PdRMS), and 2 cases (2.6%) of alveolar RMS (ARMS). Immunohistiochemically, the tumor cells showed diffuse staining of desmin (68/68, 100%), with a variable expression of myogenin (30/43, 69.8%), MyoD1 (33/51, 64.7%) and MSA (29/40, 72.5%). The therapy data available in 54 cases showed that three inoperable cases were treated with chemotherapy and radiotherapy, 51 cases underwent local excision, including 10 cases with chemotherapy, 9 cases with radiotherapy, 8 cases with chemotherapy and radiotherapy after surgery, and the remaining 24 cases were not administrered with postoperative adjuvant therapy. The follow-up data available in 54 cases showed that 22 patients were alive with unresectable or recurrent/disease and 32 patients were alive with no evidence of disease. The median disease-free and overall survivals were 6.0 months and 7.0 months, respectively. Twenty-seven patients exhibited progression of disease with recurrence in 8 cases and metastasis in 19 cases. The median interval to progression was 6.0 months. The most common subtypes of RMS in middle to old aged patients are ERMS and PRMS. In general, Rhabdomyosarcomas of middle to old aged patients shows a highly aggressive behavior. Patients may benefit from multi-disciplinary therapy.


Promising Gene Therapeutics for Salivary Gland Radiotoxicity.
More than 0.5 million new cases of head and neck cancer are diagnosed worldwide each year, and approximately 75% of them are treated with radiation alone or in combination with other cancer treatments. A majority of patients treated with radiotherapy develop significant oral off-target effects because of the unavoidable irradiation of normal tissues. Salivary glands that lie within treatment fields are often irreparably damaged and a decline in function manifests as dry mouth or xerostomia. Limited ability of the salivary glands to regenerate lost acinar cells makes radiation-induced loss of function a chronic problem that affects the quality of life of the patients well beyond the completion of radiotherapy. The restoration of saliva production after irradiation has been a daunting challenge, and this review provides an overview of promising gene therapeutics that either improve the gland's ability to survive radiation insult, or alternately, restore fluid flow after radiation. The salient features and shortcomings of each approach are discussed.


Derisking Drug-Induced Carcinogenicity for Novel Therapeutics.
Assessing the carcinogenic potential of innovative drugs spanning diverse therapeutic modalities and target biology represents a major challenge during drug development. Novel modalities, such as cell and gene therapies that involve intrinsic genetic modification of the host genome, require distinct approaches for identification of cancer hazard. We emphasize the need for customized weight-of-evidence cancer risk assessments based on mode of action that balance multiple options for preclinical identification of cancer hazard with appropriate labeling of clinical products and risk management plans. We review how advances in molecular carcinogenesis can enhance mechanistic interpretation and preclinical indicators of neoplasia, and recommend that drug targets be systematically assessed for potential association with tumorigenic phenotypes via genetic models and cancer genome resources.


Importin beta negatively regulates nuclear membrane fusion and nuclear pore complex assembly.
Assembly of a eukaryotic nucleus involves three distinct events: membrane recruitment, fusion to form a double nuclear membrane, and nuclear pore complex (NPC) assembly. We report that importin beta negatively regulates two of these events, membrane fusion and NPC assembly. When excess importin beta is added to a full Xenopus nuclear reconstitution reaction, vesicles are recruited to chromatin but their fusion is blocked. The importin beta down-regulation of membrane fusion is Ran-GTP reversible. Indeed, excess RanGTP (RanQ69L) alone stimulates excessive membrane fusion, leading to intranuclear membrane tubules and cytoplasmic annulate lamellae-like structures. We propose that a precise balance of importin beta to Ran is required to create a correct double nuclear membrane and simultaneously to repress undesirable fusion events. Interestingly, truncated importin beta 45-462 allows membrane fusion but produces nuclei lacking any NPCs. This reveals distinct importin beta-regulation of NPC assembly. Excess full-length importin beta and beta 45-462 act similarly when added to prefused nuclear intermediates, i.e., both block NPC assembly. The importin beta NPC block, which maps downstream of GTPgammaS and BAPTA-sensitive steps in NPC assembly, is reversible by cytosol. Remarkably, it is not reversible by 25 microM RanGTP, a concentration that easily reverses fusion inhibition. This report, using a full reconstitution system and natural chromatin substrates, significantly expands the repertoire of importin beta. Its roles now encompass negative regulation of two of the major events of nuclear assembly: membrane fusion and NPC assembly.


Anti-cancer effect of oncolytic adenovirus-armed shRNA targeting MYCN gene on doxorubicin-resistant neuroblastoma cells.
Chemotherapy is one of the few effective choices for patients with neuroblastoma. However, the development of muti-drug resistance (MDR) to chemotherapy is a major obstacle to the effective treatment of advanced or recurrent neuroblastoma. The muti-drug resistance-associated protein (MRP), which encodes a transmembrane glycoprotein, is a key regulator of MDR. The expression of MRP is a close correlation with MYCN oncogene in neuroblastoma. We have recently shown ZD55-shMYCN (oncolytic virus armed with shRNA against MYCN) can down-regulate MYCN to inhibit tumor cells proliferation and induce apoptosis in neuroblastoma. Here we further report ZD55-shMYCN re-sensitized doxorubicin-resistant cells to doxorubicin (as shown by reduced proliferation, increased apoptosis, and inhibited cell migration), and reduced the in vivo growth rate of neuroblastoma xenografts by down-regulation of MRP expression. Sequential therapy with doxorubicin did not affect the replication of ZD55-shMYCN in doxorubicin-resistant neuroblastoma cells, but decreased the expression of Bcl-2, Bcl-XL, MMP-1. Thus, this synergistic effect of ZD55-shMYCN in combination with doxorubicin provides a novel therapy strategy for doxorubicin-resistant neuroblastoma, and is a promising approach for further clinical development.


Pathways of sulfide oxidation by haloalkaliphilic bacteria in limited-oxygen gas lift bioreactors.
Physicochemical processes, such as the Lo-cat and Amine-Claus process, are commonly used to remove hydrogen sulfide from hydrocarbon gas streams such as landfill gas, natural gas, and synthesis gas. Biodesulfurization offers environmental advantages, but still requires optimization and more insight in the reaction pathways and kinetics. We carried out experiments with gas lift bioreactors inoculated with haloalkaliphilic sulfide-oxidizing bacteria. At oxygen-limiting levels, that is, below an O(2)/H(2)S mole ratio of 1, sulfide was oxidized to elemental sulfur and sulfate. We propose that the bacteria reduce NAD(+) without direct transfer of electrons to oxygen and that this is most likely the main route for oxidizing sulfide to elemental sulfur which is subsequently oxidized to sulfate in oxygen-limited bioreactors. We call this pathway the limited oxygen route (LOR). Biomass growth under these conditions is significantly lower than at higher oxygen levels. These findings emphasize the importance of accurate process control. This work also identifies a need for studies exploring similar pathways in other sulfide oxidizers such as Thiobacillus bacteria.


[The effect of electromagnetic stimulation on the depressive syndrome].
The authors describe the antidepressive effect of stimulation by static electromagnetical field of low induction, its value being 12.5 mT. They document the occurrence of the antidepressive effect by the results of their study on three groups of patients with the depressive syndrome, who were hospitalized in a mental hospital. 45 patients of the first group were subdued to a single stimulation of static electromagnetical field of 12.5 mT induction value, in the time schedule of 3x3 minutes with one minute breaks between the periods of electromagnetical stimulation. In 55% of them, the antidepressive effect has been observed, although of short duration. The experiments in the second group were arranged as being double blind. Patients in the stimulated group reacted by positive antidepressive effect more often than in the placebo group; the level of significance was 5%. In the third group of 19 patients, the stimulation was applied repeatedly. There was no evidence of confirmation of the assumption that repetition of stimulation could possibly stabilize and prolong its antidepressive effect, particularly by applying our used induction values. In spite of these facts it is possible to assume that in future the described antidepressive effect, after finding the appropriate manner, will be of use in therapeutical practice. (Tab. 4, Fig. 1, Ref. 7.)


