Thymus transplantation, a critical factor for correction of autoimmune disease in aging MRL/+mice.

MRL/MP-+/+ (MRL/+) mice develop pancreatitis and sialoadenitis after they reach 7 months of age. Conventional bone marrow transplantation has been found to be ineffective in the treatment of these forms of apparent autoimmune disease. Old MRL/+ mice show a dramatic thymic involution with age. Hematolymphoid reconstitution is incomplete when fetal liver cells (as a source of hemopoietic stem cells) plus fetal bone (FB; which is used to recruit stromal cells) are transplanted from immunologically normal C57BL/6 donor mice to MRL/+ female recipients. Embryonic thymus from allogeneic C57BL/6 donors was therefore engrafted along with either bone marrow or fetal hematopoietic cells (FHCs) plus fragments of adult or fetal bone. More than seventy percent of old MRL/+ mice (> 7 months) that had been given a fetal thymus (FT) transplant plus either bone marrow or FHCs and also bone fragments survived more than 100 days after treatment. The mice that received FHCs, FB, plus FT from allogeneic donors developed normal T cell and B cell functions. Serum amylase levels decreased in these mice whereas they increased in the mice that received FHCs and FB but not FT. The pancreatitis and sialoadenitis already present at the time of transplantations were fully corrected according to histological analysis by transplants of allogeneic FHCs, FB and FT in the MRL/+ mice. These findings are taken as an experimental indication that perhaps stem cell transplants along with FT grafts might represent a useful strategy for treatment of autoimmune diseases in aged humans.

Prolactin increases CD4/CD8 cell ratio in thymus-grafted congenitally athymic nude mice.

One distinctive effect on T-cell development was analyzed by selectively increasing serum prolactin (PRL) concentration in thymus-grafted congenitally athymic nude mice and by neutralizing PRL in suspension cultures of thymus from 1-day-old neonatal mice. Flow cytometric analysis of single-positive CD4+ and CD8+ cells derived from inguinal lymph nodes revealed a CD4/CD8 cell ratio of 2.2 +/- 0.18 (mean +/- SEM) in thymus-grafted nude mice that is similar to the ratio for immune-competent BALB/c mice (2.0 +/- 0.06). Addition of the pituitary to thymus-grafted nude mice significantly elevated serum PRL (P < 0.005) and increased the CD4/CD8 cell ratio (2.8 +/- 0.12; P < 0.005), demonstrating preferential stimulation of CD4+ cell development. T cells in nude mice receiving sham (submandibular salivary gland) or pituitary grafts alone were below detectable levels. Suspension cultures of neonatal thymus treated with anti-mouse PRL antiserum resulted in 20% and 30% decreases in double-positive CD4+8+ thymocytes and thymocyte viability, respectively. A 10-fold increase in double-negative CD4-8- thymocytes expressing the interleukin 2 receptor alpha chain, CD25, was also observed concurrently. Our findings illustrate an important way in which PRL may participate in two interrelated mechanisms: the regulation of peripheral single-positive cells and the maintenance of thymocyte viability during the double-positive stage of intrathymic differentiation.

Inhibition of acute in vivo human immunodeficiency virus infection by human interleukin 10 treatment of SCID mice implanted with human fetal thymus and liver.

To improve the usefulness of in vivo mode for the investigation of the pathophysiology of human immunodeficiency virus (HIV) infection, we modified the construction of SCID mice implanted with human fetal thymus and liver (thy/liv-SCID-hu mice) so that the peripheral blood of the mice contained significant numbers of human monocytes and T cells. After inoculation with HIV-1(59), a primary patient isolate capable of infecting monocytes and T cells, the modified thy/liv-SCID-hu mice developed disseminated HIV infection that was associated with plasma viremia. The development of plasma viremia and HIV infection in thy/liv-SCID-hu mice inoculated with HIV-1(59) was inhibited by acute treatment with human interleukin (IL) 10 but not with human IL-12. The human peripheral blood mononuclear cells in these modified thy/liv-SCID-hu mice were responsive to in vivo treatment with exogenous cytokines. Human interferon gamma expression in the circulating human peripheral blood mononuclear cells was induced by treatment with IL-12 and inhibited by treatment with IL-10. Thus, these modified thy/liv-SCID-hu mice should prove to be a valuable in vivo model for examining the role of immunomodulatory therapy in modifying HIV infection. Furthermore, our demonstration of the vivo inhibitory effect of IL-10 on acute HIV infection suggests that further studies may be warranted to evaluate whether there is a role for IL-10 therapy in preventing HIV infection in individuals soon after exposure to HIV such as for children born to HIV-infected mothers.

