Genetic dissection of dietary restriction in mice supports the metabolic efficiency model of life extension

Dietary restriction (DR) has been used for decades to retard aging in rodents, but its mechanism of action remains an enigma. A principal roadblock has been that DR affects many different processes, making it difficult to distinguish cause and effect. To address this problem, we applied a quantitative genetics approach utilizing the ILSXISS series of mouse recombinant inbred strains. Across 42 strains, mean female lifespan ranged from 380 to 1070 days on DR (fed 60% of ad libitum [AL]) and from 490 to 1020 days on an AL diet. Longevity under DR and AL is under genetic control, showing 34% and 36% heritability, respectively. There was no correlation between lifespans on DR and AL; thus different genes modulate longevity under the two regimens. DR lifespans are significantly correlated with female fertility after return to an AL diet after various periods of DR (R = 0.44, P = 0.006). We assessed fuel efficiency (FE, ability to maintain growth and body weight independent of absolute food intake) using a multivariate approach and found it to be correlated with longevity and female fertility, suggesting possible causality. We found several quantitative trait loci responsible for these traits, mapping to chromosomes 7, 9, and 15. We present a metabolic model in which the anti-aging effects of DR are consistent with the ability to efficiently utilize dietary resources.     

T-cell function is critical for murine cholesterol gallstone formation

BACKGROUND & AIMS: The formation of cholesterol gallstones is a complex process involving contributions from genes and environmental factors. Although gallbladder inflammation is believed to be common during cholelithogenesis, the role of immunologic factors is unknown. METHODS: The role of adaptive immunity in cholesterol cholelithogenesis was analyzed utilizing immunocompetent Helicobacter spp.-infected and -uninfected BALB/c and congenic immunodeficient Rag2(-/-) (Rag) mice. Lymphocyte transfer studies were performed to determine which cellular subset was responsible for cholesterol gallstone formation. Also, gallbladder inflammation was quantified to determine the nature of the inflammatory response associated with cholelilithogenesis. RESULTS: When fed a lithogenic diet for 8 weeks, wild-type mice developed significantly more cholesterol gallstones (27%-80% prevalence) than Rag mice ( approximately 5%, P < .05). Helicobacter spp.-infected BALB/cJ mice displayed statistically significant increases in cholesterol gallstone prevalence compared with uninfected mice (81% vs. 39%; P < .05). Transfer of splenocytes or T lymphocytes to Rag2(-/-) mice increased stone prevalence markedly (26% and 40% respectively; P < .05), whereas transfer of B cells was not appreciably cholelithogenic (13%). The adaptive immune response increased the expression of gallbladder Muc genes and accumulation of mucin gel. In addition, T cells and cholesterol monohydrate crystals induced proinflammatory gene expression in the gallbladder, which likely contributes to gallbladder dysfunction. CONCLUSIONS: These studies indicate that T cells are critical in murine cholesterol cholelithogenesis. Furthermore, cholesterol monohydrate crystals induce expression of proinflammatory cytokines in a T-cell-dependent fashion. Acquired immunity and inflammation are likely to be crucial factors in cholesterol gallstone pathogenesis, rather then merely the result of cholelithogenesis.     

Controlled release of huperzine A from biodegradable microspheres: In vitro and in vivo studies

This paper describes the effect on Sun Protection Factor (SPF) of the combination of inorganic and organic filters in sunscreen products as determined by an in vitro method. O/W emulsions containing inorganic filters, such as titanium dioxide and zinc oxide, combined with 18 EU-authorized UV-B organic filters were tested. SPF measurements were carried out using a spectrophotometer equipped with an integrating sphere.

This study observed a synergic effect when titanium dioxide was combined with either anisotriazine or octyldimethylPABA. The combination of zinc oxide with 11 UV-B organic filters also exhibited a similar synergy; however, the measured SPF was systematically lower than the protection factor achieved with titanium dioxide.     

