Gastric ghrelin, GOAT, leptin, and leptinR expression as well as peripheral serotonin are dysregulated in humans with obesity.

BACKGROUND: Gastrointestinal hormone release and the regulation of appetite and body weight are thought to be dysbalanced in obesity. However, human data investigating the expression of gastrointestinal hormones in the obese are rare. We studied the expression of ghrelin, leptin, and the serotonergic system in stomach tissue and serum of obese and non-obese individuals. METHODS: Gastric tissue and serum were collected from 29 adult obese (BMI 48.7 ± 10.6 kg/m(2) ; mean ± SD) who underwent laparoscopic sleeve gastrectomy. Gastric biopsies, surgery specimen or serum was obtained from 35 adult non-obese humans (BMI 22.7 ± 1.9 kg/m(2) ). Ghrelin, ghrelin O-acyl transferase (GOAT), leptin, leptin receptor, and tryptophan hydroxylase 1 (TPH1) mRNA expression were measured by qRT-PCR. Serotonin (5HT) and leptin protein concentration were quantified in tissue extracts and serum; GOAT and ghrelin-positive cells were immunohistologically quantified in tissue. Additionally, 21 blood immune markers were analyzed. KEY RESULTS: In gastric tissue, GOAT-positive cells were reduced (p < 0.01), but ghrelin-positive cells and mRNA were increased (both p < 0.05) in obese compared with non-obese individuals. Gastric leptin (p < 0.001) and leptin receptor (p < 0.001) mRNA expression, as well as leptin concentrations in serum (p < 0.001), were increased in obese compared with non-obese individuals. Serum 5HT was reduced (p < 0.05), while tissue 5HT and TPH1 mRNA were reduced only by trend. Interleukin 1 receptor a (IL1Ra), IL-8, IL-12, and monocyte chemoattractant protein 1 (IL1Ra) were increased and IL1Ra correlated negatively with serum leptin. CONCLUSIONS & INFERENCES: Our data indicate that obesity causes a dysregulation of gastrointestinal hormones at the tissue level and serum, including a negative correlation with an increased marker of subclinical inflammation.

Reference intervals for selected hematological and biochemical variables in Hucul horses.

BACKGROUND: Hucul horses are the unique, genetically distinct breed of Carpathian Mountains. Even though they are recognized as primitive breed, many morphological differences between them and other primitive horses have been reported. Neither hematological nor blood biochemical studies in this breed have been conducted so far. OBJECTIVES: The aim of this study was to establish the reference intervals for basic hematological and selected biochemical variables and to compare them with other breeds. MATERIAL AND METHODS: Blood samples were collected from 168 Hucul horses and the analyses were performed using routine methods. Mainly nonparametric method was used to establish reference intervals. RESULTS: The following reference intervals have been established (rounded to two significant digits): RBC: 7.0-13×1012/l; HGB: 106.1-195.8 g/l; HCT: 0.3-0.6 l/l; MCV: 35-50 fl; MCH 11.9-17.1 pg; MCHC: 31.9-34.8 g/dl; WBC: 7.5-22×109/l, bands: 0-0.5×109/l; segmented neutrophils: 3.3-10×109/l; eosinophils: 0-1.1×109/l; basophils: 0-0.3×109/l; lymphocytes: 1.9-12×109/l; monocytes: 0-0.2×109/l; PLT 95-350×109/l; MPV 5.2-7.0; ALP: 98-425 U/l; AST: 220-470 U/l; GGT: 9.1-31 U/l; total bilirubin: 6.5-29 µmol/l; CPK: 120-640 U/l; triglycerides: 0.1-0.9 mmol/l; urea: 3.8-11 mmol/l; creatinine: 44 -140 µmol/l; serum amyloid A: 130-5200 µg/l. CONCLUSIONS: Hematological and biochemical variables in Hucul horses were closer to hot-blooded then to cold-blooded and primitive horses or wild equidae. The reference intervals presented in this study pose clinically useful tool for evaluation of blood check-up in Hucul horses.

Serum amyloid A level as a potential indicator of the status of endurance horses.

REASONS FOR PERFORMING STUDY: Changes in serum levels of acute phase proteins (APPs) reflect the acute phase reaction, a rapid and nonspecific response to any tissue damage. Serum amyloid A (SAA) is the main APP in horses, which increases in various diseases, surgical injuries and after long distance endurance rides; however, this nonspecific parameter has not been investigated as an indicator of subclinical disorders, which may result in elimination during endurance competitions. OBJECTIVES: To evaluate the serum concentration of SAA as a potential indicator for the status of horses prepared for long distance endurance rides (120 and 160 km). MATERIALS AND METHODS: Twenty Arabian horses were tested and 12 were eliminated during the ride and 8 completed the distances. Routine haematological and biochemical tests and measurement of serum concentrations of SAA were carried out before and after the competition. Results were compared using the Mann Whitney U test. RESULTS: Before the competition all haematological and biochemical parameters varied within reference ranges with no differences between the eliminated horses and the ones that successfully finished the competition. After the rides creatine phosphokinase activity and neutrophil: lymphocyte ratio reflecting exercise-induced leukogram changes increased (P < 0.05) in both groups. Before the competition, the concentration of SAA remained within reference ranges; however, it was significantly (P < 0.05) lower in horses that successfully finished the competition than in eliminated ones (411.7 ± 144 ng/ml vs. 5809.5 ± 2242.7 ng/ml). After the ride SAA levels increased (P < 0.05) and were similar in both groups (13,833.8 ± 1354.3 ng/ml and 12,831.2 ± 1317.6 ng/ml). CONCLUSIONS: Serum SAA level was the only laboratory parameter that indicated most (66.6%) of the eliminated horses before entering the competition. None of the horses with SAA level higher than 1000 ng/ml completed the distance. Thus, it may be postulated that serum SAA concentration may indicate a poor status of a horse, resulting in elimination during a competition.

