Characterization of antibodies produced by S-s- individuals

BACKGROUND: Historically, classification of U- and U variant (U+var) individuals has been made by hemagglutination and adsorption and elution studies performed with polyclonal U antisera. Molecular studies and serologic tests with a potent monoclonal anti-He have shown that U+var red cells, some of which are He+, possess an altered form of glycophorin B. STUDY DESIGN AND METHODS: Seventeen sera, previously determined to contain anti-U, were tested with a panel of red cells of common and rare MNS types. RESULTS: Five sera contained anti-U only, and 12 sera contained broadly reactive antibodies with apparent, but inseparable, anti-U,He or anti-U,N,He specificities. CONCLUSION: The majority of antibodies produced by S-s-U- individuals are anti-U plus anti-glycophorin B and are analogous to the broadly reactive antibodies produced by En(a-) individuals whose red cells lack glycophorin A or have altered glycophorin A. To avoid further immunization of patients with anti-U, sera used for classification of S-s-U- donors should be selected to detect S-s- red cells that possess altered forms of glycophorin B.     

Characterization of four beta-lactamases produced by Staphylococcus aureus.

Staphylococcus aureus produces four types of beta-lactamase (A, B, C, and D). To investigate the effect of specific beta-lactamase type upon staphylococcal resistance, each beta-lactamase was purified to homogeneity, and the Michaelis constants (Km values) and turnover numbers (kcat values) for various penicillin and cephalosporin substrates were determined. Whereas Km values of the four beta-lactamases were comparable for penicillin G, cephalothin, and cefamandole, the type A and D enzymes exhibited greater affinity than the type B and C beta-lactamases for nitrocefin, cefazolin, and cephapirin. Conversely, the type B and C beta-lactamases exhibited greater kcat values than the type A and D enzymes against most of the cephalosporin agents, excluding nitrocefin. In contrast to earlier reports suggesting that the type B beta-lactamase is relatively inefficient in hydrolyzing penicillin G, we found only minor differences in the specific activities and kcat values of the type A, B, and C beta-lactamases. The type D beta-lactamase was distinctly less active against penicillin G, however, exhibiting only 15 to 25% of the kcat values of the other beta-lactamases. More than a 2,000-fold difference between the relative efficiencies of hydrolysis (kcat/Km) of cefazolin and cefuroxime by the type A beta-lactamase exists. This greatly exceeds the 60-fold difference in the stability of penicillin G and cefazolin with the same enzyme. Whereas the isoelectric points of the type A, B, and C beta-lactamases were similar, the value for the type D beta-lactamase was distinguishably lower (10.1 for types A, B, and C and 9.7 for type D).We conclude that marked differences in the stability of commonly used beta-lactams to hydrolysis by the staphylococcal beta-lactamases are present. This heterogeneity and the clinical implication thereof need to be considered in the antibiotic management of staphylococcal infection.     

Characterization of a pentylenetetrazol-like interoceptive stimulus produced by ethanol withdrawal

Rats were trained with food reinforcement to discriminate the anxiogenic drug pentylenetetrazol (PTZ, 20 mg/kg) from saline in a two-lever-choice task. In Experiment 1, ethanol, 8.25% w/v was given by gavage (7/day) for 4 days, with doses titrated to maintain moderate intoxication. After termination of ethanol, the rats exhibited mild overt signs of withdrawal and, in discrimination tests with saline as the test substance, they selected the PTZ lever, an effect reversed by ethanol, 2 g/kg, and by diazepam, 5 mg/kg. In Experiment 2, rats drank a nutritionally complete liquid diet containing ethanol, 4.5% w/v, for 1 week. They became tolerant to the intoxicating effect of ethanol, and blood ethanol concentration mounted with continued dosing. On termination of chronic ethanol, rats selected the PTZ lever before the onset of overt physical signs of withdrawal, and both measures returned to base line within 3 days. In Experiment 3 the percentage of rats selecting the PTZ lever after termination of ethanol depended upon the dose (up to 12.5 g/kg) and duration (up to a ceiling effect by 3 days) of ethanol administered chronically. These results indicate that a PTZ-like stimulus produced interoceptively can be demonstrated in the rat as an objective measure of ethanol withdrawal. This paradigm may provide insight into the symptom of anxiety associated with ethanol withdrawal.     