Expression of the HPV16E7 oncoprotein by thymic epithelium is accompanied by disrupted T cell maturation and a failure of the thymus to involute with age

Transgenic mice expressing the E7 protein of HPV16 from the keratin 14 promoter demonstrate increasing thymic hypertrophy with age. This hypertrophy is associated with increased absolute numbers of all thymocyte types, and with increased cortical and medullary cellularity. In the thymic medulla, increased compartmentalization of the major thymic stromal cell types and expansion of thymic epithelial cell population is observed. Neither an increased rate of immature thymocyte division nor a decreased rate of immature thymocyte death was able to account for the observed hypertrophy. Thymocytes with reduced levels of expression of CD4 and/or CD8 were more abundant in transgenic (tg) mice and became increasingly more so with age. These thymic SP and DP populations with reduced levels of CD4 and/or CD8 markers had a lower rate of apoptosis in the tg than in the non-tg mice. The rate of export of mature thymocytes to peripheral lymphoid organs was less in tg animals relative to the pool of available mature cells, particularly for the increasingly abundant CD4lo population. We therefore suggest that mature thymocytes that would normally die in the thymus gradually accumulated in E7 transgenic animals, perhaps as a consequence of exposure to a hypertrophied E7-expressing thymic epithelium or to factors secreted by this expanded thymic stromal cell population. The K14E7 transgenic mouse thus provides a unique model to study effects of the thymic epithelial cell compartment on thymus development and involution.

Aberrant cervical thymus mimicking a cervical mass

A case of a 9-year-old female with suprasternal extension of the thymus mimicking thyroid gland enlargement is described. Ultrasonography successfully established the diagnosis. Aberrant cervical thymic tissue is an infrequently reported cause of paediatric neck masses. It is important to be aware of this entity to prevent anxiety and inappropriate investigation and/or intervention.

Control of proliferation in the rat thymus

Quiescent rat thymocytes were stimulated to divide by a variety of agents. One such mitogen was the neurotransmitter acetylcholine which exhibited a biphasic action. Interaction with low affinity nicotinic receptors was linked with an obligatory requirement for magnesium ions whereas combination with high affinity muscarinic receptors induced mitosis only if calcium ions were present in the medium. Binding of acetylcholine to its muscarinic receptor enhanced calcium influx and increased intracellular calcium levels causing calmodulin activation, a necessary prelude to DNA synthesis and mitosis. Nicotinic receptor activation may be associated with a magnesium influx and stimulation of cells in a calmodulin-independent fashion. Parathyroid hormone and its analogues exhibited only a monophasic mitogenic action. This response was linked to calcium influx, a rise in cytosolic calcium and calmodulin activation. Parathyroid hormone did not stimulate adenylate cyclase in thymocytes and decreased cellular cyclic AMP concentrations. Picomolar amounts of interleukin-2 (IL-2) also stimulated division in thymocytes derived from 3-month old rats by binding to high affinity receptors. The response in thymocytes from newborn and foetal animals was greater reflecting the larger proportion of cells bearing receptors at this age. The mitogenic effect of IL-2 was abolished by a monoclonal antibody directed against the IL-2 receptor. Injections of IL-2 itself or the administration of IL-2 secreting activated syngeneic spleen cells also stimulated proliferation of both thymus and bone marrow cells in vivo. Likewise immunisation with pertussis toxin, which enhances endogenous IL2 production, also increased mitosis in these tissues. Calcium influx, increased cytosolic Ca2+ levels and calmodulin activation are associated features of the mitogenic action of IL-2. Interleukin-1 was also found to be mitogenic in thymic lymphocyte cultures. The responses to this mitogen and to parathyroid hormone and acetylcholine were not inhibited by the anti-IL2 receptor antibody suggesting that the thymic lymphocyte bears discrete receptors for these agents. Subtle interactions of hormones, neurotransmitters and interleukins may thus contribute to the turnover and control of lymphoid cells in the thymus and perhaps bone-marrow.

Cognitive performance and the thymus among HIV-infected subjects receiving HAART

Objective: To evaluate the impact of alcohol use, which is widespread in human immunodeficiency virus (HIV)+ individuals, on highly active antiretroviral therapy (HAART)-associated immune and cognitive improvements and the relationship between those two responses. Methods: In a case-control longitudinal study, thymic volume, cognition, and immune responses were evaluated at baseline and after 6 months therapy in HIV+ and HIV- controls. Cognitive performance was evaluated using the HIV Dementia Score (HDS) and the California Verbal Learning Test (CVLT). Results: Prior to HAART, thymic volume varied considerably from 2.7 to 29.3 cm3 (11 ± 7.2 cm3). Thymic volume at baseline showed a significantly inverse correlation with the patient’s number of years of drinking (r2 = 0.207; p < 0.01), as well as HDS and the CVLT scores in both HIV-infected (r2 = 0.37, p = 0.03) and noninfected (r2 = 0.8, p = 0.01). HIV-infected individuals with a small thymic volume scored in the demented range, as compared with those with a larger thymus (7 ± 2.7 vs. 12 ± 2.3, p = 0.005). After HAART, light/moderate drinkers exhibited thymus size twice that of heavy drinkers (14.8 ± 10.4 vs. 6.9 ± 3.3 cm3). Conclusions: HAART-associated increases of thymus volume appear to be negatively affected by alcohol consumption and significantly related to their cognitive status. This result could have important clinical implications.

