O fenômeno do extermínio de crianças e adolescentes no Rio de Janeiro: um estudo dos homicídios a partir dos anos 90

O presente estudo trata, a partir de uma abordagem crítica do debate no campo dos Direitos Humanos, da análise do extermínio de crianças e adolescentes no Rio de Janeiro, tendo em perspectiva que esses homicídios retratam a criminalização da pobreza como principal mecanismo de ação do Estado diante do agravamento da questão social e suas expressões. Inicialmente, a apresentação do arcabouço conceitual referencia as particularidades das violações dos direitos de crianças e adolescentes no contexto da formação sociohistórica brasileira, trazendo o debate na constituição do mito das classes perigosas. Em seguida procuramos compreender o contexto da crise capitalista contemporânea e a trajetória da consecução de direitos para crianças e adolescentes, circunscritos entre os avanços legais e o adensamento do processo de criminalização da infância e juventude pauperizada. Concluímos com a análise dos índices de homicídios da Região Metropolitana, a partir dos anos 90, abordando-os como resultado de um processo sistemático de extermínio de crianças e adolescentes, a culminância da violência cotidiana a qual estão submetidas, em contraposição a doutrina da proteção integral e da universalização de direitos.

Efeito da restrição alimentar materna durante a lactação sobre a função ovariana da prole fêmea na puberdade e vida adulta

α.O objetivo deste estudo foi avaliar se a restrição alimentar materna durante a lactação altera a função ovariana da prole na puberdade e na vida adulta. No dia do nascimento da ninhada, as ratas foram separadas aleatoriamente nos seguintes grupos: (C) controle = dieta com 23% de proteína; (PER) restrição protéica calórica = dieta com 8% de proteína. Após o desmame, as proles tiveram livre acesso a dieta com 23% proteína e apenas os animais na fase diestro foram sacrificados com uma dose letal de pentobarbital aos 40 e 90 dias de idade. O sangue foi retirado por punção cardíaca e o soro armazendo para posterior dosagem dos níveis hormonais por radioimunoensaio. O ovário direito foi excisado, pesado e armazenado a 80C para subseqüente avaliação do receptor de androgênio (AR), das isoformas dos receptores de estrogênio (ERα, ERβ1 e ERβ2), do receptor do hormônio folículo estimulante (FSHR), do receptor do hormonio luteinizante (LHR), das isoformas dos receptores de leptina (Ob-R, Ob-Ra and Ob-Rb), e da enzima aromatase pela técnica de RT-PCR. O ovário esquerdo foi incluido em parafina, seccionado em cortes de 5 μm de espessura e processado por análise histológica de rotina, para classificação dos foliculos ovarianos. Na puberdade, o grupo PER apresentou um aumento significativo (p<0,05) no número dos folículos pré-antrais e antrais, enquanto o número de folículos primordiais, folículos de Graaf e corpo lúteo foram reduzidos significativamente (p<0,05). As concentrações sericas de estradiol (pg/ml) foram significativamente aumentadas (p<0,05). Houve aumento significativo na expressão do RNAm dos FSHR (p<0.05) e LHR (p<0.05), e diminuição significativa na expressão do RNAm dos AR (p<0.05), ERα (p<0.05), ERβ1 (p<0.05) e ERβ2 (p<0.05). Na vida adulta, a restrição alimentar causou uma diminuição no número total de folículos ovarianos, entretanto esta redução só foi significativa no número dos folículos primordiais, primários e de Graaf (p<0.05). Houve redução significativa da expressão da enzima aromatase (p<0.01), do FSHR (p<0.05), LHR (p<0.05), Ob-R (p<0.05) e Ob-Rb (p<0.05). Podemos concluir que a restrição alimentar materna durante a lactação causa uma programação metabólica no ovário da prole, alterando a foliculogênese, expressão das diferentes isoformas dos receptores de estrogênio, leptina, androgênio e gonadotropinas e da enzima aromatase. Essa programação, possivelmente decorrente dos baixos níveis de leptina nos primeiros dias de vida, pode contribuir para uma senescência precoce e redução da fertilidade já descrita na literatura.

