Atrophic femoral nonunion successfully treated with teriparatide.

Teriparatide is a synthetic polypeptide hormone that contains the 1-34 aminoacid fragment of the recombinant human parathyroid hormone. It has been approved for the treatment of postmenopausal women with osteoporosis who are at high risk for sustaining a fragility fracture. It has been shown that teriparatide also accelerates fracture healing by improving the biomechanical properties of the fracture callus, increasing endochondral ossification and bone remodeling in animal models. This effect has been observed in several case reports. Fracture healing disorders negatively affect the patient's quality of life and result in high healthcare costs, as a second surgery is required to stabilize the fracture and stimulate bone biology. Future biotechnologies that accelerate fracture healing may be useful tools. We present a case report of delayed union of a femoral fracture treated with teriparatide. She was diagnosed with right distal metaphyseal femoral fracture on total knee arthroplasty. She underwent surgery at our center consisting of ORIF with lateral femoral locking plate in October 2011. Radiologic controls at 5 and 7 months did not show any signs of healing. After 2 months of treatment with teriparatide, the X-ray showed the presence of bone bridges and a decreased gap between fragments and a different aspect of neoformed bone. After 3 months of treatment, healing was complete. Our case report seems to confirm the possible effect of TPTD as bone induction through a more rapid healing of fractures. The TPTD could have a potentially important role in treating some forms of nonunion and delay in consolidation. Thus, one could hypothesize the possibility of a medical treatment with TPTD both as a preventive way and also as a support to the synthesis in high risk of nonunion fractures and complexed fractures in osteoporotic bone.

Resveratrol proniosomes as a convenient nanoingredient for functional food.

Proniosomes are free-flowing powders composed of water-soluble carriers blended with surfactants, which form niosomes upon hydration. In this work, proniosomal formulations containing the natural antioxidant resveratrol (RSV) were prepared and fully characterized. A pre-formulation study on RSV-loaded niosomes was carried out to determine the most promising ratio between the two surfactants, Tween 20 and Span 60, in terms of entrapment efficiency and antioxidant activity. The optimized formulae were subsequently adapted to be prepared as proniosomes by the slurry method, including lactose or maltodextrin as carriers. The impact of surfactants and carriers properties on size, entrapment efficiency and release kinetics of proniosomes were evaluated. In vitro release of RSV in simulated gastric and intestinal media was determined, as well as the vesicular stability. Moreover, the biocompatibility of the formulations was determined on intestinal cells in vitro. Overall, the developed proniosomes provide promising nanoingredient for functional food, improving resveratrol stability and bioavailability.

Immunosenescence and immunogenetics of human longevity.

At present, individuals can live up to 80-120 years, a time much longer than that of our ancestors, as a consequence of the improvements in life conditions and medical care. Thus, the human immune system has to cope with a lifelong and evolutionarily unpredicted exposure to a variety of antigens, which are at the basis of profound age-related changes globally indicated as immunosenescence, a multifaceted phenomenon that increases morbidity and mortality due to infections and age-related pathologies. The major changes occurring during immunosenescence are the result of the accumulation of cellular, molecular defects and involutive phenomena (such as thymic involution) occurring concomitantly to a hyperstimulation of both innate and adaptive immunity (accumulation of expanded clones of memory and effector T cells, shrinkage of the T cell receptor repertoire, progressive activation of macrophages), and resulting in a low-grade, chronic state of inflammation defined as inflammaging. It is unknown whether inflammaging, which represents a risk factor for most age-related pathologies, is a cause or rather an effect of the aging process. In this complex scenario, the role of genetic background likely represents a fundamental variable to attain successful aging and longevity. Accordingly, centenarians seem to be equipped with gene variants that allow them to optimize the balance between pro- and anti-inflammatory molecules, and thus to minimize the effects of the lifelong exposure to environmental insults and stressors. The remarkable features of the genetics of aging and longevity are reviewed, stressing the unexpected and unusual results obtained regarding such a postreproductive type of genetics.

GM-CSF production in human adenocarcinoma cell lines.