Characterization of displaced white matter by brain tumors using combined DTI and fMRI

In vivo white matter tractography by diffusion tensor imaging (DTI) has become a popular tool for investigation of white matter architecture in the normal brain. Despite some unresolved issues regarding the accuracy of DTI, recent studies applied DTI for delineating white matter organization in the vicinity of brain lesions and especially brain tumors. Apart from the intrinsic limitations of DTI, the tracking of fibers in the vicinity or within lesions is further complicated due to changes in diseased tissue such as elevated water content (edema), tissue compression and degeneration. These changes deform the architecture of the white matter and in some cases prevent definite selection of the seed region of interest (ROI) from which fiber tracking begins. We show here that for displaced fiber systems, the use of anatomical approach for seed ROI selection yields insufficient results. Alternatively, we propose to select the seed points based on functional MRI activations which constrain the subjective seed ROI selection. The results are demonstrated on two major fiber systems: the pyramidal tract and the superior longitudinal fasciculus that connect critical motor and language areas, respectively. The fMRI based seed ROI selection approach enabled a more comprehensive mapping of these fiber systems. Furthermore, this procedure enabled the characterization of displaced white matter using the eigenvalue decomposition of DTI. We show that along the compressed fiber system, the diffusivity parallel to the fiber increases, while that perpendicular to the fibers decreases, leading to an overall increase in the fractional anisotropy index reflecting the compression of the fiber bundle. We conclude that definition of the functional network of a subject with deformed white matter should be done carefully. With fMRI, one can more accurately define the seed ROI for DTI based tractography and to provide a more comprehensive, functionally related, white matter mapping, a very important tool used in pre-surgical mapping.     

Characterization and mode of action of a bacteriocin produced by a Bacteroides fragilis strain.

A Bacteroides fragilis strain produces a low-molecular-weight (13,500 to 18,700), proteinaceous bacteriocin during the stationary growth phase. The extracellular bacteriocin is not inducible by ultraviolet light or mitomycin C and is stable between pH 7.5 and 8.2. The majority of the bacteriocin is thermolabile, but a small proportion (3%) of the bacteriocin is stable after autoclaving at 121 degrees C for 15 min. Killing of sensitive bacteroides cells follows single-hit kinetics, and the interaction of a single molecule of bacteriocin with a target cell occurs in two stages. The killing of susceptible cells is affected by temperature and the growth state of the susceptible cells. The bacteriocin is unusual in that the primary event in its mode of action is the inhibition of RNA synthesis. The bacteriocin inhibits RNA synthesis immediately but has no effect on DNA synthesis or intracellular ATP levels. Protein synthesis is inhibited after a delay of 20 min, presumably as a result of the initial inhibition of RNA synthesis.     

Characterization of TEM-56, a novel beta-lactamase produced by a Klebsiella pneumoniae clinical isolate

TEM-56 produced by a Klebsiella pneumoniae clinical isolate is a novel beta-lactamase of isoelectric point 6.4 that confers a moderate resistance level to expanded-spectrum cephalosporins. The amino acid sequence deduced from the corresponding bla gene showed two amino acid replacements with respect to the TEM-2 sequence: Glu-104 to Lys and His-153 to Arg. This enzyme showed catalytic properties close to those of TEM-18. Thus, TEM-56 appears as a new TEM mutant, an intermediary between TEM-18 and the extended-spectrum beta-lactamase TEM-21.     

Characterization and biological activity against Naegleria fowleri of amoebicins produced by Bacillus licheniformis D-13.

