Hemiorchidectomy leads to dramatic and immediate alterations in pituitary follicle-stimulating hormone secretion and the functional activity of the remaining testis in the adult male bonnet monkey (Macaca radiata)

The aim of the present study was to examine the effect of hemiorchidectomy (HO) on serum FSH, LH, testosterone (T), and inhibin (INH) concentrations as well as on the testicular volume (TV) and on changes in the kinetics of germ cell turnovers in the remaining testis of adult male bonnet monkeys. Blood samples collected at 2200 h at various times before and after HO and testicular biopsies obtained at different periods were subjected to hormone analysis and DNA flow cytometry. Though serum T levels were lowered (p < 0.05) at 12 h after HO, T levels rapidly returned to intact control concentrations by Day 5. While serum LH remained unaltered, serum FSH increased markedly within 2 days of HO and remained significantly (p < 0.05) elevated over the next 90 days. Though serum INH showed a significant decrease (p < 0.05) by 15 min of HO, it returned to approximately 80% of intact levels within one week. The TV of the remaining testis showed maximal increment by Day 30 (p < 0.05) of HO. DNA flow cytometric analysis 24 days after HO showed increases (p < 0.05) in spermatogonia (2C) and primary spermatocytes (4C). These cell types by Day 45 had transformed to round (1C) and elongate (HC) (by 38%, p < 0.001) spermatids. Overall spermatogenesis (conversion of 2C to 1C and HC) showed significant enhancement at Days 110 and 175, suggesting that the spurt in spermatogenic activity is not confined to a single spermatogenic cycle.

Drug-drug co-crystals: Temperature-dependent proton mobility in the molecular complex of isoniazid with 4-aminosalicylic acid

Two drug-drug co-crystals of the anti-tuberculosis drugs isoniazid (INH), pyrazinamide (PYR) and 4-aminosalicylic acid (PAS) are reported. The first is the 1 : 1 molecular complex of INH and PAS. The second is the monohydrate of the 1 : 1 complex of PYR and PAS. The crystal structures of both co-crystals are characterized by a number of hydrogen bonded synthons. Hydrogen bonding of the COOH center dot center dot center dot N-pyridine type is found in both cases. In the INH : PAS co-crystal, there are two symmetry independent COOH center dot center dot center dot center dot N-pyridine hydrogen bonds. In one of these, the H-atom is located on the carboxylic group and is indicative of a co-crystal. In the second case, partial proton transfer occurs across the hydrogen bond, and the extent of proton transfer depends on the temperature. This is more indicative of a salt. Drug-drug co-crystals may have some bearing in the treatment of tuberculosis.

Del INI a la SEPI, la empresa pública en España durante la segunda mitad del siglo XX

Lan honekin historiako garai ezberdinek, batez ere XX. mendeko bigarren erdikoa, enpresa publikoaren gain izandako eragina ikusi nahi izan da. Horrez gain, sektore batzuetan enpresa publikoaren jarduera eta horietan lortu zituen emaitzak aztertu nahi izan dira. Beraz, helburua Espainiako enpresa publikoaren historia ikustea izan da. Horregatik, lehenengo enpresa publikoaren definizioa eta aurrekariak (XVIII. eta XIX. mendeak) azaldu dira eta, horren ondoren, XX. mendea aztertu da. Horretarako, XVIII. errege fabrikak eta pribilegiodun merkatal konpainia batzuk eta XIX. mendeko liberalismoak horien gain izandako eragina ikusi da. XX. mendeari dagokionez, ikusi denez, gutxienez bigarren erdian, INI-a izan zen garrantzia gehien izan zuen erakundea eta horregatik, bere jarduera eta politiken bilakaera aztertu da batez ere. Horrez gain, INI-a desagertu arte sortutako beste bi erakunde ikusi dira INH eta TENEO. Azkenik, INI-a ordezkatu zuten AIE eta SEPI erakundeak ikusi dira.