Variation in the divalent cation requirements of influenza a virus N2 neuraminidases.
Influenza virus N2 neuraminidases were chromatographically purified from several vaccine candidate strains from 1957 to 1994. Enzymatic kinetic parameters and immunogenicity were tested for each strain. For each NA tested, with ionic strength held constant, Ca(2+) or Mg(2+) increased the initial rate of enzymatic activity. Earlier N2-NA strains had the highest initial velocity, V(max)/K(m) and V(max). There were significant differences among the influenza virus strains in enzymatic activity before and after addition of Ca(2+) or Mg(2+): V(max)/K(m) varied from 0.54 M(-1) s(-1) to 0.88 M(-1) s(-1) and V(max) varied from 2.45 s(-1) to 4.3 s(-1) before the addition of a divalent cation; and increased approximately 2-fold each of these kinetic parameters for each strain after the addition of exogenous Ca(2+) or Mg(2+). Exhaustive dialysis with EDTA reduced the initial velocity of each strain with significant differences found among strains, with a range of 0.1% to 8% of original activity. Activity was partially restored by the addition of exogenous Ca(2+) or Mg(2+), varying from 8% to 60% of pre-dialysis levels, but original rates were not achieved. This reduction in enzymatic activity for the tested strains (i.e., A/Japan/57 and A/Johannesburg/94) was accompanied by a parallel decrease in NA-immunogenicity, with antibody response decreasing by as much as 76% as measured by NI titer, and ELISA titer decreasing by as much as 68%. The addition of Ca(2+) or Mg(2+) to the post-dialysis sample restored immunogenicity to as much as 80% of pre-dialysis NI titers and as much as 78% of pre-dialysis ELISA titers. Dialysis had the least effect on early strains as measured by enzymatic kinetic parameters and immunogenicity studies. Zn(2+) had a slight inhibitory effect on the activity of all tested strains. Review of the nucleic acid sequence of each of these strains could not predict their enzymatic activity, immunogenicity or response to dialysis. If immunity against neuraminidase is desirable in vaccination against influenza, selection of vaccine candidate strains must include not only analysis of antigenic changes and sequence analysis but also enzymatic studies and determination of the requirement of divalent cations to maintain immunogenicity and activity during production.


World Wide Web resources for cancer support groups.
World Wide Web (internet) resources that pertain to cancer support groups, especially pain and symptom control, are listed and reviewed. How to start using these services; services for patients, families, and friends; disease-specific websites; and other types of electronic information repositories are discussed.


Takayasu arteritis: protean radiologic manifestations and diagnosis.
Diagnosis of Takayasu arteritis is difficult because the clinical features are similar to those of other diseases. In early-phase Takayasu arteritis, computed tomography (CT) and magnetic resonance (MR) imaging show thickening of the aortic wall. Late-phase Takayasu arteritis has been classified into four types: classic pulseless disease (type I), a mixed type (type II), the atypical coarctation type (type III), and the dilated type (type IV). In late-phase Takayasu arteritis, angiography usually demonstrates luminal changes such as stenosis, occlusion, or aneurysmal dilatation of the aorta and pulmonary artery and their branches. However, absence of such luminal changes does not exclude the possibility of early-phase Takayasu arteritis. Improvement in the clinical findings and subsidence of the active inflammatory process can be expected with early steroid treatment. Familiarity with the varied chest radiographic, angiographic, CT, and MR imaging features of Takayasu arteritis will permit earlier diagnosis and treatment.


Impulse response analysis of baroreceptor sensitivity.
The impulse response function (IRF) can express the dynamic relationship between systolic arterial pressure (SAP) and pulse interval (PI) and, consequently, represents an alternative method to assess baroreceptor sensitivity (BRS) in humans. Five normotensive and 13 hypertensive subjects (age 68 +/- 5 yr, range 60-74 yr) were studied at rest in the supine position during baseline conditions and after injections of phenylephrine and sodium nitroprusside. SAP and PI signals were derived from multiple 5-min noninvasive recordings of arterial blood pressure (Finapres) and electrocardiogram. Standard estimates of BRS were obtained by the slopes of transient changes in SAP and PI after the injection of phenylephrine and sodium nitroprusside (BRS(PE) and BRS(SNP)) and by spectral analysis (alpha-index). Impulse responses were obtained by the inverse Fourier transform of the transfer function between PI and SAP. The temporal pattern of the IRF was characterized by a main peak at t = 0, preceded by a "trough" at t = -1 s. A mathematical model of the baroreflex suggests that the peak value of IRF is linearly related to the BRS. The peak value and its smoothed version were shown to be significantly correlated to alpha, BRS(PE), and BRS(SNP) and significantly reduced in the hypertensive group during the three stages of the protocol. We suggest that IRF might be the ideal method to assess BRS because it does not require any subjective preselection of data segments or spectral bands.


Significance of SAA1.3 allele genotype in Japanese patients with amyloidosis secondary to rheumatoid arthritis.
To clarify the clinical significance of the SAA1.3 allele in the development and outcome of AA amyloidosis in Japanese patients with rheumatoid arthritis (RA). One hundred and twenty RA patients (60 alive and 60 dead) fulfilling the 1987 ACR criteria and 62 RA patients with biopsy-confirmed amyloid A (AA) amyloidosis (36 alive and 26 dead) were enrolled. The SAA1 genotypes were determined by PCR-based restriction fragment length polymorphism. To predict the clinical outcome of AA amyloidosis, we investigated characteristics and survival, focusing on the SAA1.3 allele retrospectively. The SAA1.3 allele genotype was not only a risk factor for the association of AA amyloidosis but also a poor prognostic factor for the development of AA amyloidosis (P=0.015). Both the association of AA amyloidosis arising early in the RA disease course and symptomatic variety and severity were found in amyloidotic patients with the SAA1.3 allele. The presenting factors adversely influenced were age (P=0.001), lowered serum albumin (P=0.001) and creatinine concentration (P=2.14 x 10(-5)). Renal involvement was associated with poor survival in patients with AA amyloidosis (P=0.011) and the presence of cardiac involvement was likely to be a risk factor for survival (P=0.062). The rate of the causes of death in respect to the category of infection, gastrointestinal diseases, and renal failure was higher in patients with AA amyloidosis than in those without amyloidosis, gastrointestinal diseases and renal failure. Cyclophosphamide was found to be superior to methotrexate in the management of RA patients with AA amyloidosis. Our data support the fact that homozygosity for the SAA1.3 allele is a univariate predictor of survival in addition to a risk factor for the association of AA amyloidosis adversely influencing the outcome in Japanese RA patients. Renal involvement is a pivotal clinical manifestation in the development of AA amyloidosis, as is likely to be cardiac involvement in AA amyloidosis secondary to RA.


The financial impact of ambulance diversions and patient elopements.
Admission process delays and other throughput inefficiencies are a leading cause of emergency department (ED) overcrowding, ambulance diversion, and patient elopements. Hospital capacity constraints reduce the number of treatment beds available to provide revenue-generating patient services. The objective of this study was to develop a practical method for quantifying the revenues that are potentially lost as a result of patient elopements and ambulance diversion. Historical data from 62,588 patient visits to the ED of a 450-bed nonprofit community teaching hospital in central Pennsylvania between July 2004 and June 2005 were used to estimate the value of potential patient visits foregone as a result of ambulance diversion and patients leaving the ED without treatment. The study hospital may have lost 3,881,506 dollars in net revenue as a result of ambulance diversions and patient elopements from the ED during a 12-month period. Significant revenue may be foregone as a result of throughput delays that prevent the ED from utilizing its existing bed capacity for additional patient visits.