The strain Bacillus licheniformis D-13 produces three hydrophobic peptides (amoebicins d13-A, d13-B, and d13-C) that elicit antiamoebic activity against human-pathogenic and nonpathogenic species of Naegleria and have a broad spectrum of antibacterial activity. The three amoebicins have the same amino acid composition (three Asp, two Glu, two Val, and nine Leu residues) and molecular weight (1,870). Amoebicin d13-B causes lysis of amoebae through disorganization of the cell membrane. It also induces permeability to 86Rb and membrane disruption in asolectin vesicles.     

Characterization of secondary hyperalgesia produced by topical capsaicin jelly--a new experimental tool for pain research.

Peripheral administration of the nociceptive agent capsaicin is used as an experimental tool to study neurophysiological and pharmacological aspects of the generation and control of pain. When investigating secondary hyperalgesia phenomena, current topical and intradermal capsaicin delivery methods have two key limitations. Intradermal injection can evoke severe chemogenic pain and both delivery methods produce an unstable area of dynamic mechanical allodynia. We present validity studies of a new preparation for capsaicin delivery that overcomes these limitations. The novel capsaicin formulation consists of a water-based semisolid jelly preparation containing 1% capsaicin which is applied topically under adhesive-free occlusion to a small area of the skin. We demonstrate that in healthy human subjects this model evokes areas of flare, punctate hyperalgesia and mechanical allodynia which are equivalent to established models and that these areas are stable over time and reproducible on repeat experiments. The jelly formulation evokes only minimal chemogenic pain and, as the preparation remains in situ throughout the study providing constant capsaicin exposure, a stable area of dynamic mechanical allodynia is produced. These validation studies show that this novel capsaicin administration method is a practical, reliable and viable tool for investigating experimental secondary hyperalgesia.     

人胰型及唾液型淀粉酶的分离纯化与鉴定

目的 为获得高比活性、高纯度的人胰型及唾液型淀粉酶。方法 用硫酸铵分级沉淀、Ｓｅ ｐｈａｄｅｘ Ｇ－５０凝胶层析及ＤＥＡＥ－纤维素离子交换层析法分别从人胰腺及唾液中分离纯化人胰型及唾液型淀粉酶,并用聚丙烯酰胺凝胶电泳法进行纯度、酶活性及分子量鉴定。结果 获得了高比活性及高纯度的人胰型及唾液型淀粉酶,胰型及唾液型淀粉酶的表现分子量分别为５５ＫＤ和５７ＫＤ。结论 本方法是获得高纯度及高比活性人胰型及唾     

Proteases produced by vibrios

Bacteria of the genus Vibrio are normal habitants of the aquatic environment but the some species are believed to be human pathogens. Pathogenic vibrios produce various pathogenic factors, and the proteases are also recognized to play pathogenic roles in the infection: the direct roles by digesting many kinds of host proteins or indirect roles by processing other pathogenic protein factors. Especially VVP from Vibrio vulnificus is thought to be a major pathogenic factor of the vibrio. Although HA/P, the V. cholerae hemagglutinin/protease, is not a direct toxic factor of cholera vibrio, its significance is an undeniable fact. Production of HA/P is regulated together with major pathogenic factors such as CT (cholera toxin) or TCP (toxin co-regulated pilus) by a quorum-sensing system. HA/P is necessary for full expression of pathogenicity of the vibrio by supporting growth and translocation in the digestive tract. Processing of protein toxins such as CT or El Tor hemolysin is also an important pathogenic role.     