Age and length composition of Columbia Basin chinook, sockeye, and coho salmon at Bonneville Dam in 2001

In 2001, representative samples of adult Columbia Basin chinook (Oncorhynchus tshawytscha), sockeye (O. nerka), and coho salmon (O. kisutch) populations at Bonneville Dam were collected. Fish were trapped, anesthetized, sampled for scales and biological data, revived, and then released adult migrating salmonids. Scales were examined to estimate age composition; the results contributed to an ongoing database for age class structure of Columbia Basin salmon populations. Based on scale analysis of chinook salmon, four-year-old fish (from brood year [BY] 1997) comprised 88% of the spring chinook, 67% of the summer chinook, and 42% of the Bright fall chinook salmon population. Five-year-old fish (BY 1996) comprised 9% of the spring chinook, 14% of the summer chinook, and 9% of the fall chinook salmon population. The sockeye salmon population at Bonneville was predominantly four-year-old fish (81%), with 18% returning as five-year-olds in 2001. The coho salmon population was 96% three-year-old fish (Age 1.1). Length analysis of the 2001 returns indicated that chinook salmon with a stream-type life history are larger (mean length) than the chinook salmon with an ocean-type life history. Trends in mean length over the sampling period for returning 2001 chinook salmon were analyzed. Chinook salmon of age classes 0.2 and 1.3 show a significant increase in mean length over time. Age classes 0.1, 0.3, 0.4, 1.1, 1.2, and 1.4 show no significant change over time. A year class regression over the past 12 years of data was used to predict spring, summer, and Bright fall chinook salmon population sizes for 2002. Based on three-year-old returns, the relationship predicts four-year-old returns of 132,600 (± 46,300, 90% predictive interval [PI]) spring chinook and 44,200 (± 11,700, 90% PI) summer chinook salmon for the 2002 runs. Based on four-year-old returns, the relationship predicts five-year-old returns of 87,800 (± 54,500, 90% PI) spring, 33,500 (± 11,500, 90% PI) summer, and 77,100 (± 25,800, 90% PI) Bright fall chinook salmon for the 2002 runs. The 2002 run size predictions should be used with caution; some of these predictions are well beyond the range of previously observed data.

Age and length composition of Columbia Basin chinook, sockeye, and coho salmon at Bonneville Dam in 2000

In 2000, representative samples of adult Columbia Basin chinook (Oncorhynchus tshawytscha), sockeye (O. nerka), and coho salmon (O. kisutch), populations were collected at Bonneville Dam. Fish were trapped, anesthetized, sampled for scales and biological data, allowed to revive, and then released. Scales were examined to estimate age composition and the results contribute to an ongoing database for age class structure of Columbia Basin salmon populations. Based on scale analysis, four-year-old fish (from brood year (BY) 1996) were estimated to comprise 83% of the spring chinook, 31% of the summer chinook, and 32% of the upriver bright fall chinook salmon population. Five-year-old fish (BY 1995) were estimated to comprise 2% of the spring chinook, 26% of the summer chinook, and 40% of the fall chinook salmon population. Three-year-old fish (BY 1997) were estimated to comprise 14% of the spring chinook, 42% of the summer chinook, and 17% of the fall chinook salmon population. Two-year-olds accounted for approximately 11% of the fall chinook population. The sockeye salmon population sampled at Bonneville was predominantly four-year-old fish (95%), and the coho salmon population was 99.9% three-year-old fish (Age 1.1). Length analysis of the 2000 returns indicated that chinook salmon with a stream-type life history are larger (mean length) than the chinook salmon with an ocean-type life history. Trends in mean length over the sampling period were also analysis for returning 2000 chinook salmon. Fish of age classes 0.2, 1.1, 1.2, and 1.3 have a significant increase in mean length over time. Age classes 0.3 and 0.4 have no significant change over time and age 0.1 chinook salmon had a significant decrease in mean length over time. A year class regression over the past 11 years of data was used to predict spring and summer chinook salmon population sizes for 2001. Based on three-year-old returns, the relationship predicts four-year-old returns of 325,000 (± 111,600, 90% Predictive Interval [PI]) spring chinook and 27,800 (± 29,750, 90% PI) summer chinook salmon. Based on four-year-old returns, the relationship predicts five-year-old returns of 54,300 (± 40,600, 90% PI) spring chinook and 11,000 (± 3,250, 90% PI) summer chinook salmon. The 2001 run size predictions used in this report should be used with caution, these predictions are well beyond the range of previously observed data.