We describe the production of human granulocyte-macrophage colony-stimulating factor (hGM-CSF), by cell lines established from patients with different stages of breast, lung and colon adenocarcinoma. GM-CSF expression has been identified by immunocytochemistry determination, quantified on conditioned medium with specific ELISA procedure and evaluated by means of proliferation and differentiation of normal human monocytic and granulocytic progenitors. The growth of cell lines after incubation with exogenous GM-CSF and antibody-antiGM-CSF was not modified. To better understand the patho-physiologic role of hGM-CSF in vivo we also estimated its serum levels at diagnosis in 75 patients with breast lung and colon adenocarcinoma and in 69 healthy person. Only two patients showed detectable GM-CSF levels. The lack of growth modulation observed in vitro with exogenous GM-CSF and antibody anti-GM-CSF suggests a non autocrine secretion by adenocarcinoma cells. The serum investigation evidences that the leukocytosis observed in adenocarcinoma patients is unrelated to a GM-CSF constitutive tumor production in vivo.

Carotenoids in bird plumage: the complement of red pigments in the plumage of wild and captive bullfinch (Pyrrhula pyrrhula).

We have studied the carotenoid pigments in the red plumage of male bullfinch (Pyrrhula pyrrhula) immediately following capture and after the completion of the moult in captivity under dietary control. Astaxanthin, adonirubin, and alpha-doradexanthin, as well as papilioeritrinone and canthaxanthin (in lower amounts) are in every case the dominant carotenoids in the plumage pigment of wild individuals. alpha-Doradexanthin is responsible for the reddish-rose colour, which captive individuals adopt after a diet consisting mainly of lutein as disposable carotenoid. The red pigmentation biogenesis of captive bullfinch is compared with those of other red pigmented Carduelinae in which male individuals usually lose the red colour in captivity, namely Carpodacus roseus, Carpodacus rubricilloides, Uragus sibiricus, Carduelis cannabina, Carduelis flammea, Loxia curvirostra and Pinicola enucleator.

The chemical structure of the pigments in Ara macao plumage.

Parrots (Psittaciformes) harbor unusually bright, non-carotenoid, feather pigments. We successfully extracted and purified a sufficient quantity of pigment from the red plumage of the Scarlet Macaw (Ara macao) for a partial chemical analysis. The extracts were analyzed by HPLC coupled with UV-VIS and mass spectroscopy before and after total hydrogenation. We found at least four pigment components. We propose a linear polyenal structure comparable with the molecules tetradecahexenal, hexadecaheptenal, octadecaoctenal and eicosanonenal.

Synergism between gamma interferon and doxorubicin in a human MDR colon adenocarcinoma cell line.

Recent interest in cancer therapy derives from the ability of interferons to synergistically increase the activity of chemotherapeutic agents. To understand the biological basis of this synergism we evaluated the effects of human recombinant IFN-gamma on the expression of the mdr1 gene and on the cellular growth of a human colon adenocarcinoma cell line (LoVo) and its MDR subline (LoVo/Dx) after coincubation with doxorubicin. Treatment with IFN-gamma showed unchanged levels of MDR1-glycoprotein, no perturbation on cell cycle distribution and a significant reduction of colony formation in both lines (P < 0.05) starting from 100 U/ml. A synergistic effect was observed in the LoVo/Dx cell line when doxorubicin was added after exposure to 0.1-10 U/ml of IFN-gamma. Our data indicate that the effects of IFN-gamma, independent from action on cell proliferation and from modulation of p-glycoprotein expression, are a cause of the synergistic activity between this lymphokine and conventional chemotherapeutic agents such as doxorubicin.

Biological activity of parrodienes, a new class of polyunsaturated linear aldehydes similar to carotenoids.

A new chemical named parrodienes has been extracted from parrots' plumage. From the chemical point of view, parrodienes are polyunsaturated aldehydes similar to carotenoids. On the basis of this similarity we organized some biological experiments to evaluate the inhibition of lipoperoxidation of cell membranes induced by CCl4, protection against ultraviolet rays, anti-inflammatory activity and protection from an increase of ornithine-decarboxylase as marker of tumoral skin alteration. The results of these experiments showed that unsaturated dienes (parrodienes) play an important role on the inhibition and prevention of many biological processes that are at the basis of different pathological disorders.