Proteomic analysis identifies candidate proteins associated with distant recurrences in breast cancer after adjuvant chemotherapy

Breast cancer is a heterogeneous disease and it is of importance to select patients with regard to different prognosis and treatment sensitivity to individualize treatment regimes. In this study we successfully adapted a protein extraction protocol from mRNA extracted tumor samples enabling two-dimensional gel electrophoresis (2-DE) analysis of samples previously analyzed by cDNA microarray. The aim was to find candidate proteins that distinguish breast cancer patients with or without recurrences after adjuvant CMF (cyclophosphamide, methotrexate and 5-FU) treatment within four years to follow-up. We identified several proteins distinguishing the recurrence group from the non-recurrence group, especially in the ER and PgR positive subgroup (n=7). The induced proteins were involved in translation/folding, iron ion binding, and protease inhibition, whereas proteins involved in signaling, ubiquitination, and splicing were decreased in expression. These results show that it is possible to use 2-DE to separate high abundant proteins in breast cancer tissue and to find discriminating proteins to identify patients with different prognosis after adjuvant CMF treatment.     

The TIP39–PTH2 receptor system: Unique peptidergic cell groups in the brainstem and their interactions with central regulatory mechanisms

Tuberoinfundibular peptide of 39 residues (TIP39) is the recently purified endogenous ligand of the previously orphan G-protein coupled parathyroid hormone 2 receptor (PTH2R). The TIP39–PTH2R system is a unique neuropeptide–receptor system whose localization and functions in the central nervous system are different from any other neuropeptides. TIP39 is expressed in two brain regions, the subparafascicular area in the posterior thalamus, and the medial paralemniscal nucleus in the lateral pons. Subparafascicular TIP39 neurons seem to divide into a medial and a lateral cell population in the periventricular gray of the thalamus, and in the posterior intralaminar complex of the thalamus, respectively. Periventricular thalamic TIP39 neurons project mostly to limbic brain regions, the posterior intralaminar thalamic TIP39 neurons to neuroendocrine brain areas, and the medial paralemniscal TIP39 neurons to auditory and other brainstem regions, and the spinal cord. The widely distributed axon terminals of TIP39 neurons have a similar distribution as the PTH2R-containing neurons, and their fibers, providing the anatomical basis of a neuromodulatory action of TIP39. Initial functional studies implicated the TIP39–PTH2R system in nociceptive information processing in the spinal cord, in the regulation of different hypophysiotropic neurons in the hypothalamus, and in the modulation of affective behaviors. Recently developed novel experimental tools including mice with targeted mutations of the TIP39–PTH2R system and specific antagonists of the PTH2R will further facilitate the identification of the specific roles of TIP39 and the PTH2R.     

A new intranasal influenza vaccine based on a novel polycationic lipid-ceramide carbamoyl-spermine (CCS). II. Studies in mice and ferrets and mechanism of adjuvanticity

We recently showed that lipid assemblies comprised of a novel polycationic sphingolipid (ceramide carbamoyl-spermine, CCS) are an effective adjuvant/carrier when complexed with cholesterol (CCS/C) for influenza and other vaccines administered parenterally and intranasally (i.n.) in mice. Here we expand these studies to ferrets, an established model of influenza infection. We also address the question of why the CCS/C-based liposomal vaccine (also known as VaxiSome™) in mice is superior to vaccines based on liposomes of other lipid compositions (neutral, anionic or cationic). Ferrets immunized i.n. with CCS/C-influenza vaccine produced significantly higher hemagglutination inhibition (HI) antibody titers compared to ferrets immunized intramuscularly with the unadjuvanted influenza vaccine, indicating that the CCS/C-based vaccine is very immunogenic. Furthermore, the i.n. adjuvanted vaccine was shown to significantly reduce the severity of influenza virus infection in ferrets following homologous viral challenge as determined by weight loss, temperature rise and viral titer. No adverse reactions were observed. Pharmacokinetic and biodistribution studies following i.n. administration in mice of CCS/C-based vaccine showed that both the lipids and antigens are retained in the nose and lung for at least 24 h, and it appears that this retention correlates with the superior immunogenicity elicited by the adjuvanted vaccine formulation. The CCS lipid also increases production of cytokines (mainly IFN gamma, IL-2 and IL-12) and co-stimulatory molecules’ expression, which might further explain the robust adjuvantation of this liposome-based vaccine.     