Diode laser pumped, KERR phase-shifting and switching of a diode laser signal in a small area optical fibre

In this paper authors report the first demonstration of a diode laser powered Kerr effect device, consisting of a single birefringent fiber, able to phase-shift and switch an optical signal generated by a second laser diode. They have obtained fast, stable phase-shifting of 90° in a single fiber, at a coupled pump power of only 20 mW. Using this phase shift to induce polarization switching with resultant gating, 25% modulation of the diode laser signal has been observed, with a detection limited-rise time of 10ns.

Microwave signal transmission over a directly-modulated radio-over-fiber link using cascaded semiconductor optical amplifiers

We demonstrate a record 150km transmission of microwave signals by a directly-modulated radio-over-fiber link with a bit-error-rate of less than 10-12. Cascaded semiconductor optical amplifiers are employed in this link to extend the transmission link length. © 2005 Optical Society of America.

ENSO and precipitation variability over Mexico during the last 90 years

Latin America has been shown to be susceptible to climatic anomalies during El Niño/Southern Oscillation (ENSO) events (eg, Aceituno 1988; Ropelewshi and Halpert 1987; Kiladis and Diaz 1989). While these studies have emphasized ENSO-related rainfall and temperature anomalies over Central and South America, less work has been done on the climatic effects of ENSO over the Mexican region. In this study we are investigating interannual and intraseasonal fluctuation in temperature and precipitation over the southwestern United States and Mexico since the turn of the century. We are particularly interested in the effects of ENSO on the interannual variability over this region. This report focuses on the association between ENSO and interannual variability of precipitation over Mexico.

Annual estimates of the unobserved incidental kill of pantropical spotted dolphin (Stenella attenuata attenuata) calves in the tuna purse-seine fishery of the eastern tropical Pacific

We estimated the total number of pantropical spotted dolphin (Stenella attenuata) mothers killed without their calves (“calf deficit”) in all tuna purse-seine sets from 1973– 90 and 1996–2000 in the eastern tropical Pacific. Estimates were based on a tally of the mothers killed as reported by color pattern and gender, several color-pattern-based frequency tables, and a weaning model. Over the time series, there was a decrease in the calf deficit from approximately 2800 for the western-southern stock and 5000 in the northeastern stock to about 60 missing calves per year. The mean deficit per set decreased from approximately 1.5 missing calves per set in the mid-1970s to 0.01 per set in the late-1990s. Over the time series examined, from 75% to 95% of the lactating females killed were killed without a calf. Under the assumption that these orphaned calves did not survive without their mothers, this calf deficit represents an approximately 14% increase in the reported kill of calves, which is relatively constant across the years examined. Because the calf deficit as we have defined it is based on the kill of mothers, the total number of missing calves that we estimate is potentially an underestimate of the actual number killed. Further research on the mechanism by which separation of mother and calf occurs is required to obtain better estimates of the unobserved kill of dolphin calves in this fishery.