The impact of gamma irradiation on the cytotoxic properties and phenolic composition of Thymus vulgaris L. and Menta x piperita L.

Irradiation has been increasingly recognized as an effective decontamination technique, also ensuring the chemical and organoleptic quality of medicinal and aromatic plants 1 . The use of medicinal plants in the prevention and or treatment of several diseases has revealed satisfactory results as anti-inflammatory, antimutagenic, anti-cancer and antioxidant agents 2 . The aim of the present study was to evaluate the effects of gamma irradiation on the cytotoxic properties and phenolic composition of Thymus vulgaris L. and Menta x piperita L. (methanolic extracts). Phenolic compounds were analyzed by HPLC-DAD-ESI MS, while the cytotoxicity of the samples was assessed in MCF-7 (breast adenocarcinoma), NCI-H460 (non-small cell lung cancer), HeLa (cervical carcinoma), HepG2 (hepatocellular carcinoma) cell lines, as also in non-tumor cells (PLP2). Thirteen and fourteen phenolic compounds were detected in T. vulgaris and M. piperita, respectively, but none of them was affected by the irradiation up to a dose of 10 kGy. However, despite there were no changes in the cytotoxic properties of irradiated peppermint samples in tumor cell lines, the thyme samples irradiated with 10 kGy showed higher cytotoxicity in comparison with the samples submitted to other doses (2 and 5 kGy). This highlights that 10 kGy can be a suitable dose to ensure the sanitary treatment, without modifying the bioactive composition and properties of these aromatic plants.

Phenolic compounds and antioxidant capacity of three thymus species plants

Thymus plants comprise distinct species with claimed health properties [1], commonly associated to their essential oils and phenolic compounds. Albeit that, the phenolic composition and the biological activities of many Thymus species remain unclear. This work aimed to elucidate the phenolic composition and antioxidant properties of aqueous extracts from Thymus herba barona, Thymus caespetitus and Thymus fragrantissimus. The aqueous extracts of the three Thymus species were evaluated for their total phenolic compounds by an adaptation of the Folin-Ciocalteu method [2], and individual phenolic compounds were identified by high performance liquid chromatography associated with electrospray mass spectrometry (HPLC-DAD-ESI-MSn) in the negative mode. The antioxidant activity of each extract was carried out by DPPH● scavenging assay and ferric reducing antioxidant power assays [3]. Total phenolic compounds in the three extracts ranged from 236±27 (T. caespetitus) to 273±17 μg GAE/mg (T. fragrantissimus). Similarly to other Thymus species [1,4], these extracts were rich in caffeic acid derivatives (characteristic UV spectra maxima at 290 and 328 nm) and mainly composed of rosmarinic acid (MW 360). Other caffeic acid derivatives included salvianolic acid K (MW 556) and 3′-O-(8″-Z-caffeoyl)rosmarinic acid (MW 538). High amounts of the flavone luteolin-O-glucuronide ([M-H]− at m/z 461→285) were found in T. caespetitus while the others species contained moderate amounts of this compound. T. herba barona, T. caespetitus and T. fragrantissimus extracts showed high DPPH radical scavenge ability (EC50 values 11.6±0.9, 13.8±0.6 and 10.9±1.2 μg/mL respectively), as well as high reducing power (EC50 values of 35.1±4.5, 39.3±2.7 and 32.4±4.3 μg/mL, respectively), that were comparable to those of reference compounds. This work is an important contribution for the phytochemical characterization and the antioxidant capacity of these three Thymus species.