Characterization of secondary hyperalgesia produced by topical capsaicin jelly—a new experimental tool for pain research

Peripheral administration of the nociceptive agent capsaicin is used as an experimental tool to study neurophysiological and pharmacological aspects of the generation and control of pain. When investigating secondary hyperalgesia phenomena, current topical and intradermal capsaicin delivery methods have two key limitations. Intradermal injection can evoke severe chemogenic pain and both delivery methods produce an unstable area of dynamic mechanical allodynia. We present validity studies of a new preparation for capsaicin delivery that overcomes these limitations. The novel capsaicin formulation consists of a water-based semisolid jelly preparation containing 1% capsaicin which is applied topically under adhesive-free occlusion to a small area of the skin. We demonstrate that in healthy human subjects this model evokes areas of flare, punctate hyperalgesia and mechanical allodynia which are equivalent to established models and that these areas are stable over time and reproducible on repeat experiments. The jelly formulation evokes only minimal chemogenic pain and, as the preparation remains in situ throughout the study providing constant capsaicin exposure, a stable area of dynamic mechanical allodynia is produced. These validation studies show that this novel capsaicin administration method is a practical, reliable and viable tool for investigating experimental secondary hyperalgesia.     

Characterization of a psoriatic skin model produced with involved or uninvolved cells

Current knowledge suggests that uninvolved psoriatic skin could demonstrate characteristics associated with both normal and involved psoriatic skins. However, the triggering factor allowing the conversion of uninvolved skin into a psoriatic plaque is not fully understood. To counter this lack of information, we decided to develop pathological skin substitutes produced with uninvolved psoriatic cells, in order to better characterize the uninvolved psoriatic skin. Substitutes were produced using the self‐assembly approach. Macroscopic, immunohistochemical, permeability and physicochemical results showed that involved substitutes had a thicker epidermis, higher cell proliferation, abnormal cell differentiation and a more permeable and disorganized stratum corneum compared with normal substitutes. Various results were observed using uninvolved cells, leading to two proposed profiles: profile 1 was suggested for uninvolved skin substitutes mimicking the results obtained with normal skin substitutes; and profile 2 was dedicated to those mimicking involved skin substitutes in all aspects that were analysed. In summary, uninvolved substitutes of profile 1 had a thin, well‐organized epidermis with normal cell proliferation and differentiation, such as observed with normal substitutes, while uninvolved substitutes of profile 2 showed an inverse trend, i.e. a thicker epidermis, higher cell proliferation, abnormal cell differentiation and a more disorganized and more permeable stratum corneum, such as seen with involved substitutes. The results suggest that uninvolved substitutes could demonstrate characteristics associated with both normal or involved psoriatic skins. Copyright © 2012 John Wiley & Sons, Ltd.     

Macroamylase immunoglobulins show high affinity for animal and human amylases

We have examined the affinity shown by the immunoglobulin fraction from each of five sera containing macroamylase for amylases from different sources: human saliva or human, porcine, or ovine pancreas. High affinity constants, 0.4 X 10(10) to 7.2 X 10(10) L/mol, were found in competitive binding experiments with human or porcine pancreatic amylase. All but one serum yielded linear Scatchard plots, indicating that in most sera the amylase-binding immunoglobulins are homogeneous, possibly monoclonal. The immunoglobulin fractions from different sera differed in their specificity: two of them bound all four types of amylases, whereas two bound only one type. Three of the five immunoglobulin fractions showed considerably higher affinity towards one or both of the animal amylases than towards the human ones, and may be primarily directed against some animal amylase.     

消化系神经内分泌肿瘤超声造影表现特征分析

目的分析消化系统神经内分泌瘤(NET)原发病灶及肝转移灶超声造影(CEUS)表现特征,评价CEUS诊断消化系NET的价值。方法 2005—2009年Charité医院收治90例经手术和(或)穿刺活检病理确诊的消化系NET患者。回顾性分析79个消化系NET原发病灶、510个肝转移灶常规超声表现及69个原发病灶、70个肝转移灶CEUS表现。结果常规超声示79个消化系NET原发病灶以低回声和混合回声为主,69.7%(23/33)的小肠NET原发病灶和64.3%(18/28)的胰腺NET原发病灶内部回声不均,27.8%(22/79)的消化系NET原发病灶含部分液化区。CEUS示69个消化系NET原发病灶中93.3%(63/69)血供丰富,72.5%(50/69)呈向心性增强。常规超声示510个肝转移灶中39.1%(199/510)表现为低回声,34.1%(174/510)为高回声,23.7%(121/510)为混合回声;32.7%(167/510)的肝转移灶周围存在声晕。CEUS示肝转移灶中70.0%(49/70)为向心性增强,27.1%(19/70)为快速整体增强,呈离心充盈的仅占2.9%(2/70);78.7%(55/70)的肝转移灶在60s内开始出现增强消退。结论消化系NET原发病灶及肝转移灶具有特征性CEUS表现。CEUS可作为NET影像诊断学方法之一。     