Biologic characterization of pleural metastases from lung adenocarcinoma: description of the new DV90 cell line.

AIMS AND BACKGROUND: The characterization of pleural metastases from lung adenocarcinoma is often limited to single biologic features. METHODS: The present paper describes the cellular kinetic parameters, as well as immunocytochemical, ultrastructural and genetic characteristics of the new DV90 cell line, established from the pleural effusion of a stage IV lung adenocarcinoma. RESULTS: The cell line has a diploid DNA content, a doubling time of 24 h and 7% cloning efficiency, it is tumorigenic in nude mice. Ultrastructural investigation revealed the typical features of lung adenocarcinoma; the diagnosis was confirmed by its immunohistochemical reactivity with a panel of monoclonal antibodies specifically capable of identifying adenocarcinoma cells. Genetic analysis revealed a 46 X, -Y, +8, der (6)t(6?)(q27;?) karyotype and hyperexpression of the protein codified by genes Her2/Neu and p53. CONCLUSION: The importance of multidisciplinary biologic characterization in identifying the origin and biological behavior of pleural metastases deriving from lung adenocarcinoma is discussed.

[Our experience with biosynthetic human insulin]. / La nostra esperienza con insulina umana biosintetica.

Biosynthetic Human Insulin (BHI) was administered to 20 diabetics for six months. A clinical examination, glycaemia before and after eating, total glycosuria-ketonuria, and glycosylated haemoglobin tests were performed at fixed intervals, and the results were statistically analysed. It is concluded that human insulin produces no significant side effects. More important that the type of insulin treatment is effective outpatients surveillance of the diabetic, to ensure an adequate metabolic compensation.

[Correlation between the metabolic follow-up and diabetic retinopathy]. / Correlazione tra stato di controllo metabolico e retinopatia diabetica.

The state of the fundus oculi was investigated in 40 insulin-dependent and 20 non-insulin dependent diabetics. In both groups the state of the fundus oculi was correlated with total cholesterol, LDL, HDL, glycosylated haemoglobin, platelet adhesiveness, triglycerides and duration of the disease. In the case of insulin-dependent diabetics a correlation was also sought with daily doses of insulin. The fundus oculi was normal in 21/40 insulin-dependent and 14/20 non-insulin-dependent subjects. Multiple regression showed a positive correlation between insulin-HbA doses and duration of the disease and the fundus oculi. Simple liner regression showed a negative correlation between insulin doses and fundus, a tendentially positive correlation between platelet adhesion and fundus. A positive correlation between LDL-platelet adhesion and fundus was also shown in the non-insulin-dependent subjects. These results suggest that factors held to influence macroangiopathy also have an effect on macroangiopathy, that the check up of diabetics is important for the prevention of retinopathy, that insulin doses should be strictly limited to the necessary amount for the control of diabetes since excess is as harmful as insufficiency.

Synthetic biological response modifiers. Part II. Synthesis and immunomodulatory properties of some: N2-[omega-(pyrimidin-1-yl)carboxyalkyl]-L-arginines.

The synthesis and chemical and physical properties of some N2-[omega-(pyrimidin-1-yl)carboxyalkyl]-L-arginines are here described. Experiments of immunopharmacological evaluations are also described.

[A cytofluorimetric analysis of the DNA content and S phase in intraepithelial neoplasia of the cervix uteri]. / Analisi citofluorimetrica del contenuto in DNA e fase S nelle neoplasie intraepiteliali della cervice uterina.

The natural history of cervical intraepithelial neoplasia (CIN) evidenced in such pathologies a different biological behaviour due to the presence of similar morphological atypia with different potential development. To biologically characterize CIN lesions we valued by flow cytometry, the DNA content and the proliferative activity of 53 biopsies obtained by colposcopy. Aneuploid histograms were present in none of the histologically negative lesions, in 14% of the CIN I, in none CIN II, and in 40% of the CIN III. The mean value of S-phase was 2.7% in non dysplastic lesions, 5% in the CIN I, 2.5% in the CIN II and 6.1% in the CIN III. In this study the findings of aneuploid cells and the value of proliferative activities seems to be not correlated with the histological features. The clinical and instrumental follow up of the considered patients could establish the eventual prognostic significance of cytofluorimetric parameters in CIN lesions.