Perfil genético por sequenciamento e genotipagem de cepas multirresistentes do Mycobacterium tuberculosis provenientes de diversos estados brasileiros

A tuberculose multirresistente (MR) a drogas é uma séria ameaça à saúde pública devido à maiores complexidade, custo e efeitos colaterais do tratamento. Poucos estudos descreveram a epidemiologia molecular de isolados de Mycobacterium tuberculosis MR no Brasil. Neste trabalho foi investigada a diversidade genética e mutações associadas à resistência a drogas de 99 isolados MR e 7 não MR coletados em um período de 8 anos e provenientes de 12 estados brasileiros. Esta investigação foi feita através da análise do polimorfismo de fragmentos de restrição do elemento de inserção IS6110 (IS6110-RFLP), spoligotyping e sequenciamento de regiões dos genes rpoB e katG que conferem resistência aos antibióticos rifampicina e isoniazida, respectivamente. Mutações nos genes katG e rpoB foram encontradas em 90,9% e 93% dos isolados MR analisados, respectivamente. Para o gene rpoB, 91,9% das mutações estavam contidas na região RRDR de 81-pb. Um total de 51 (51.5%), 23 (23.3%) e 11 (11.1%) isolados MR apresentaram mutações nos códons 531, 526 e 516, respectivamente. Com relação ao gene katG, foram encontradas mutações em 93% dos isolados MR analisados, sendo que 7 apresentaram mutações apenas na primeira região analisada (katG1). O codon 315 da segunda região analisada do gene katG (katG2) apresentou mutações em 82.8% dos isolados MR, sendo a maioria Ser315Thr. A região katG1 apresentou mutações em 30.3% dos isolados MR sendo a maioria deleção do códon 4. Pelo spoligotyping foi possível determinar que os isolados MR deste estudo pertencem a 5 diferentes famílias (com suas subfamílias) de M. tuberculosis circulantes no Brasil, onde as mais frequentemente encontradas foram: LAM (46%), T (17%) e H (12%). Nós observamos que uma das famílias, a EAI5, carrega mutações no códon 463 do gene katG, o que não ocorre para as demais. Além disso, entre nossos isolados foi identificada um isolado pertencente à cepa Beijing (extremamente virulenta), mas este fato não é alarmante já que se tratou de apenas um caso. Através de nossos dados foram descritos novos alelos mutados para os genes rpoB e katG. Com exceção da família X2, foi identificada uma região inicial do gene katG com alta frequência de mutações nos isolados MR. A análise por IS6110-RFLP revelou que 25 isolados formaram 11 grupos genotípicos enquanto 74 mostraram um padrão único de bandas. Esta alta taxa de polimorfismo indica aquisição independente de resistência entre nossos isolados. Para a família H, foi identificada uma inversão na freqüência de ocorrência de mutações no gene rpoB, sendo o códon 516 o mais mutado, seguido pelo 526 e 531. Os resultados deste estudo fornecem informações úteis para um melhor entendimento do espectro de mutações dos isolados MR de pacientes no Brasil. Nossos resultados também se tornam úteis no desenvolvimento de testes diagnósticos de tuberculose MR e para auxiliar no rastreamento da transmissão global desta doença.

天然产物中的HIV-1 非核苷类逆转录酶抑制剂

综述了天然产物中对H IV 21 逆转录酶具有抑制活性的化合物的来源和分类。重点介绍了几种H IV 21 非核 苷类逆转录酶抑制剂(NNRT Is) 的性质、特点、抗H IV 21 活性及其构效关系。从天然资源和传统中草药发现具有新 结构、新作用机制的天然NNRT Is 是开发研制抗H IV 21 药物的一条极有希望的途径。

金环蛇毒心脏毒对S180,EAC腹水癌细胞的细胞毒性作用

目的: 探讨金环蛇毒心脏毒对S180, EAC 腹水癌细胞的细胞毒作用。方法: 采用小白鼠腹腔和皮下接种S180, EAC 腹 水癌细胞造成小白鼠腹水模型后腹腔注射金环蛇毒心脏毒。结果: 腹腔注射金环蛇毒心脏毒, 能抑制肿瘤细胞的生长, 降低接 种率。但不能完全控制腹水和癌细胞的生长。体外试验表明有明显的细胞毒作用。台酚蓝染色镜检可见死细胞显著增加, 腹 水图片检查, 给药后细胞膜破裂, 纤维化坏死明显。结论: 能延长小白鼠存活时间。