Characterization of the volatile organic compounds produced from green coffee in different years by gas chromatography ion mobility spectrometry

The effect of storage time on green coffee volatile organic compounds (VOCs) was studied by their separation via head space solid-phase microextraction and identification via gas chromatography-ion mobility spectrometry. In total, 38 kinds of VOCs, mainly composed of alcohols, aldehydes, esters and ketones, were identified. The fingerprint showed that the VOCs produced by green coffee in different years had obvious differences, especially, acrolein, 3-methylbutyl acetate, butanoic acid, heptan-3-ol, and so on, that could be used to predict the storage time. In addition, with the increase of storage time, the contents of butanal, ethanol, dimethyl sulfide, propanal, butan-2-one had no obvious change, and could be considered as typical aroma characteristics of green coffee or special aroma components for variety identification. Meanwhile, principal component analysis (PCA) and “nearest neighbor” fingerprint analysis could also effectively distinguish green coffee with different storage times. Comprehensive analysis showed that GC-IMS technology could provide strong and favorable support for coffee storage.

The effect of storage time on green coffee VOCs was studied by their separation via HS-SPME and identification via GC-MS.     

Characterization and diagnostic confidence of contrast-enhanced ultrasound for solid renal tumors

The objective was to determine whether contrast-enhanced ultrasound (CEUS) could improve the diagnostic confidence of solid renal masses. CEUS examinations were performed on 51 patients with renal tumors. Histologic findings from surgical specimens (n = 24) or magnetic resonance imaging follow-up (n = 27) were used as reference procedures for definitive diagnosis. Diffuse heterogeneous/homogeneous enhancement and quick peripheralnodularenhancement were found to be characteristic patterns in renal cell carcinoma (RCC). Dotlike or diffuse heterogeneous/homogeneous enhancement and slow peripheral nodular enhancement were observed as typical enhancement patterns in angiomyolipoma. The results show that CEUS combined with conventional ultrasound significantly improves diagnostic confidence. The sensitivity for RCC diagnosis with this imaging approach was 86% and the specificity was 93%. Both positive and negative predictive values of detection were 90% and the overall accuracy was 90%.     

Neovascularization produced by angiotensin II

Following vascular occlusion, development of collateral circulation occurs in at least two time-related phases: (1) the fast enhancement of the function of preexisting channels and (2) the slow formation of new vessels. Inasmuch as the renin-angiotensin system can act as a protective mechanism against local ischemia by activating preexisting collateral vessels, it is of interest to establish whether angiotensin II also produces stimulation of new vessel formation. Angiotensin II or cholecystokinin, an unrelated peptide, was incorporated in a slow-release formulation polyacrylamide gel and implanted in a pocket made in the rabbit cornea. Periodic examinations revealed that angiotensin II significantly stimulates new vessel formation; maximum values were attained in approximately 2 to 3 weeks. In contrast, cholecystokinin or polyacrylamide gel alone failed to stimulate any significant new vessel formation. Positive neovascularization was present in 85% of the total number of corneas implanted with angiotensin II, whereas 14% and 8% positive results were seen in the corneas implanted with either cholecystokinin or polyacrylamide gel alone, respectively. It is concluded that angiotensin II not only facilitates the activation of preexisting collateral vascular pathways but also has angiogenic properties and therefore could play an active role not only in the fast but also in the slow phase of the development of collateral circulation.