缺失nifZ的棕色固氮菌突变株DJ194的钼铁蛋白研究—纯化、结晶及金属簇的组成与分布

从棕色固氮菌DJ194菌株得到的固氮酶粗提液经DEAE－52、Sephacryl S－200及Q－Sepharose等柱厌氧层析，分离纯化得到nifZ基因缺失的固氮酶MoFe蛋白（ΔnifZ Av1）制备物。通过天然电泳和SDS变性电泳发现早期纯化所得的ΔnifZ Av1制备物中残存相当比例的三个主要污染蛋白：属于热激蛋白60家族（Hsp 60 family）成员的分子伴侣GroEL、糖酵解过程的一个多功能酶——6-磷酸葡萄糖异构酶（6-Phosphate Glucose Isomerase，PGI）及棕色固氮菌细菌铁蛋白（Bacterioferritin，Bfr）。初步鉴定表明，它们分别为由约55kD亚基组成的14聚合体，62kD亚基组成的10聚体和20kD亚基组成的24聚体。首次发现PGI有如此高的聚合体。虽然GroEL和PGI在天然电泳中的迁移率小于ΔnifZ Av1蛋白，但它们的亚基在SDS变性电泳中与ΔnifZ Av1亚基具有相似的迁移率，互相重叠，从而使变性电泳比天然电泳显出更高的ΔnifZ Av1纯度;而细菌铁蛋白虽然不会在变性电泳中污染ΔnifZ Av1，但往往会在ΔnifZ Av1制备物的结晶中优先或较多地结晶出来,从而给它的晶体生长和解析研究带来干扰（Zhao等，2004）。 通过Sephacryl S－200柱洗脱峰收集精度的调整及Q－Sepharose柱的NaCl浓度梯度洗脱，得到了纯度大于90%的ΔnifZ Av1制备物。它的厌氧天然电泳及其免疫印渍（Western blotting），以及SDS-变性凝胶电泳显出，ΔnifZ Av1的电泳迁移率、分子量和亚基组成等均与野生种钼铁蛋白（OP Av1）相似，表明nifZ基因缺失并未改变ΔnifZ Av1的α2β2四聚体构成。ΔnifZ Av1的Mo含量、EPR信号（g≈4.3, 3.65和2.01）和520-660 nm附近的圆二色摩尔消光系数（Δε）也都与OP Av1较相似，从而表明ΔnifZ Av1含有与OP Av1数量相当的具有3/2自旋态的还原FeMoco。然而，ΔnifZ Av1的Fe含量和对底物（C2H2、H+和N2）的还原活性都较低, 分别约为OP Av1的74%和46-50%;而反映P-cluster状况的450nm附近的Δε也明显低于OP Av1。此外，与OP Av1相同的ΔnifZ Av1在g≈2.01的EPR信号却与推测含由双[4Fe-4S]簇组成的P-cluster前体的His-tagged ΔnifZ Av1（Hu等，2004）的信号明显不同。这就表明，ΔnifZ Av1与OP Av1的差别不在于FeMoco的结构、含量和氧还状态，也不在于P-cluster的结构和氧还状态，而仅在于ΔnifZ Av1中P-cluster数目的减少（约一半）。据此推测出与国外提出的His-tagged ΔnifZ Av1模型不同的ΔnifZ Av1(DJ194)的如下结构模型。一个αβ亚基对含有一个FeMoco和一个P-cluster，而另一个αβ亚基对只含FeMoco，其P-cluster区域则是空的。由于nifZ基因的缺失只造成了钼铁蛋白中两个P-cluster中的一个不能组装，因此推测P-cluster的组装可能不是受单一基因产物的影响。根据Lee等（1998）对nifZ产物（NifZ）和nifW产物（NifW）可以形成[NifWx-NifZy]多聚体，并可能是通过同一途径来影响固氮酶的合成的研究，提出NifZ的如下的可能作用机理。[NifWx-NifZy]多聚体影响与P-cluster合成相关的金属簇(如[4Fe-4S])的进入和P-cluster的最终合成，而其中的一个αβ对上的P-cluster的形成可能较多的受到NifZ的影响;nifZ的某些突变或缺失虽然不影响[NifWx-NifZy]多聚体的形成，但对于更依赖于NifZ的那个αβ对上的P-cluster的合成可能会产生不同的影响。 对这一高纯度的ΔnifZ Av1制备物结晶的研究表明，使用Tris缓冲系统的25%PEG 6K/MgCl2晶体培养液可以在较短的晶体培养时间内得到较大的晶体;在一定条件下，pH为7.5和8.3的培养液对出晶数和晶体大小的影响不明显;PEG 6K的浓度与MgCl2的浓度对出晶大小的影响有一定的相关性，即较低的PEG浓度在较低的MgCl2浓度下和较高的PEG浓度搭配较高的MgCl2浓度较易长出较大的晶体。虽然ΔnifZ Av1制备物的纯度达到90%以上，并且在染铁实验中表明其基本不含细菌铁蛋白，但我们在对得到的晶体进行电泳鉴定中仍然发现了一种与以前报导的砖红色晶体不同的深棕红色的细菌铁蛋白晶体，之所以颜色不同可能和所含的铁元素的氧化状态有关。不过在所鉴定的5支结晶管中只在一个管内发现了与固氮酶晶体同时存在的这种细菌铁蛋白晶体，说明通过蛋白纯度的提高减少细菌铁蛋白的含量以及晶体培养条件的优化，细菌铁蛋白晶体形成的几率就可大大降低。