Optimal control law for classical and multiconjugate adaptive optics.
Classical adaptive optics (AO) is now a widespread technique for high-resolution imaging with astronomical ground-based telescopes. It generally uses simple and efficient control algorithms. Multiconjugate adaptive optics (MCAO) is a more recent and very promising technique that should extend the corrected field of view. This technique has not yet been experimentally validated, but simulations already show its high potential. The importance for MCAO of an optimal reconstruction using turbulence spatial statistics has already been demonstrated through open-loop simulations. We propose an optimal closed-loop control law that accounts for both spatial and temporal statistics. The prior information on the turbulence, as well as on the wave-front sensing noise, is expressed in a state-space model. The optimal phase estimation is then given by a Kalman filter. The equations describing the system are given and the underlying assumptions explained. The control law is then derived. The gain brought by this approach is demonstrated through MCAO numerical simulations representative of astronomical observation on a 8-m-class telescope in the near infrared. We also discuss the application of this control approach to classical AO. Even in classical AO, the technique could be relevant especially for future extreme AO systems.


Sinogenic polymicrobial anaerobic orbital cellulitis: the importance of source control and broad empirical antibiotics.
A man in his twenties with a history of recurrent sinusitis was urgently referred to the emergency department (ED) by an out-of-hours general practitioner following a 2-day history of increasing right eye pain, redness and swelling after a week of coryzal symptoms. He denied visual impairment and any history of recent dental pain or procedures. Initial assessment in ED noted fever, tachycardia and hypotension. Video consultation with ophthalmologist in the ED identified proptosis, periorbital erythema and chemosis with full eye movement solely affecting the right eye. Visual acuity of 6/6 was confirmed in both eyes. After review by the ear, nose and throat (ENT) team, a diagnosis of sinogenic right orbital cellulitis was made, empirical antibiotics started and care transferred to the ENT team for immediate surgical intervention. 48 hours postoperatively, the patient acutely deteriorated, developing ophthalmoplegia and visual acuity of 6/95 in the right eye. Repeat imaging demonstrated a deteriorating picture and urgent surgery was organised at a neighbouring hospital's specialist ENT unit combined with a change to his antibiotics. On day 4, 1 day following transfer, an anaerobic bacterium, Eggerthia catenaformis, was isolated from blood cultures collected on admission. The patient improved clinically following the second surgery and targeted antimicrobial therapy, eventually being discharged 10 days after initial presentation. In addition to E. catenaformis, the Anaerobic Reference Unit (Cardiff) identified two further anaerobic bacteria, Parvimonas micra and Dialister pneumosintes This paper presents the first documented case of polymicrobial anaerobic orbital cellulitis secondary to acute bacterial sinusitis. Moreover, this case underpins the importance of broad empirical antibiotics coupled with surgical source control to effectively manage a rare but sight-threatening and life-threatening disease.


High data-rate communication link supported through the exploitation of optical channels in a characterized turbulent underwater environment.
Underwater turbulence presents a myriad of challenges for underwater optical systems through wavefront distortion and beam deflection. In this work, an underwater turbulence emulator is developed and thoroughly characterized to experimentally test the proposed underwater turbulence mitigation technique. This technique applies a modified HOBBIT system introduced in atmospheric turbulence to the relatively unknown underwater turbulence domain. By varying a beam's spatial position and relative phase gradient, a volume of turbulence is rapidly probed to determine the beam state for optimal propagation. This probe and control method is applied in multiple facets, including improved optical power transmission as well as supporting a 25-Gbps communication link through a dynamic environment.


Factors predicting poor outcome in patients with fulminant Guillaine-Barré syndrome.
This paper describes three patients with acute fulminant Guillain-Barrι Syndrome (GBS) with electrophysiologically inexcitable peripheral nerves not responding to two courses of intravenous immunoglobulin. Their clinical profile is compared with two other GBS patients having similar severity of disease but with demyelinative features, managed similarly during the same period. Patients who failed to respond were elderly with a mean age of 60 years, had prodromal diarrhea, rapid progression of muscle weakness requiring mechanical ventilation within 24 hours, dense weakness of all four limbs with cardiovascular autonomic symptoms and inexcitable peripheral nerves. The remaining two who recovered well were relatively younger with a mean age of 50 years, had no prodromal diarrhea, required ventilatory support by fourth day of illness, no cardiovascular autonomic symptoms and demyelinative neuropathy.


Physicochemical basis for formation of renal stones of calcium phosphate origin: calculation of the degree of saturation of urine with respect to brushite.
Brushite (CaHPO(4).2H(2)O) was considered to govern the formation of renal calculus of calcium phosphate origin. The degree of saturation of urine with respect to this phase was therefore calculated. This value was obtained from the ratio of the activity product of Ca(++) and HPO(4) (m) (K(sp)) before and after incubation of urine with brushite. The errors in the calculation of K(sp) were largely eliminated by this procedure.The urine of patients with idiopathic hypercalciuria and recurrent calcium-containing renal calculi was supersaturated with respect to brushit largely because of the high urinary concentration of Ca(++). The urine of normocalciuric subjects was undersaturated except at high urinary pH. This technique of estimating the degree of saturation of urine should allow a quantitative assessment of the various therapeutic regimens recommended for patients with nephrolithiasis.


Bioavailability of ascorbic acid in orange juice.
Several orange juice preparations were analyzed for biologically active and inactive ascorbic acid derivatives --L-ascorbic acid , dehydro-L-ascorbic acid, and inactive 2,3-diketo-L-gulonic acid. Fresh orange juice contained from 4 to 7 mg/100 ml of inactive ascorbic acid. In contrast, pasteurized orange juice contained from 15 to 30 mg/100 ml of inactive ascorbic acid (up to 400% more than fresh orange juice). Samples of fresh orange juice contained as much as 100% more active ascorbic acid than samples of pasteurized orange juice. There is great variability in prepared orange juice derivatives with a substantial decrease in bioavailability of active ascorbic acid.


In vivo antibacterial activity of MRX-I, a new oxazolidinone.
MRX-I is a potent oxazolidinone antibiotic against Gram-positive pathogens, including methicillin-resistant Staphylococcus aureus (MRSA), penicillin-resistant Streptococcus pneumoniae (PRSP), penicillin-intermediate S. pneumoniae (PISP), and vancomycin-resistant enterococci (VRE). In this study, the in vivo efficacy of orally administered MRX-I was evaluated using linezolid as a comparator. MRX-I showed the same or better efficacy than linezolid in both systemic and local infection models against the tested strains.


Localization of the gene for congenital dyserythropoietic anemia type I to a <1-cM interval on chromosome 15q15.1-15.3.
Congenital dyserythropoietic anemias (CDA) are a rare group of red-blood-cell disorders of unknown etiology that are characterized by ineffective erythropoiesis, pathognomonic cytopathology of the nucleated red blood cells in the bone marrow, and secondary hemochromatosis. In CDA type I, bone-marrow electron microscopy reveals characteristic findings in erythroid precursors, including spongy heterochromatin and enlarged nuclear pores. Since the genetic basis of CDA type I is not evident, we used homozygosity and linkage mapping to localize the genetic defect responsible for CDA type I in 25 Bedouins from four large consanguineous families. We report the linkage of this disease to markers on chromosome 15 located at q15. 1-q15.3. Fourteen markers within a 12-cM interval were typed in the relevant family members. Nine of the markers yielded maximum LOD scores of 1.625-12.928 at a recombination fraction of .00. Linkage disequilibrium was found only with marker D15S779. Haplotype analysis revealed eight different carrier haplotypes and highlighted the existence of a founder haplotype. Identification of historical crossover events further narrowed the gene location to between D15S779 and D15S778. The data suggest localization of the CDA type I gene within a 0.5-cM interval. The founder mutation probably occurred >/= 400 years ago. Sequence analysis of the coding region of protein 4.2, the only known erythroid-specific gene in the locus, did not reveal any change in the CDA type I patients. Future analysis of this locus may lead to the identification of a gene essential to normal erythropoiesis.