金环蛇蛇毒对S180, EAC 腹水癌细胞的细胞毒性作用

　目的　探讨金环蛇毒对S180, EAC 腹水癌细胞的细胞毒性作用。　方法　采用小白鼠腹腔和皮下 接种S180, EAC 腹水癌细胞造成小白鼠腹水模型后腹腔注射金环蛇毒。　结果　腹腔注射金环蛇毒, 能 抑制肿瘤细胞的生长, 降低接种率。但不能完全控制腹水和癌细胞的生长。体外试验表明有明显的细胞毒 作用。台酚蓝染色镜检可见死细胞显著增加, 腹水图片检查给药后细胞膜破裂, 纤维化坏死明显。　结论 　能延长小白鼠存活时间。

大蹼铃蟾新型抗菌肽的研究

两栖类动物皮肤裸露，柔软而湿润，体表富含腺体，其皮肤分泌物含有大量的具有特殊分子结构、功能复杂多样的生物活性分子。从两栖类己经分离到的包括血管紧张素样肤，促甲状腺素释放肤，蛙啡肤，舒缓激肤样多肤，速激肤样多肤，雨蛙肤样多肽等14类多肤家族以及生物胺生物碱等。"大蹼铃蟾是两栖类无尾目的一种，是我国的一个特有物种，主要分布于中国西南，其中云南山区种群数量巨大，多生活在污水糖和沟渠中，生活环境富有大量病原体。其皮肤腺体发达，受刺激能大量分泌富含活性物质的粘液，是分离生物活性分子的优良来源。其中抗菌肤作为活性分子的一大类，在大蹼铃蟾皮肤分泌物中不断有新的发现。通过大蹼铃蟾皮肤cDNA文库的构建，并对cDNA文库中的793个长度大于500个碱基的cDNA克隆进行随机测序，得到一类新的大蹼铃蟾抗菌肤，命名为Inaximins。由4个cDNA克隆推测得到这一类的新的抗菌肤，一共可以分为5个不同的种类，分别命名为maximinS1-5。这一类由cD瓜序列推测而来的新型抗菌肤，除了maximinS1只有14个氨基酸残基组成，maXiminS2-5都由18个氨基酸残基。并且从氨基酸残基序列上分析，新的maximinS与以往分离的到的大蹼铃蟾抗菌肤InaX1in和IllaXi"inH都不具有相似性。和东方铃蟾、多彩铃蟾皮肤分泌物中分离到的bobinin和bOInininH相比较，序列上也无同源性。从cDNA的结构上看，其编码的前体蛋白呈现一种全新的排列方式。每个cDNA克隆编码的maxininS前体蛋白都是由两个部分：信号肤加上一组不同maximinS和间隔肤的不同排列。这种奇特的排列组合方式，在东方铃蟾和多彩铃蟾都未曾发现。出现这个排列方式的原因，是由于基因本身结构，或是由于mRNA的可变剪切，或是由于体细胞基因重排，都不得而知，需要进一步的研究除了maximinS1，maximinS2-5，其二级结构分析都显示出一种类似的两亲性的Q螺旋空间结构。为了检测这一类新的大蹼铃蟾的活性，maximinsl和maximinS4用人工合成的方法得到，并对其进行了抗菌肚活性的测定。活性测定的结果显示，相同条件下，maximinS1不具抗菌活性，maximinS4具有显著的抗支原体的活性。这些现象间接证明了，小分子的抗菌肤其功能可能与氨基酸序列无关，而和空间构象由更加密切的联系。由于抗菌肤maximinS是由cDNA序列推导而来，并没有直接通过生化分离方法从大蹼铃蟾皮肤中得到。为了证明maximinS确实会被大蹼铃蟾表达，设计了一系列试验，将大蹼铃蟾皮肤分泌物通过简单的分子筛分离，追踪抗菌活性，并对具有活性的组分用MALDI-TOP质谱仪分析，证明maximinS类抗菌肤确实载大谱铃蟾皮肤分泌物中存在。