[Role of colposcopy in high-grade CIN (CIN II-III)]. / Il ruolo della colposcopia nelle CIN di grado elevato (CIN II-III).

We have considered the colpo-cytologic characteristic of 83 patients with CIN II-III histologic lesions over 900 colposcopic biopsies carried out at our Department from 1990-1992. In particular 38 cases were classified as CIN II of which 23 associated with HPV cytopathic feature, while 45 cases were, classified as CIN III, of which 13 associated with HPV c.f. 29% of CIN II were evident in women under 30 years of age; in this group the age decreased with the presence of HPV. 31% of CIN III were present in women under 35. A good correlation between cytologic and histological analysis on the same patient was observed particularly in CIN with the higher grade. Also a good correlation between colposcopy grading and CIN was observed. In CIN II, grade I images were present, while in CIN III, punctuation and white epithelium were the most common features. Our study shows also the impossibility of distinguishing between the images of simple viral infection and their related CIN morphologic patterns. Colposcopy represents a basic test for the definition of CIN, particularly for those with higher grade, and a complementary test for the definition of the topography of the lesions with the correct choice of the therapeutic treatment.

Synthesis of purine derivatives of potential immunomodulatory activity.

Moving from the interest as immunomodulatory agent of ST789 was studied the synthesis of series of N9alkylated hypoxanthine and adenine. The synthesis and the chemical physical properties of these derivatives are here described.

Determination of substances of abuse in biological fluids by GC/MS using chemical ionization with acetonitrile.

GC/ms is an important method to determine drugs in biological fluids. For each of the most commonly used drugs a methanolic solution (100 micrograms/ml) was prepared; 30 microliters of each solution was evaporated using nitrogen flow and than diluted in 100 microliters of a derivatizing mixture constituted by MSTFA and toluene (30/70). After a short reaction time at room temperature 1 ml of each solution was injected into a SE-52 column (15 m x 0.25 mm) using chemical ionization with acetonitrile. This method has a good signal/noise ratio and is very useful in screening techniques.

Immune parameters identify Italian centenarians with a longer five-year survival independent of their health and functional status.

Centenarians are rare and exceptional individuals characterized by a peculiar phenotype. They are the best example of healthy aging in humans as most of them have escaped or substantially delayed the onset of major age-related diseases. Within this scenario, the purpose of the present work was to understand if immune status is associated with survival and health status in centenarians. To this aim, 116 centenarians were concomitantly characterized for their immunological, health and functional status, and followed-up for five-year survival. On the basis of previous knowledge we focused on a core of fundamental and basic immune parameters (number of leukocytes, monocytes, total lymphocytes, CD3(+) T lymphocytes, CD4(+) helper T lymphocytes, CD8(+) cytotoxic T lymphocytes, CD19(+) B lymphocytes and plasma levels of IgM), and the most important findings can be summarized as follows: i. a hierarchical cluster analysis was able to define Cluster1 (88 centenarians) and Cluster2 (28 centenarians) characterized by low and high values of all these immune parameters, respectively; ii. centenarians of Cluster2 showed a statistically longer five-year survival and more favorable values of other important immune (naïve, activated/memory and effector/memory T cells) and metabolic (glycemia, insulin and HOMA-IR) parameters, in accord with previous observations that centenarians have a peculiar immune profile, a preserved insulin pathway and a lower incidence of type 2 diabetes; and iii. unexpectedly, parameters related to frailty, as well as functional and cognitive status, did not show any significant correlation with the immune clustering, despite being capable per se of predicting survival. In conclusion, high values of basic immunological parameters and important T cell subsets correlate with five-year survival in centenarians, independent of other phenotypic characteristics. This unexpected biological scenario is compatible with the general hypothesis that in centenarians a progressive disconnection and loss of biological coherence among the different functions of the body occur, where survival/mortality result from the failure of any of these domains which apparently follow an independent age-related trajectory.

Does the longevity of one or both parents influence the health status of their offspring?