Seasonality effects on pharmaceuticals and s-triazine herbicides in wastewater effluent and surface water from the Canadian side of the upper Detroit River.
The influence of seasonal changes in water conditions and parameters on several major pharmacologically active compounds (PhACs) and s-triazine herbicides was assessed in the wastewater and sewage treatment plant (WSTP) effluent as well as the downstream surface water from sites on the Canadian side of the upper Detroit River, between the Little River WSTP and near the water intake of a major drinking water treatment facility for the City of Windsor (ON, Canada). The assessed PhACs were of neutral (carbamazepine, cotinine, caffeine, cyclophosphamide, fluoxetine, norfluoxetine, pentoxifylline, and trimethoprim) and acidic (ibuprofen, bezafibrate, clofibric acid, diclofenac, fenoprofen, gemfibrozil, indomethacin, naproxen, and ketoprofen) varieties. The major assessed s-triazine herbicides were atrazine, simazine, propazine, prometon, ametryn, prometryn, and terbutryn. At sampling times from September 2002 to June 2003, 15 PhACs were detected in the WSTP effluent at concentrations ranging from 1.7 to 1244 ng/L. The PhAC concentrations decreased by as much 92 to 100% at the Little River/Detroit River confluence because of the river dilution effect, with further continual decreases at sites downstream from the WSTP. The only quantifiable s-triazine in WSTP effluent, atrazine, ranged from 6.7 to 200 ng/L and was higher in Detroit River surface waters than in WSTP effluent. Only carbamazepine, cotinine, and atrazine were detectable at the low-nanogram and subnanogram levels in surface waters near a drinking water intake site. Unlike the PhACs, atrazine in the Detroit River is not attributable to point sources, and it is heavily influenced by seasonal agricultural usage and runoff. Detroit River surface water concentrations of carbamazepine, cotinine, and atrazine may present a health concern to aquatic wildlife and to humans via the consumption of drinking water.


Uterine rupture after previous low segment transverse cesarean is rarely catastrophic.
The cornerstone of concerns over trial of labor after cesarean (TOLAC) is the risk of uterine rupture. The purpose of this study was to document the rate of uterine rupture during TOLAC and to delineate its severity and consequences. We retrospectively collected the data on vaginal and cesarean deliveries after a previous cesarean section with specific emphasis on uterine rupture and dehiscence in our center from 2006 through 2013. 22,670 deliveries were registered, with 18.2% rate of cesarean section. 2890 women had a single cesarean scar; of them 1206 delivered vaginally and 194 were re-operated during unsuccessful TOLAC. Seven cases of uterine rupture and 16 cases of dehiscence were recorded. There were no maternal, intrapartum or neonatal deaths, and no cesarean hysterectomy. There was one re-laparotomy, one ICU admission, and one blood transfusion; one neonate was admitted to NICU. TOLAC was successful in 86.1% of cases. Cautious selection and close monitoring of candidates are the cornerstones of successful management of TOLAC. Readily available facilities for emergency cesarean delivery and concerted obstetrical team can save the mother and child from catastrophic complications.


[Laser trabeculoplasty--a review].
Laser therapy in open angle glaucomas represents an important step in their management, a topic of intense research during the second part of the XXth century. The initial enthousiasm generated by spectacular results obtained with minimal risks and complications, was soon tampered by their short survival. Nowadays, laser therapy is considered a second or third line therapy, alternative to surgery or intermediate stage between medication and surgery. There are selected cases in which laser therapy may be first line treatment in open angle glaucomas.


Molecular design for enhancement of ocular penetration.
Over the past two decades, many oral drugs have been designed in consideration of physicochemical properties to attain optimal pharmacokinetic properties. This strategy significantly reduced attrition in drug development owing to inadequate pharmacokinetics during the last decade. On the other hand, most ophthalmic drugs are generated from reformulation of other therapeutic dosage forms. Therefore, the modification of formulations has been used mainly as the approach to improve ocular pharmacokinetics. However, to maximize ocular pharmacokinetic properties, a specific molecular design for ocular drug is preferable. Passive diffusion of drugs across the cornea membranes requires appropriate lipophilicity and aqueous solubility. Improvement of such physicochemical properties has been achieved by structure optimization or prodrug approaches. This review discusses the current knowledge about ophthalmic drugs adapted from systemic drugs and molecular design for ocular drugs. I propose the approaches for molecular design to obtain the optimal ocular penetration into anterior segment based on published studies to date.


Solution Focused Brief Therapy in Post-Stroke Aphasia (SOFIA): feasibility and acceptability results of a feasibility randomised wait-list controlled trial.
The Solution Focused Brief Therapy in Post-Stroke Aphasia feasibility trial had four primary aims: to assess (1) acceptability of the intervention to people with aphasia, including severe aphasia, (2) feasibility of recruitment and retention, (3) acceptability of research procedures and outcome measures, and (4) feasibility of delivering the intervention by speech and language therapists. Two-group randomised controlled feasibility trial with wait-list design, blinded outcome assessors and nested qualitative research. Participants identified via two community NHS Speech and Language Therapy London services and through community routes (eg, voluntary-sector stroke groups). People with aphasia at least 6 months post stroke. Solution-focused brief therapy, a psychological intervention, adapted to be linguistically accessible. Participants offered up to six sessions over 3 months, either immediately postrandomisation or after a delay of 6 months. Primary endpoints related to feasibility and acceptability. Clinical outcomes were collected at baseline, 3 and 6 months postrandomisation, and at 9 months (wait-list group only). The candidate primary outcome measure was the Warwick-Edinburgh Mental Well-being Scale. Participants and therapists also took part in in-depth interviews. Thirty-two participants were recruited, including 43.8% with severe aphasia. Acceptability endpoints: therapy was perceived as valuable and acceptable by both participants (n=30 interviews) and therapists (n=3 interviews); 93.8% of participants had ≥2 therapy sessions (90.6% had 6/6 sessions). Feasibility endpoints: recruitment target was reached within the prespecified 13-month recruitment window; 82.1% of eligible participants consented; 96.9% were followed up at 6 months; missing data <0.01%. All five prespecified feasibility progression criteria were met. The high retention and adherence rates, alongside the qualitative data, suggest the study design was feasible and therapy approach acceptable even to people with severe aphasia. These results indicate a definitive randomised controlled trial of the intervention would be feasible. NCT03245060.


Adjuvant therapies utilization pattern and survival outcomes in high-grade head and neck soft tissue sarcoma; a population based study.
Head and neck soft tissue sarcoma (HNSTS) is a distinct entity within the soft tissue sarcoma group and the evidence supporting treatment options for this disease is poorly defined. Using the National Cancer Database, this study aims to reveal adjuvant therapy utilization patterns and their outcomes. HNSTS patients constituted 6.5% of all sarcoma patients and the analysis was limited to non-metastatic patients who underwent resection between 2004 and 2012. Chi-square test assessed the distribution of demographic, tumor and treatment variables in the treatment subgroups. Kaplan-Meier method estimated overall survival and proportional hazards model estimated survival hazard ratios for prognostic factors including age, comorbidity, gender, race, size, grade, margin status, histology, chemotherapy administration and radiotherapy technique/dose. The cohort included 2493 patients with a median age of 62years. Adjuvant therapies were more frequently delivered in high-grade, large tumors and with positive surgical margins. Limiting the analysis to 788 high-grade HNSTS patients, adjunctive radiotherapy improved 5-year survival from 44% (95% confidence interval (CI): 38-50) to 49% (CI: 43-55); reducing mortality hazards ratio by 24% (p<0.001). On multivariate analysis, older age, positive surgical margins and large tumor size were significant predictors of poor survival. These findings were consistent after propensity score analysis. Our analysis suggests that adjuvant radiotherapy is associated with improved survival in high-grade HNSTS. Higher dose and precise technique did not accentuate this benefit; however, further studies are needed to refine the treatment strategies, including the role of chemotherapy in the management of this rare disease.