小儿恶性肿瘤细胞的DNA含量及其临床意义

目的：探讨小儿恶性肿瘤ＤＮＡ含量及其与临床预后的关系。方法：用流式细胞仪对６２例小儿恶性肿瘤细胞的ＤＮＡ含量进行测定，并分析其中３５例的细胞增殖状况及其与临床分期的关系。结果：ＤＮＡ组方图呈二倍体的肿瘤１４例，异倍体的肿瘤４８例（检出率７７．４％）；Ⅲ～Ⅳ期肿瘤的异倍体检出率高达９７．４％；ＤＮＡ指数（ＤＩ）和倍性与肿瘤的分型无关。结论：测定结果应与临床、病理资料结合，对预后作出准确判断     

金黄地鼠封闭群SPF化后的微卫星遗传学检测

目的检测和评估金黄地鼠封闭群SPF化后的遗传学变化,为SPF金黄地鼠遗传质量的控制提供技术资料。方法应用小鼠和大鼠的微卫星标记筛选适于金黄地鼠遗传检测的微卫星标记,并结合微卫星荧光标记-半自动基因分型技术,对成都生物制品研究所的SPF级金黄地鼠及其来源的普通级金黄地鼠进行遗传检测,计算其群体遗传学参数。结果对18个小鼠和6个大鼠微卫星标记进行了筛选,分别有2个小鼠和2个大鼠微卫星标记在金黄地鼠种群中具PCR扩增多态性。4个检测的微卫星位点在普通级金黄地鼠和SPF金黄地鼠种群分别发现25和20个等位基因,两群体的期望杂合度分别为0.4979和0.5048,其群体遗传多样性无显著差异;群体间的不同微卫星位点FST范围从0.0095到0.0367,平均为0.0315,表明两群体间的遗传分化很弱,其遗传多样性主要存在于群体内;Nei(1972)遗传距离和Nei(1978)无偏遗传距离分别为0.0678和0.0570,表明了2群体之间很高的遗传相似度和非常近的亲缘关系;Hardy-Weinberg平衡检验表明普通级和SPF金黄地鼠分别有2个和3个位点偏离遗传平衡,且偏离位点均表现为杂合子缺陷。结论该SPF金黄地鼠基本保持了其来源普通级黄地鼠的遗传多样性,两群体间遗传分化程度和遗传差异很小,但应进一步加强其封闭群的繁育控制,保持其遗传稳定性。     

Prognostic value of DNA flow cytometry in paragangliomas of the carotid body

A carotid body tumor is a paraganglioma of the carotid bifurcation. Histologic appearance does not correlate with the malignant potential of the lesion, and thus a reliable prognostic marker for these tumors is lacking. To determine whether flow cytometric analysis of paraffin block specimens by DNA index and synthetic phase fraction (SPF) would be of prognostic value, a retrospective chart review of 14 patients with carotid body tumors was performed. Three of 14 tumors were aneuploid and were the only tumors with SPF greater than 15%. One of 3 patients (33%) with an aneuploid tumor (SPF = 22%) developed a local recurrence; no patient with a diploid tumor developed a recurrence. Two of 3 (67%) patients with aneuploid tumors (SPF = 18%) but only 1 of 11 (9%) with a diploid tumor were symptomatic preoperatively (P = 0.03). DNA index and SPF may help select a subgroup of patients with more aggressive tumors who are at increased risk for recurrence and therefore require closer follow-up.     

SPF级和普通级新西兰兔主要脏器系数的比较

目的测定3月龄SPF级和普通级新西兰兔的主要脏器系数，并进行对比研究。方法选用3月龄SPF级和普通级新西兰兔各30只（♂15只，♀15只），用电子天平分别测定心、肝、脾、肺、肾、肾上腺、胃、睾丸、附睾和子宫的脏器系数。结果（1）SPF级新西兰兔的主要脏器系数与普通级新西兰兔的相比，除了肾上腺的脏器系数二者之间差异不显著之外（P〉0．05），前者其余主要脏器的脏器系数均显著低于后者（P〈0．01）；（2）SPF级新西兰雄兔除了肝、脾、肾的脏器系数显著高于雌兔之外（P〈0．01），雄兔其余脏器的脏器系数与雌兔相比差异均不存在显著性（P〉0．05）。结论不同环境条件和饲养管理对SPF级和普通级新西兰兔主要脏器系数有一定影响。