According to the findings of some recent studies, the centenarians' offspring appear to represent a promising model for research on longevity and healthy aging. This study compares the health status and the functional status of three groups of subjects: 1. individuals with two long-lived parents (one of whom centenarian), 2. individuals with only one long-lived (centenarian) parent, and 3. individuals with no long-lived parents. The goal is to verify whether the centenarians' offspring display any advantage over the offspring of both non-long-lived parents and to evaluate whether the longevity of the non-centenarian parent provides a further advantage. A total of 374 subjects (mean age approximately 70 years) was examined. A threshold for longevity was established for non-centenarian parents through demographic data available for Italy (males surviving to at least 81 years of age and females to 87 years). The participants were assessed for their health and functional status by means of a standardized questionnaire and tests of physical performance. Data were analyzed using multivariate regression models adjusted for socio-demographic characteristics and risk factors for age-related pathologies. The results of the study show that centenarians' offspring have a better functional status, a reduced risk for several age-related pathologies and reduced drug consumption than the offspring of non-long-lived parents. In addition, the health status of centenarians' offspring does not appear to be influenced by the longevity of the second parent. It therefore seems possible to conclude that at ages around 70 years the genetic contribution to health status deriving from having one centenarian parent is not substantially improved if the other parent is also long-lived.

Metabolic syndrome in the offspring of centenarians: focus on prevalence, components, and adipokines.

With aging, an increased prevalence of a clustering of metabolic abnormalities has been observed. These abnormalities include obesity, dyslipidemia, hypertension, and insulin resistance and are collectively known as metabolic syndrome (MetS), a low-grade, systemic, inflammatory condition associated with an increased risk of cardiovascular disease, diabetes, and other adverse health outcomes. A number of studies have demonstrated that centenarians' offspring have a significant survival advantage and a lower risk of developing the most important age-related diseases. They therefore represent one of the best models with which to study the familiar component of human longevity. The aim of this study was to determine if the offspring of centenarians (n = 265 subjects) showed a different prevalence of MetS in comparison to the offspring of non-long-lived parents (controls, n = 101 subjects). In addition, we assessed whether centenarians' offspring showed particular features of MetS and a distinct regulation of circulating adipokines, cytokines, and metabolic mediators. Although the prevalence of MetS was quite similar both in the offspring of centenarians and the controls, MetS-affected centenarians' offspring seemed healthier, more functionally fit, and had lower resistin levels. MetS prevalence did not change in centenarians' offspring across resistin, IGF-1, and resistin/IGF-1 ratio tertiles. On the other hand, in controls, MetS prevalence strongly increased across resistin tertiles and in the third resistin/IGF-1 ratio tertile, indicating a dramatic increase in MetS prevalence when the ratio between these two factors is unbalanced, with high levels of resistin and low levels of IGF-1.

Nanocrystals as tool to improve piroxicam dissolution rate in novel orally disintegrating tablets.

In this paper, orally disintegrating tablets (ODT) were prepared using nanocrystal formulations in order to optimise dissolution properties of lipophilic, poorly soluble drug piroxicam (PRX). Different nanocrystal formulations were prepared using a high pressure homogenisation technique and poloxamer 188 as stabiliser. Characterisation of PRX nanocrystal ODT was carried out by infrared spectroscopy (FTIR), X-ray powder diffractometry (XRPD), differential scanning calorimetry and photon correlation spectroscopy. Dissolution study of PRX ODT was performed in distilled water (pH 5.5) and was compared to that of PRX coarse suspension ODT, PRX/poloxamer 188 physical mixture and bulk PRX samples. The XRPD and FTIR studies demonstrated that the homogenisation process led to a polymorphic transition from form I (bulk commercial PRX) to form III and monohydrate form of the nanocrystals. All ODT formulations prepared using PRX nanosuspensions showed a higher PRX dissolution rate compared with the ODT prepared with the coarse PRX. Since the solubility of the different PRX polymorphic forms increased only slightly from bulk PRX (form I) to monohydrate, form II and form III, we can conclude that the improvement in PRX dissolution rate is mainly caused by the increased surface-to-volume ratio due to the submicron dimension of the drug particles.