青蒿素生物合成相关基因对烟草遗传转化的研究

　　青蒿素是存在于中药青蒿(Artemisia annua L.)中的一种含有过氧桥的倍半萜内酯化合物，是中国科学家研发出的当今最有潜力的抗疟药剂，较传统抗疟药很少或无毒副作用，因此青蒿素的生产备受人们关注。目前，青蒿素的生产主要以植物提取为主，但由于青蒿植株中青蒿素的含量很低（约占干重的0.01%～0.8%），从而导致青蒿素价格昂贵，使许多贫困地区的疟疾患者无法得到医治，故提高青蒿植株中青蒿素的含量或扩大青蒿素的来源，降低生产青蒿素的成本具有重要的意义。　　 　　本论文基于扩大青蒿素的来源和提高青蒿植株中青蒿素含量的目的，开展了以下两方面的工作： 一、紫穗槐二烯在烟草中组合生物合成的研究 　　紫穗槐二烯合酶（amorpha-4,11-diene synthase，ADS）是青蒿素生物合成的关键酶之一，为了能在烟草中合成青蒿素的前体，本研究将青蒿的紫穗槐二烯合酶基因置于CaMV 35S启动子控制下，通过根癌农杆菌介导转入烟草（Nicotiana tobacum L.），并获得了转ADS基因烟草植株。经PCR及Southern杂交分析表明，ADS基因已经整合到转基因烟草基因组中；RT-PCR及对转基因烟草中ADS酶活性和产物中紫穗槐二烯和植物甾醇的测定分析，进一步证明整合的ADS基因在转录、翻译水平上均已经表达。上述结果表明，利用基因工程将青蒿素生物合成途径的关键酶基因导入植物，转基因植物中能够合成青蒿素的前体，这一研究结果为利用转基因植物生产青蒿素或其前体奠定了基础。 二、青蒿鲨烯合酶双链干涉基因对烟草的遗传转化研究 　　鲨烯合酶（squalene synthase, SQS）是甾醇类生物合成分支途径的关键酶之一，利用RNA干扰技术(RNA interference，RNAi)抑制目标基因表达的技术已日趋成熟。本文根据植物中hpRNA(hairpin RNA)的原理，在与烟草SQS同源性高达80％的青蒿ASQS序列的5/端保守区选择622 bp作为构建RNAi的序列，借助中间克隆载体，经过三次亚克隆，最后形成含ASQS－RNAi表达盒的双元表达载体pART27－ASQS，并转入农杆菌EHA105。采用农杆菌介导的烟草叶盘转化法，共获得了12棵转基因植株。转基因植株经过PCR和PCR-Southern blotting 检测，证实外源ASQS基因已经导入烟草中，并已经成功整合到烟草基因组中；通过RT-PCR分析说明，转基因烟草中SQS基因的表达已被成功抑制，部分转基因植株中内源SQS的干扰效果高达90％以上。对SQS的直接产物鲨烯和最终产物植物甾醇的检测显示，转基因烟草的植物甾醇和鲨烯的含量明显低于对照。本实验的结果为下一步将此RNA干扰载体导入青蒿，抑制青蒿中ASQS基因的表达，从而提高青蒿素的含量提供了可能。