Silencing Cenp-F weakens centromeric cohesion, prevents chromosome alignment and activates the spindle checkpoint.
Cenp-F is an unusual kinetochore protein in that it localizes to the nuclear matrix in interphase and the nuclear envelope at the G2/M transition; it is farnesylated and rapidly degraded after mitosis. We have recently shown that farnesylation of Cenp-F is required for G2/M progression, its localization to kinetochores, and its degradation. However, the role Cenp-F plays in mitosis has remained enigmatic. Here we show that, following repression of Cenp-F by RNA interference (RNAi), the processes of metaphase chromosome alignment, anaphase chromosome segregation and cytokinesis all fail. Although kinetochores attach to microtubules in Cenp-F-deficient cells, the oscillatory movements that normally occur following K-fibre formation are severely dampened. Consistently, inter-kinetochore distances are reduced. In addition, merotelic associations are observed, suggesting that whereas kinetochores can attach microtubules in the absence of Cenp-F, resolving inappropriate interactions is inhibited. Repression of Cenp-F does not appear to compromise the spindle checkpoint. Rather, the chromosome alignment defect induced by Cenp-F RNA interference is accompanied by a prolonged mitosis, indicating checkpoint activation. Indeed, the prolonged mitosis induced by Cenp-F RNAi is dependent on the spindle checkpoint kinase BubR1. Surprisingly, chromosomes in Cenp-F-deficient cells frequently show a premature loss of chromatid cohesion. Thus, in addition to regulating kinetochore-microtubule interactions, Cenp-F might be required to protect centromeric cohesion prior to anaphase commitment. Intriguingly, whereas most of the sister-less kinetochores cluster near the spindle poles, some align at the spindle equator, possibly through merotelic or lateral orientations.


Protection by GABA and succinic semialdehyde of seed germination and some enzymatic activities against high concentration of hydroxylamine.
Hydroxylamine was found to stimulate germination of Lupinus albus at concentrations inferior to 10 mM and to inhibit it greatly at 20 mM concentration. This inhibition was partially restored by GABA or succinic semialdehyde. Hydroxylamine, at high concentrations, behaved as inhibitor in vivo on GABA 2-oxyglutarate amino-transferase and succinic semialdehyde dehydrogenase NAD-dependent, whereas it behaved as activator on succinic semialdehyde dehydrogenase NADP-dependent. No effects were observed on the enzymatic activities and the inhibited germination was partially restored, after GABA and succinic semialdehyde had been added to a growth medium with a 20 mM hydroxylamine concentration. A possible protection mechanism of GABA and succinic semialdehyde against hydroxylamine action is discussed.


A Bottom-up Synthesis of Antiaromatic Expanded Phthalocyanines: Pentabenzotriazasmaragdyrins, i.e. Norcorroles of Superphthalocyanines.
The first example of an antiaromatic expanded phthalocyanine, classified as a norcorrole of a superphthalocyanine has been prepared and fully characterized. The newly developed phthalonitrile dimerization reaction was a crucial step, which allowed for the bottom-up synthesis of expanded phthalocyanines. Their structure was confirmed by single crystal X-ray diffraction analysis. The 20 π antiaromaticity of the macrocycles was suggested by optical and theoretical calculations.


Serum tumor necrosis factor alpha, interleukin 1-beta, p24 antigen concentrations and CD4+ cells at various stages of human immunodeficiency virus 1 infection in children.
We measured the serum concentrations of tumor necrosis factor (TNF-alpha), interleukin 1-beta (IL-1-beta), p24 antigen, CD4+/CD8+ cells and immunoglobulins in 35 children at various stages of human immunodeficiency virus infection. Serum TNF-alpha concentrations were significantly higher in children with lymphocytic interstitial pneumonitis and in children with mildly symptomatic illness than in asymptomatic children or children with acquired immunodeficiency syndrome. In addition serum IL-1 concentrations were significantly higher in patients with lymphocytic interstitial pneumonitis than in asymptomatic, mildly symptomatic, or acquired immunodeficiency syndrome patients. Children with lymphocytic interstitial pneumonitis had the highest serum TNF-alpha and IL-1 concentrations. Among symptomatic children serum TNF-alpha concentrations correlated positively with those of IL-1, and both were inversely related to the amount of p24 antigen. TNF-alpha values in excess of 50 pg/ml were observed more frequently among patients with CD4+ cell count greater than 400/mm3 than in those with CD4+ cell count less than 400/mm3. We did not find any association between elevated TNF-alpha concentrations and cachexia, opportunistic infections or progressive encephalopathy.


Hexagonal SiC with spatially separated active sites on polar and nonpolar facets achieving enhanced hydrogen production from photocatalytic water reduction.
Sufficient spatial separation of photo-generated electrons and holes plays a significant role in affecting the efficiency for solar energy conversion. Non-equivalent facets of a catalyst are known to possess different charge distribution properties. Here, we report that hexagonal 6H-SiC, a metal-free, environmentally friendly, polar semiconductor, exhibits different charge distribution and photocatalytic properties on naturally occurring Si-{0001} and {10-10} facets. Very strong selectivity of metals in situ photodeposition occurs in these two facets, demonstrating that the photo-excited electrons are assembled only on polar Si-{0001} facets while the holes are assembled on non-polar {10-10} facets. Consequently, reduction reactions occur only on the Si-{0001} facets with noble metals, and meantime oxidation occurs only in {10-10} with metal oxide. We show that the activity of photocatalytic water splitting is significantly enhanced by this kind of selective depositions resulting from the charge spatial separation. The underlying mechanism is investigated in terms of experimental evidence and first principles calculations. Our results demonstrate that the utilization of facets with opposite catalytic characteristics could be a feasible means to enhance the photocatalytic performance in diverse semiconducting materials. This is, in particular, of interest for polar semiconductors, as their particles always naturally occur in both polar facets and non-polar ones without needing facet engineering.


Mixed systems of hydrophobically modified polyelectrolytes: controlling rheology by charge and hydrophobe stoichiometry and interaction strength.
Rheology and phase separation were investigated for aqueous mixtures of two oppositely charged hydrophobically modified polyelectrolytes. The typical phase separation, normally seen for oppositely charged polymer mixtures, is dramatically reduced by the presence of hydrophobic modification, and phase separation is only detected close to the point of charge neutralization. While the two polyelectrolytes separately can give high viscosities and a gel-like behavior, a pronounced maximum in viscosity and storage modulus with the mixing ratio of the polyelectrolytes is observed; the maximum is located between the points of charge and hydrophobe stoichiometry and reflects a combination of hydrophobic and electrostatic association. Lowering the charge density of the anionic polymer leads to a strengthened association at first, but at lower charge densities there is a weakened association due to the onset of phase separation. The strength of the electrostatic interaction was modified by adding salt. Increased ionic strength can lead to phase separation and to increased or decreased viscosity depending on the polyelectrolyte mixing ratio.


The molecular spectrum and clinical impact of DIS3 mutations in multiple myeloma.
Multiple myeloma (MM) is a plasma cell neoplasm that presents with a major biological and clinical heterogeneity. We here investigated the spectrum of clonal and subclonal mutations of DIS3, an active part of the exosome complex, that may play a role in the development or progression of MM. The whole coding sequence of DIS3 was subjected to deep sequencing in 81 uniformly-treated MM patients and 12 MM cell lines and the overall occurrence of DIS3 mutations as well as the presence of DIS3 mutations in minor and major subclones were correlated with cytogenetic alterations and clinical parameters. Our study identified DIS3 mutations in 9/81 patients that were associated with 13q14 deletions and IGH translocations on the cytogenetic level. Specifically, we detected seven novel somatic DIS3 single nucleotide variants (SNVs) and defined three hot spot mutations within the RNB domain. Lastly, we found a trend towards a shorter median overall survival for patients with DIS3 mutations, and patients carrying DIS3 mutations in minor subclones of their tumours showed a significantly worse response to therapy compared to patients with DIS3 mutations in the major subclone.