他克莫司对UVA照射后HaCaT细胞增殖的影响

　目的　探讨不同浓度他克莫司(TM)对不同强度长波紫外线(UVA)照射HaCaT细胞24h和48h后细胞 增殖活性的影响。方法将培养的HaCaT细胞分别行2, 4 和8J / cm2 的UVA 照射,且照射前1h 分别加入50, 500 和 5 000pg/mL的TM,对照组分别加入50, 500和5 000pg/mL的TM,但不行UVA照射。各剂量组分别照射24h和48h后在 倒置相差显微镜下观察细胞的形态变化,MTT法检测细胞的增殖活性。结果　HaCaT细胞经UVA照射24h后,细胞连接 松散,细胞折光性较未照射组差,部分细胞死亡、脱壁;与未经UVA照射组相比,细胞增殖受到抑制( P < 0. 05) ,加入TM 组无明显变化( P > 0. 05) ;照射48h后细胞虽有大量增殖,但体积较小;与未经UVA 照射组相比,细胞增殖明显( P < 0. 05) ,加入TM组细胞增殖受到抑制(P < 0. 05) 。结论　TM可抑制UVA照射HaCaT细胞引起的过度增殖,对UVA照射 后的HaCaT细胞有保护作用。

From PASS 1 to YES to AND logic: building parallel processing into molecular logic gates by sequential addition of receptors

The synthesis and photophysical characterization of a novel molecular logic gate 4, operating in water, is demonstrated based on the competition between. fluorescence and photoinduced electron transfer (PET). It is constructed according to a 'fluorophore-spacer-receptor(1)-spacer-receptor(2)' format where anthracene is the. fluorophore, receptor(1) is a tertiary amine and receptor(2) is a phenyliminodiacetate ligand. Using only protons and zinc cations as the chemical inputs and. fluorescence as the output, 4 is demonstrated to be both a two-input AND and INH logic gate. When 4 is examined in context to the YES logic gates 1 and 2, and the two-input AND logic gate 3 and three-input AND logic gate 5, each with one or more of the following receptors including a tertiary amine, phenyliminodiacetate or benzo-15-crown-5 ether, logic gate 4 is the missing link in the homologous series. Collectively, the molecular logic gates 1-5 corroborate the PET 'fluorophore-spacer-receptor' model using chemical inputs and a light-signal output and provide insight into controlling the. fluorescence quantum yield of future PET-based molecular logic gates.

A Coordination and Ligand Replacement Based Three-Input Colorimetric Logic Gate Sensing Platform for Melamine, Mercury Ions, and Cysteine

Discrimination of different species in various target scopes within a single sensing platform can provide many advantages such as simplicity, rapidness, and cost effectiveness. Here we design a three-input colorimetric logic gate based on the aggregation and anti-aggregation of gold nanoparticles (Au NPs) for the sensing of melamine, cysteine, and Hg2+. The concept takes advantages of the highly specific coordination and ligand replacement reactions between melamine, cysteine, Hg2+, and Au NPs. Different outputs are obtained with the combinational inputs in the logic gates, which can serve as a reference to discriminate different analytes within a single sensing platform. Furthermore, besides the intrinsic sensitivity and selectivity of Au NPs to melamine-like compounds, the “INH” gates of melamine/cysteine and melamine/Hg2+ in this logic system can be employed for sensitive and selective detections of cysteine and Hg2+, respectively.

Short-chain fatty acids cause an IL-8 response in cystic fibrosis airways via increased GPR41

RATIONALE: Anaerobic bacteria are present in large numbers in the airways of people with cystic fibrosis (PWCF). In the gut, anaerobes produce short-chain fatty acids (SCFAs) that modulate immune/inflammatory processes.

OBJECTIVES: To investigate the capacity of anaerobes to contribute to CF airway pathogenesis via SCFAs.

METHODS: Samples from 109 PWCF were processed using anaerobic microbiological culture with bacteria present identified by 16S RNA sequencing. SCFAs levels in anaerobe supernatants and bronchoalveolar lavage (BAL) were determined by gas chromatography. The mRNA and/or protein expression of SCFAs receptors, GPR41 and GPR43, in CF and non-CF bronchial brushings, and 16HBE14o- and CFBE41o- cells were evaluated using RT-PCR, western blot, laser scanning cytometry and confocal microscopy. SCFAs-induced IL-8 secretion was monitored by ELISA.