植物分子伴侣热休克蛋白90（Hsp90）的分子作用机理及抗逆基因工程研究

热激蛋白90是广泛存在于各类细菌和真核生物中的一类高度保守的分子伴侣，它对维持细胞生命是绝对必需的。对Hsp90的相关认知主要来源于对动物和酵母细胞的研究，植物Hsp90的研究甚少。由于植物的特殊性，因此对植物Hsp90的研究是对Hsp90未知功能的有力补充。拟南芥中有7个Hsp90蛋白，其中AtHsp90-1、AtHsp90-2、AtHsp90-3和AtHsp90-4定位在细胞质中，AtHsp90-5、AtHsp90-6和AtHsp90-7分别定位在叶绿体、线粒体和内质网中。本文对拟南芥中的AtHsp90-1、AtHsp90-2、AtHsp90-5、AtHsp90-6和AtHsp90-7五个基因进行了克隆，并分别利用酵母互补、双杂交和拟南芥过表达体系几个层面进行了功能分析。 我们利用酵母穿梭载体p416GPD构建了五个AtHsp90基因的酵母表达载体，将其转入Hsp90基因点突变和条件型缺失的酵母菌株iG170D和R0005中。酵母功能互补实验表明细胞质定位的AtHsp90-1和AtHsp90-2可以在各种胁迫条件下互补酵母Hsp90的功能，而定位于细胞器中的AtHsp90-5、 AtHsp90-6和AtHsp90- 7则在任何条件下都不能互补酵母Hsp90的功能。我们还对转基因酵母进行了液体培养的动态观测和细胞膜完整性检测，其结果和固体培养的结果一致。这说明细胞质Hsp90的功能具有一定的保守性，细胞器Hsp90的功能有其特殊性。 热激蛋白90在执行其生物功能时，需要和大量的辅助因子相互作用，因此我们利用酵母双杂交体系检测了AtHsp90-1、AtHsp90-2、AtHsp90-5、AtHsp90-6和AtHsp90-7五个Hsp90蛋白和Hsp70、p23、Cyp40、NOS等几个辅助因子之间的相互作用情况。双杂交结果显示AtHsp90-1和AtHsp90-2几乎不和所选的这几个辅助因子相互作用，AtHsp90-5可以和所有的辅助因子相互作用、AtHsp90-6可以和除Hsp70以外的辅助因子相互作用，AtHsp90-7也可以和所有的辅助因子相互作用但和Hsp70及Hsp70t-2和互作较其他辅助因子弱一些。可以看出胞质Hsp90和细胞器Hsp90在和辅助因子相互作用时有一定的差异。 为了进一步了解拟南芥个Hsp90基因在抗非生物逆境中的作用，我们又将AtHsp90-2、AtHsp90-5、AtHsp90-7基因插入植物表达载体pBI121，用农杆菌介导的浸蕾法将这三个基因转入拟南芥并在其中过量表达，并研究了这些基因的过表达植株的种子和幼苗对多种模拟非生物逆境的响应。结果显示，转基因种子和幼苗对ABA、盐(NaCl)、干旱(甘露醇)、高温、氧化、高钙等非生物逆境都表现出了敏感，转细胞器Hsp90的种子和幼苗比转细胞质Hsp90的更为敏感。但在高浓度钙离子胁迫下，幼苗表现情况与盐、旱和氧化等非生物逆境处理下的情况正好相反，转细胞器Hsp90的幼苗比转细胞质Hsp90的长得健壮。这些结果表明Hsp90参与了植物抵抗非生物逆境的反应，其作用可能是通过ABA和Ca2+途径实现的，然而体内Hsp90的动态平衡可能才是植物抵抗非生物逆境的关键。