A Comparison of the Selectivity of Extraction of [PtCl6](2-) by Mono-, Bi-, and Tripodal Receptors That Address Its Outer Coordination Sphere.
Extraction and binding studies of [PtCl6](2-) are reported for 24 mono-, bi-, and tripodal extractants containing tris(2-aminoethyl)amine (TREN) or tris(3-aminopropyl)amine (TRPN) scaffolds. These reagents are designed to recognize the outer coordination sphere of [PtCl6](2-) and to show selectivity over chloride anion under acidic conditions. Extraction from 0.6 M HCl involves protonation of the N-center in tertiary amines containing one, two, or three urea, amide, or sulfonamide hydrogen-bond donors to set up the following equilibrium: 2L(org) + 2H(+) + [PtCl6](2-) ⇌ [(LH)2PtCl6](org). All reagents show higher Pt loading than trioctylamine, which was used as a positive control to represent commercial trialkylamine reagents. The loading of [PtCl6](2-) depends on the number of pendant amides in the extractant and follows the order tripodal > bipodal > monopodal, with urea-containing extractants outperforming amide and sulfonamide analogues. A different series of reagents in which one, two, or three of the alkyl groups in tris-2-ethylhexylamine are replaced by 3-N'-hexylpropanamide groups all show a comparably high affinity for [PtCl6](2-) and high selectivity over chloride anion in extractions from aqueous acidic solutions. (1)H NMR titration of three extractants [LH·Cl] with [(Oct4N)2PtCl6] in CDCl3 provides evidence for high selectivity for [PtCl6](2-) over chloride for tri- and bipodal extractants, which show higher binding constants than a monopodal analogue.


Mental health of vulnerable groups experiencing a drought or bushfire: A systematic review.
Natural hazards are increasing because of climate change, and they disproportionately affect vulnerable populations. Prior reviews of the mental health consequences of natural hazard events have not focused on the particular experiences of vulnerable groups. Based on the expected increase in fires and droughts in the coming years, the aim of this systematic review is to synthesize the global evidence about the mental health of vulnerable populations after experiencing natural hazards. We searched databases such as Ovid MEDLINE, EMBASE, CINAHL and Ovid PsycInfo using a systematic strategy, which yielded 3,401 publications. We identified 18 eligible studies conducted in five different countries with 15,959 participants. The most common vulnerabilities were living in a rural area, occupying a low socioeconomic position, being a member of an ethnic minority and having a medical condition. Common experiences reported by vulnerable individuals affected by drought included worry, hopelessness, isolation and suicidal thoughts and behaviors. Those affected by fire reported experiencing posttraumatic stress disorder (PTSD) and anger. These mental health problems exacerbated existing health and socioeconomic challenges. The evidence base about mental health in vulnerable communities affected by natural hazards can be improved by including standardized measures and comparison groups, examining the role of intersectional vulnerabilities, and disaggregating data routinely to allow for analyses of the particular experiences of vulnerable communities. Such efforts will help ensure that programs are informed by an understanding of the unique needs of these communities.


5'-NUCLEOTIDASES OF NAJA NAJA KARACHIENSIS SNAKE VENOM: THEIR DETERMINATION, TOXICITIES AND REMEDIAL APPROACH BY NATURAL INHIBITORS (MEDICINAL PLANTS).
Present study was carried out regarding enzymatic assay for 5'-nucleotidase enzymes present in snake venom Naja naja karachiensis and to evaluate twenty eight medicinal plants as their antidotes. Elevated enzymatic activities i.e., 119, 183, 262 and 335 U/mL were observed in 10, 20, 30 and 40 µg of crude venom, respectively, in dose dependent manner. Among various plant extracts only two (Bauhinia vaiiegate L. and Citms linion (L.) Burm. f.) were found 94% effective at 160 µg to neutralize 112 U/mL activities (p 0.5) while reference standard was proved 93.2% useful at 80 pg to halt 111 U/mL activities. Cedrus deodara G. Don, Enicostemna hyssopifolium (Willd.) Verdoom, Terminalia arjuma Wight & Am. and Zingiber officinalis Rosc. (at 160 µg) were found ≥90% effective (0.5 ≥ p ≥ 0.1) while Citrulus colocynthis, Fogonia cretica L., Rhazya stticta Dcne and Stenolobiun stans (L.) D. Don (at 320 µg) were proved 90% effective (0.05 ≥ p ≥ 0.02). The remaining plant extracts were observed abortive (p ≥ 0.001) in neutralization of 5'-nucleotidases enzymatic actions. This study emphasizes further characterization of active plant extracts to further explore the antivenom influences of these herbal remedies against deleterious effects produced by 5'-nucleotidase enzymes after snake bite envenomation.


Intact Auditory Cortical Cross-Frequency Coupling in Early and Chronic Schizophrenia.
Previous work has identified a hierarchical organization of neural oscillations that supports performance of complex cognitive and perceptual tasks, and can be indexed with phase-amplitude coupling (PAC) between low- and high-frequency oscillations. Our aim was to employ enhanced source localization afforded by magnetoencephalography (MEG) to expand on earlier reports of intact auditory cortical PAC in schizophrenia and to investigate how PAC may evolve over the early and chronic phases of the illness. Individuals with early schizophrenia (n=12) (≤5 years of illness duration), chronic schizophrenia (n=16) (>5 years of illness duration) and healthy comparators (n = 17) performed the auditory steady state response (ASSR) to 40, 30, and 20 Hz stimuli during MEG recordings. We estimated amplitude and PAC on the MEG ASSR source localized to the auditory cortices. Gamma amplitude during 40-Hz ASSR exhibited a significant group by hemisphere interaction, with both patient groups showing reduced right hemisphere amplitude and no overall lateralization in contrast to the right hemisphere lateralization demonstrated in controls. We found significant PAC in the right auditory cortex during the 40-Hz entrainment condition relative to baseline, however, PAC did not differ significantly between groups. In the current study, we demonstrated an apparent sparing of ASSR-related PAC across phases of the illness, in contrast with impaired cortical gamma oscillation amplitudes. The distinction between our PAC and evoked ASSR findings supports the notion of separate but interacting circuits for the generation and maintenance of sensory gamma oscillations. The apparent sparing of PAC in both early and chronic schizophrenia patients could imply that the neuropathology of schizophrenia differentially affects these mechanisms across different stages of the disease. Future studies should investigate the distinction between PAC during passive tasks and more cognitively demanding task such as working memory so that we can begin to understand the influence of schizophrenia neuropathology on the larger framework for modulating neurocomputational capacity.


[Violence and mental health issues among Mexican adolescents that have considered or attempted cross-border migration].
The aim of this study was to estimate the role of victimization by violence among Mexican adolescents that have considered or attempted migrating to the United States, including mental health variables (emotional self-esteem, self-esteem in school, depression, suicidal ideation, and attempted suicide) as mediators of the effects. The study used a cross-sectional design with a stratified cluster sample of 13,198 adolescents from the 2nd Mexican National Survey on Exclusion, Intolerance, and Violence in public schools in 2009. The analysis used the regression models proposed by Baron & Kenny. Prevalence of having considered or attempted cross-border migration was 23.1%. Mean age was 16.36 years. Female adolescents constituted 54.9% of the sample, and 56% were lower-income. Mental health variables that acted as partial mediators were suicidal ideation (35.9%), depression (19.2%), attempted suicide (17.7%), emotional self-esteem (6.2%), and self-esteem in school (3.4%) for moderate family violence, and emotional self-esteem (17.5%) for social rejection in school and suicidal ideation (8.1%) for physical harm in school. Female adolescents showed greater impact from mediators than men in considering or having attempted cross-border migration. The study discusses the importance of incorporating the prevention of violence in the social contexts studied here and incorporating mental health in dealing with violence in adolescents and in public health programs in transit areas for illegal migrants.