MEASUREMENTS AND MAIN RESULTS: Fifty seven of 109 (52.3%) PWCF were anaerobe-positive. Prevalence increased with age, from 33.3% to 57.7% in PWCF under (n=24) and over 6 years (n=85). All evaluated anaerobes produced millimolar concentrations of SCFAs, including acetic, propionic and butyric acid. SCFAs levels were higher in BAL samples from adults than children. GPR41 levels were elevated in; CFBE41o- versus 16HBE14o- cells; CF versus non-CF bronchial brushings; 16HBE14o- cells after treatment with CFTR inhibitor CFTR(inh)-172, CF BAL, or inducers of endoplasmic reticulum stress. SCFAs induced a dose-dependent and pertussis toxin-sensitive IL-8 response in bronchial epithelial cells with a higher production of IL-8 in CFBE41o- than 16HBE14o- cells.

CONCLUSIONS: This study illustrates that SCFAs contribute to excessive production of IL-8 in CF airways colonized with anaerobes via upregulated GPR41.

Spray-dried polysaccharide microparticles aimed at pulmonary delivery of antitubercular drugs

Dissertação de mestrado, Ciências Biomédicas, Departamento de Ciências Biomédicas e Medicina, Universidade do Algarve, 2015

Preservación de la anchoveta a bordo de las embarcaciones pesqueras, con fines industriales de reducción (nivel de laboratorio)

Describe el estudio de preservación de la anchoveta a nivel de laboratorio, a bordo de las embarcaciones pesqueras, empleando diferentes reactivos químicos (nitrito de sodio, INH-100, benzoato de sodio), en comparación con los métodos de refrigeración con hielo y con agua de mar refrigerada. Así mismo, también se indican las recomendaciones para el desarrollo de la etapa a nivel industrial que comprende: estudios para determinar los remanentes de los reactivos preservadores en la harina y los compuestos tóxicos que pudieron formarse durante el procesamiento; el rendimiento y calidad de los productos (harina y aceite) y los estudios de nutrición y toxicología.

Bovine follicle development is associated with divergent changes in activin-A, inhibin-A and follistatin and the relative abundance of different follistatin isoforms in follicular fluid

The aim was to determine whether follicle growth in cattle is accompanied by changes in levels of inhibin-A (inh-A), activin-A (act-A) and different Mr isofomus of follistatin (FS) in bovine follicular fluid (bFF), reflecting differential roles of these proteins during folliculogenesis. Follcles (n= 146) from 2-20 min diameter were dissected from ovaries of similar to 40 cattle. Immunoassays were used to measure total FS, act-A, inh-A, oestradiol (E) and progesterone (P) levels; immunoblotting was used to quanti, the relative abundance of different FS isoforms. Follicle growth from 2-6 mm was associated with a 6-fold increase in inh-A and 30-fold increase in act-A; FS remained uniformly high from 2-10 turn. From 6-2 min, inh-A remained high while act-A and FS fell 3-fold and 2-fold, respectively. Act-A/FS ratio increased 20-fold from 2-6 mm before falling slightly through to 20 mm. Act-A/inh-A ratio increased 6-fold from 2-6 nun before falling 2-fold from 6 to 17-20 mm. These findings imply a marked increase in relative activin 'tone' around the stage at which dominant follicle,;election occurs. When larger follicles (13-20 mm) were subdivided according to E/P ratio, those with high (> 5) E/P ratio had lower (2-fold; P < 0(.)001) levels of inh-A and act-A in comparison to follicles with low (< 5) E/P ratio, but there were no significant diffierences in FS, act-A/inh-A ratio or act-A/FS ratio. Thus follicle size, but not oestrogenic status, has a major influence on the intrafollicular balance between act-A and its opposing factors, inh-A and FS. Six FS isoforms were detected in bFF (apparent Mr: 65, 41, 37, 35, 33 and 31 kDa) averaging 6, 13, 24, 26 13 and 17% respectively of total FS. During growth from 2-20 mm the proportion of total FS represented by 605, 41 and 37 kDa isoforms increased similar to 2-fold while the proportion represented by the 33 and 31 kDa isoforms decreased by 3-fold and 1(.)6-fold, respectively. Treatment of bovine granulosa cells in vitro with FSH and IGF alone or in combination increased total FS secretion up to 12-fold but did not affect the relative abundance of the five different FS isoforms detected. While the functional significance of the intriguing shift in FS isoform abundance in bFF during follicle development remains to be established, we have shown that a marked increase in intrafollicular activin 'tone' accompanies bovine follicle growth from 3-6 min, corresponding to the stage at which the FSH-dependent follicle selection mechanism operates in this species.