Phenolic compounds and antioxidant capacity of three thymus species plants

Thymus plants comprise distinct species with claimed health properties [1], commonly associated to their essential oils and phenolic compounds. Albeit that, the phenolic composition and the biological activities of many Thymus species remain unclear. This work aimed to elucidate the phenolic composition and antioxidant properties of aqueous extracts from Thymus herba barona, Thymus caespetitus and Thymus fragrantissimus. The aqueous extracts of the three Thymus species were evaluated for their total phenolic compounds by an adaptation of the Folin-Ciocalteu method [2], and individual phenolic compounds were identified by high performance liquid chromatography associated with electrospray mass spectrometry (HPLC-DAD-ESI-MSn) in the negative mode. The antioxidant activity of each extract was carried out by DPPH● scavenging assay and ferric reducing antioxidant power assays [3]. Total phenolic compounds in the three extracts ranged from 236±27 (T. caespetitus) to 273±17 μg GAE/mg (T. fragrantissimus). Similarly to other Thymus species [1,4], these extracts were rich in caffeic acid derivatives (characteristic UV spectra maxima at 290 and 328 nm) and mainly composed of rosmarinic acid (MW 360). Other caffeic acid derivatives included salvianolic acid K (MW 556) and 3′-O-(8″-Z-caffeoyl)rosmarinic acid (MW 538). High amounts of the flavone luteolin-O-glucuronide ([M-H]− at m/z 461→285) were found in T. caespetitus while the others species contained moderate amounts of this compound. T. herba barona, T. caespetitus and T. fragrantissimus extracts showed high DPPH radical scavenge ability (EC50 values 11.6±0.9, 13.8±0.6 and 10.9±1.2 μg/mL respectively), as well as high reducing power (EC50 values of 35.1±4.5, 39.3±2.7 and 32.4±4.3 μg/mL, respectively), that were comparable to those of reference compounds. This work is an important contribution for the phytochemical characterization and the antioxidant capacity of these three Thymus species.

Thymus vulgaris L. essential oil and its main component thymol: anthelmintic effects against Haemonchus contortus from sheep.

2016

Analysis Of The Dna Fourier Transform-infrared Microspectroscopic Signature Using An All-reflecting Objective.

The Fourier transform-infrared (FT-IR) signature of dry samples of DNA and DNA-polypeptide complexes, as studied by IR microspectroscopy using a diamond attenuated total reflection (ATR) objective, has revealed important discriminatory characteristics relative to the PO2(-) vibrational stretchings. However, DNA IR marks that provide information on the sample's richness in hydrogen bonds have not been resolved in the spectral profiles obtained with this objective. Here we investigated the performance of an all reflecting objective (ARO) for analysis of the FT-IR signal of hydrogen bonds in DNA samples differing in base richness types (salmon testis vs calf thymus). The results obtained using the ARO indicate prominent band peaks at the spectral region representative of the vibration of nitrogenous base hydrogen bonds and of NH and NH2 groups. The band areas at this spectral region differ in agreement with the DNA base richness type when using the ARO. A peak assigned to adenine was more evident in the AT-rich salmon DNA using either the ARO or the ATR objective. It is concluded that, for the discrimination of DNA IR hydrogen bond vibrations associated with varying base type proportions, the use of an ARO is recommended.

Endocarditis By Kocuria Rosea In An Immunocompetent Child.

Kocuria rosea belongs to genus Kocuria (Micrococcaceae family, suborder Micrococcineae, order Actinomycetales) that includes about 11 species of bacteria. Usually, Kocuria sp are commensal organisms that colonize oropharynx, skin and mucous membrane; Kocuria sp infections have been described in the last decade commonly affecting immunocompromised patients, using intravenous catheter or peritoneal dialysis. These patients had mainly bacteremia/recurrent sepsis. We hereby describe the case of a 10-year-old girl, immunocompetent, who had endocarditis/sepsis by K. rosea which was identified in five different blood cultures by Vitek 2 ID-GPC card (BioMérieux, France). Negative HIV serology, blood count within normal range of leukocytes/neutrophils and lymphocytes, normal fractions of the complement, normal level of immunoglobulins for the age; lymphocyte immunophenotyping was also within the expected values. Thymus image was normal at chest MRI. No catheters were required. Identification of K. rosea was essential to this case, allowing the differentiation of coagulase-negative staphylococci and use of an effective antibiotic treatment. Careful laboratory analysis of Gram-positive blood-born infections may reveal more cases of Kocuria sp infections in immunocompetent patients, which may collaborate for a better understanding, prevention and early treatment of these infections in pediatrics.

Vascularização arterial dos lobos torácicos do timo em fetos de suínos da Linhagem C40

As origens e distribuições das artérias que vascularizaram os lobos torácicos do timo foram estudadas em fetos de 30 suínos da linhagem C40, sendo 12 machos e 18 fêmeas. Os exemplares tiveram o sistema arterial preenchido com solução aquosa a 50% de Neoprene Látex corado e, em seguida, foram submetidos à fixação em solução aquosa a 10% de formaldeído. Os lobos torácicos do timo foram vascularizados por ramos diretos das artérias torácica interna direita (63,33%) e esquerda (53,33%), subclávia esquerda (3,33%), vertebral esquerda (3,33%), cervical superficial direita (3,33%) e esquerda (3,33%), carótida comum esquerda (3,33%), coronária direita (3,33%) e pelos troncos braquiocefálico (33,33%) e costocervical (3,33%). Observaram-se ainda os ramos indiretos das artérias torácica interna direita (70%) e esquerda (76,67%), subclávia esquerda (23,33%), cervical superficial esquerda (3,33%) e do arco aórtico (6,67%).