Revised absolute stereochemistry of rhodiolosides A-D, rhodiolol A and sachalinol A from Rhodiola rosea.
The absolute stereochemistry of rhodiolosides A-D (1-4), four monoterpene glycosides from the roots of Rhodiola rosea, and their aglycones, rhodiolol A (5), (-)-rosiridol (6) and sachalinol A (7), were reinvestigated. It was found that the absolute configurations of C-4 in these compounds, previously assigned to be 4-R, should be revised to 4-S.


EGb761 provides a protective effect against Aβ1-42 oligomer-induced cell damage and blood-brain barrier disruption in an in vitro bEnd.3 endothelial model.
Alzheimer's disease (AD) is the most common form of senile dementia which is characterized by abnormal amyloid beta (Aβ) accumulation and deposition in brain parenchyma and cerebral capillaries, and leads to blood-brain barrier (BBB) disruption. Despite great progress in understanding the etiology of AD, the underlying pathogenic mechanism of BBB damage is still unclear, and no effective treatment has been devised. The standard Ginkgo biloba extract EGb761 has been widely used as a potential cognitive enhancer for the treatment of AD. However, the cellular mechanism underlying the effect remain to be clarified. In this study, we employed an immortalized endothelial cell line (bEnd.3) and incubation of Aβ(1-42) oligomer, to mimic a monolayer BBB model under conditions found in the AD brain. We investigated the effect of EGb761 on BBB and found that Aβ1-42 oligomer-induced cell injury, apoptosis, and generation of intracellular reactive oxygen species (ROS), were attenuated by treatment with EGb761. Moreover, treatment of the cells with EGb761 decreased BBB permeability and increased tight junction scaffold protein levels including ZO-1, Claudin-5 and Occludin. We also found that the Aβ(1-42) oligomer-induced upregulation of the receptor for advanced glycation end-products (RAGE), which mediates Aβ cytotoxicity and plays an essential role in AD progression, was significantly decreased by treatment with EGb761. To our knowledge, we provide the first direct in vitro evidence of an effect of EGb761 on the brain endothelium exposed to Aβ(1-42) oligomer, and on the expression of tight junction (TJ) scaffold proteins and RAGE. Our results provide a new insight into a possible mechanism of action of EGb761. This study provides a rational basis for the therapeutic application of EGb761 in the treatment of AD.


[Hyperlipidemia - side-effect of "Atypical" neuroleptics?].
After the introduction of the so-called 'atypical antipsychotics' some reports concerning hyperlipidemia observed in patients treated with these drugs have been published. The studies and case reports available up to now were reviewed. The available data show that hyperlipidemia particularly occurred in patients receiving clozapine, olanzapine and also quetiapine. Predominately elevated serum levels of triglycerides have been reported. The underlying pathomechanism still remains widely unclear. Since hyperlipidemia is an important symptom of the so called 'metabolic syndrome' which is often associated with severe complications like cardial and vascular diseases, more attention should be paid to hyperlipidemia as a potential side effect of antipsychotics.


Structure of organic-inorganic nanohybrids incorporating titanium(IV) oxoalkoxyacylate nanoclusters: a SANS study.
The small angle neutron scattering (SANS) technique was used to investigate the structure of nanohybrids consisting of a poly(methylmethacrylate) (PMMA) and one of two types of titanium(IV) nanoclusters. Cluster 1, [Ti6O4](OC2H5)8(CH2=CCH3COO)8, with polymerizable MMA ligands, formed covalent bonds with the polymer chains during the copolymerization, whereas cluster 2, [Ti6O4](OC2H5)8(CH3COO)8, had no polymerizable linkers and was blended into the polymeric matrix purely as a filler. In this study, SANS with contrast variation was used to investigate the size, shape and aggregation of the clusters in the hybrid materials, and their effect on the structure of the matrix. A polydispersed core-diffusion zone model was employed to explain the scattering contribution from the titanium clusters in both nanohybrid materials. No significant differences between the structures of the two nanohybrids were found. The fitted models suggest that the interface region between the cluster and matrix (the diffusion zone) is heavily occupied by the PMMA chains; however, they do not penetrate into the core region (titanium cluster).


Heat-dependent opening of TRPV1 in the presence of capsaicin.
Transient receptor potential vanilloid member 1 (TRPV1) is a Ca2+-permeable cation channel that serves as the primary heat and capsaicin sensor in humans. Using cryo-EM, we have determined the structures of apo and capsaicin-bound full-length rat TRPV1 reconstituted into lipid nanodiscs over a range of temperatures. This has allowed us to visualize the noxious heat-induced opening of TRPV1 in the presence of capsaicin. Notably, noxious heat-dependent TRPV1 opening comprises stepwise conformational transitions. Global conformational changes across multiple subdomains of TRPV1 are followed by the rearrangement of the outer pore, leading to gate opening. Solvent-accessible surface area analyses and functional studies suggest that a subset of residues form an interaction network that is directly involved in heat sensing. Our study provides a glimpse of the molecular principles underlying noxious physical and chemical stimuli sensing by TRPV1, which can be extended to other thermal sensing ion channels.


Endogenously produced Indian Hedgehog regulates TGFβ-driven chondrogenesis of human bone marrow stromal/stem cells.
Human bone marrow stromal/stem cells (hBMSCs) have an inherent tendency to undergo hypertrophy when induced into the chondrogenic lineage using transforming growth factor-beta 1 (TGFβ) in vitro, reminiscent of what occurs during endochondral ossification. Surprisingly, Indian Hedgehog (IHH) has received little attention for its role during hBMSC chondrogenesis despite being considered a master regulator of endochondral ossification. In this study, we investigated the role that endogenously produced IHH plays during hBMSC chondrogenesis. We began by analyzing the expression of IHH throughout differentiation using quantitative polymerase chain reaction and found that IHH expression was upregulated dramatically upon chondrogenic induction and peaked from days 9 to 12 of differentiation, which coincided with a concomitant increase in the expression of chondrogenesis- and hypertrophy-related markers, suggesting a potential role for endogenously produced IHH in driving hBMSC chondrogenesis. More importantly, pharmacological inhibition of Hedgehog signaling with cyclopamine or knockdown of IHH almost completely blocked TGFβ1-induced chondrogenesis in hBMSCs, demonstrating that endogenously produced IHH is necessary for hBMSC chondrogenesis. Furthermore, overexpression of IHH was sufficient to drive chondrogenic differentiation, even when TGFβ signaling was inhibited. Finally, stimulation with TGFβ1 induced a significant and sustained upregulation of IHH expression within 3 h that preceded an upregulation in all cartilage-related genes analyzed, and knockdown of IHH blocked the effects of TGFβ1 entirely, suggesting that the effects of TGFβ1 are being mediated through endogenously produced IHH. Together, our findings demonstrate that endogenously produced IHH is playing a critical role in regulating hBMSC chondrogenesis.


[Effect of highly active antiretroviral therapy on the clinical onset of AIDS defined by clinical episodes].
The frequency of opportunistic infections and the mortality of HIV-infected patients have changed notably since the widespread use of highly active antiretroviral therapy. Accordingly, the spectrum of first AIDS-defining conditions may have changed too. The aim of the present study was to analyze whether there have been changes in the clinical presentation of AIDS after the extensive use of highly active antiretroviral therapy. We performed an analysis of the presentation forms of AIDS in 190 patients with a prospective follow-up who developed this disease between April 1989 and December 2000. We compared the first AIDS-defining conditions occurring until December 1996 with those occurring after January 1997. Visceral leishmaniasis was the first AIDS-defining condition in 18 patients (14.2%) during the first period, whereas it was the AIDS-onset disease in two cases (3.1%) during the second period (P = 0.01). Tuberculosis was the first diagnosis in 63 patients (50%) in the first period and in 26 cases (40.6%) in the second period (P = 0.22). The frequency of other first AIDS-defining disorders remained unchanged along the whole study time. With regard to the clinical onset of AIDS, there has been a significant reduction in the frequency of visceral leishmaniasis as the first AIDS-defining disorder after the introduction of highly active antiretroviral therapy. Tuberculosis continues to be the more frequent clinical debut condition of AIDS.


Determination of nanogram level of VSCs using GC-SCD and an adsorption tube.
A quantitative analytical method was used for detection of low level of volatile sulfur compounds (VSCs) using GC with a sulfur chemiluminescence detector (SCD) in this study. A linear response over the range of 2-90 ngS injected was obtained with a good repeatability or reproducibility. Equimolar response for H2S, DMS, MeSH, and EtSH was obtained by use of SCD and the response of the SCD is nearly equimolar for different sulfur compounds. It was possible to quantify the total VSCs as well as individual VSC using one of the standard VSCs. VSC recovery was measured with respect to some storing methods. An adsorption tube packed with molecular sieve 5A showed almost perfect recovery for both H2S and dimethyl sulfide (DMS) during 6 days at a dark state below 2 degrees. Whereas, with a gas tight pyrex vial or tedlar bag, it was impossible to obtain the recovery of 50% after 6 days. It is strongly recommended to use the adsorption tube for determination of nanogram levels of volatile sulfur compounds and for storing and concentrating VSCs effectively with a minimum experimental error.


Three-dimensional computed tomographic reconstructions of intracranial meningiomas.
In a series of four patients with intracranial meningiomas, three-dimensional computed tomographic (CT) reconstructions were a useful diagnostic and surgical adjunct. Three-dimensional images are created from standard CT data by a boundary-detecting computer software program. Three-dimensional images of tumor invading or adjacent to the bony calvarium are projected about the x, y, and z axes. Axial and sagittal sections delineate the lesions. The images created allow a surgical view of the meningiomas in three-dimensional space and demonstrate the relationship of these masses to the skull. The use of three-dimensional reconstructions in craniofacial surgery and in neurosurgery is reviewed.


In vivo depletion of hematopoietic stem cells in the rat by an anti-CD45 (RT7) antibody.
Anti-CD45 monoclonal antibodies (mAbs) are potentially powerful tools for the depletion of mature leukocytes. As their application for immunotherapy also depends on their effects on bone marrow (BM) progeny, the in vivo effects of an anti-CD45 mAb (anti-RT7(a) mAb) on BM precursor cells were analyzed in a rat model. Anti-RT7(a) mAb treatment was performed in LEW.1W (RT1(u) RT7(a)) rats with the use of different dosages. In addition, major histocompatibility complex (MHC)-congenic BM transplantation making use of a diallelic polymorphism (RT7(a)/RT7(b)) of rat CD45 was applied. Following injection of anti-RT7(a) mAb into normal LEW.1W rats, T cells were profoundly depleted in blood, lymph nodes, and spleen, whereas B cells were coated only by the antibody. Single injection of anti-RT7(a) mAb in a high dose induced a lethal aplastic syndrome with severe thrombocytopenia. Rescue of antibody-treated animals with BM from congenic LEW.1W-7B rats (RT1(u) RT7(b)) and transplantation of BM from LEW.1W rats pretreated with anti-RT7(a) mAb into sublethally irradiated LEW.1W-7B recipients revealed a profound effect of the mAb on progeny of myeloid and T-cell lineage. Following repeated antibody treatment of stable mixed chimeras (RT7(b)/RT7(a)), very few RT7(a)-positive B cells were still detectable after 6 months and their number declined during the subsequent year. These observations show that this anti-RT7(a) mAb effectively depletes mature T cells as well as BM precursor cells of myeloid, T-cell, and thrombocytic lineage after in vivo application. In contrast, mature B cells are not depleted, but precursors also appear to be eliminated. Overall, the findings suggest that the anti-RT7(a) mAb efficiently depletes early rat hematopoietic stem cells.


Symptom Management and Lifestyle Modifications in Multiple Sclerosis.
This article reviews the rationale and approach to symptom management and lifestyle modifications in multiple sclerosis (MS). MS symptoms are important to treat because they affect quality of life and daily activity. Appreciation of cluster symptoms (where one symptom contributes to another), changes over time, and multimodality therapeutic approaches are guiding optimized symptom management. Equally important are lifestyle modifications that enhance central nervous system reserve and function. These modifications are the foundation for a health maintenance, wellness, and vascular risk factor control program. Symptom management and lifestyle modifications are important therapeutic targets to improve the lives of patients with MS.


Activation of protein serine/threonine phosphatase PP2Cα efficiently prevents liver fibrosis.
Over-activation of TGFβ signaling pathway and uncontrolled cell proliferation of hepatic stellate cells (HSCs) play pivotal roles in liver fibrogenesis, while the protein serine/threonine phosphatase PP2Cα was reported to negatively regulate TGFβ signaling pathway and cell cycle. Our study aimed to investigate the role of PP2Cα in liver fibrogenesis. The effects of PP2Cα activation on liver fibrosis were investigated in human HSCs and primary rat HSCs in vitro using western blotting, real-time PCR, nuclear translocation, cell viability and cell cycle analyses. The antifibrogenic effects in carbon tetrachloride (CCl(4))- and bile duct ligation (BDL)-induced mice in vivo were assessed using biochemical, histological and immunohistochemical analyses. The results demonstrated that activation of PP2Cα by overexpression or the new discovered small molecular activator NPLC0393 terminated TGFβ-Smad3 and TGFβ-p38 signaling pathways, induced cell cycle arrest in HSCs and decreased α-smooth muscle actin (α-SMA) expression, collagen deposition and hepatic hydroxyproline (HYP) level in CCl(4)- and BDL-induced mice. Our findings suggested that PP2Cα activation might be an attractive new strategy for treating liver fibrosis while the small molecular activator NPLC0393 might represent a lead compound for antifibrogenic drug development. Moreover, our study might provide the first evidence for the role of PP2C family members in the fibrotic disease.


The C4 cycle and beyond: diverse metabolic adaptations accompany dual-cell photosynthetic functions in Setaria.
C4 photosynthesis is typically characterized by the spatial compartmentalization of the photosynthetic reactions into mesophyll (M) and bundle sheath (BS) cells. Initial carbon fixation within M cells gives rise to C4 acids, which are transported to the BS cells. There, C4 acids are decarboxylated so that the resulting CO2 is incorporated into the Calvin cycle. This work is focused on the study of Setaria viridis, a C4 model plant, closely related to several major feed and bioenergy grasses. First, we performed the heterologous expression and biochemical characterization of Setaria isoforms for chloroplastic NADP-malic enzyme (NADP-ME) and mitochondrial NAD-malic enzyme (NAD-ME). The kinetic parameters obtained agree with a major role for NADP-ME in the decarboxylation of the C4 acid malate in the chloroplasts of BS cells. In addition, mitochondria-located NAD-ME showed regulatory properties that could be important in the context of the operation of the C4 carbon shuttle. Secondly, we compared the proteomes of M and BS compartments and found 825 differentially accumulated proteins that could support different metabolic scenarios. Most interestingly, we found evidence of metabolic strategies to insulate the C4 core avoiding the leakage of intermediates by either up-regulation or down-regulation of chloroplastic, mitochondrial, and peroxisomal proteins. Overall, the results presented in this work provide novel data concerning the complexity of C4 metabolism, uncovering future lines of research that will undoubtedly contribute to the expansion of knowledge on this topic.