The effect of the 162 km endurance ride on equine peripheral blood neutrophil and lymphocyte functions.

Strenuous exercise is recognized as a stress, which may induce functional immunodeficiency and increase individual susceptibility to infection. It has been shown in equine athletes, that alterations in leukocyte functions occur after moderate and submaximal exertion, however, no data deal with the effect of extreme physical exertion. In this study, we evaluated leukocyte functions (neutrophil oxidative burst and lymphocyte proliferation activity in response to mitogens) in horses following the CEI 3* 162 km endurance ride. Exercise-induced stress was manifested as neutrophilic leukocytosis and lymphopaenia resulting in a significant increase in neutrophil:lymphocyte ratio. The alterations in neutrophil function were expressed as a lower percentage of the cells undergoing oxidative burst. The spontaneous lymphocyte proliferation was very high, however, the cells failed to respond to mitogens. Although a decrease in lymphocyte proliferation in response to mitogens has been reported previously, the pattern determined in our study was unique. It may suggest that during the extreme physical exercise immune cells receive an excessive stimulation from yet undetermined factor(s), which renders them unresponsive to extraneous mitogens. The differences between alterations in leukocyte activities induced by extreme exertion may reflect the exercise type and duration as well as the training status of the horses.

Immunobiology of bovine mammary gland: apoptosis of somatic cells in milk during naturally occurring mastitis.

Mastitis remains a major cause of economic losses in dairy herds. It is believed, that the enhancement of natural defense mechanisms in mammary gland may be helpful in the treatment of bovine mastitis. Our study was designed to assess the apoptosis of leukocytes isolated from bovine milk during subclinical staphylococcal mastitis. Milk samples were collected from cows naturally infected with one pathogen--Staphylococcus aureus and from animals with mastitis caused by several pathogens, including S. aureus. It has been determined that the rate of apoptosis was lower in mastitic milk, as compared with control samples, although varied significantly between groups. High percentage of apoptotic cells was detected in milk with high counts of pathogenic bacteria. In all groups the rate of apoptosis was dependent on the bacterial load, although various correlations were identified. Thus, it is postulated, that the rate of apoptosis of somatic cells in mastitic milk is related to the etiology of infection and is determined by the bacterial load.

Granulomatous inflammation during Heligmosomoides polygyrus primary infections in FVB mice.

Host responses to primary infections with Heligmosomoides polygyrus were studied in fast responding FVB mice (H-2(q)). Pathological changes in the intestinal mucosa, mesenteric lymph nodes and spleen were examined. Features of the fast response were typical: low effectiveness of infection and limiting of parasite survival and egg production, with worm expulsion occurring about 60 days post-infection. The intestinal inflammatory response involved infiltration by different cells into the intestinal mucosa and granulomata formation. As is typical for intestinal nematode infection enteropathy, decreased villus:crypt ratio and hyperplasia of goblet and Paneth cells were also present. Reactions of the intestinal mucosa, mesenteric lymph nodes and spleen increased over time post-infection and after worm expulsion. Enteropathy may help worm expulsion by creating an unfavourable environment for H. polygyrus. The implications of these findings and the potential role of intestinal intraepithelial lymphocytes in the pathogenesis of generated lesions are discussed.

Pathogenesis of mousepox in H-2(d) mice: evidence for MHC class I-restricted CD8(+) and MHC class II-restricted CD4(+) CTL antiviral activity in the lymph nodes, spleen and skin, but not in the conjunctivae.

Genetically sensitive mice (i.e. H-2(d) haplotype) infected with a natural mouse pathogen named ectromelia virus (EV) can develop a mousepox. Virus replicates well in the skin, next in the draining lymph nodes (DLNs) and then in the spleen and liver, where it may induce extensive necrosis with strong inflammatory reaction. It is well known from the studies defined on some other viruses that a correlation, functional link and powerful help exist between MHC class I-restricted CD8(+) and MHC class II-restricted CD4(+) virus-specific cytotoxic T lymphocytes (CTLs). However, in the case of mousepox the role of CD4(+) CTLs is still controversial and some reports support the notion that induction of EV-specific CD4(+) CTLs is nonessential for the generation of virus-specific immune response. Consequently, this study was designed to evaluate EV-specific CD8(+) and CD4(+) CTL activity in the DLNs, spleen, skin and conjunctivae of BALB/c (H-2(d)) mice at 7 and 14 days p.i. with Moscow strain of EV. By using bulk cytotoxicity assay and immunosurgery of effector T cells with mAb specific for CD4(+) and/or CD8(+) T cells our data show that EV-specific CD8(+) CTLs predominated in DLNs and spleen at 7 days (67 and 66% of total CTLs, respectively) and 14 days p.i. (63 and 69% of total CTLs, respectively). In contrast, we found that EV clearance from the cutaneous lesions during mousepox is CD4(+) CTL-dependent at 7 days p.i. (59% of total CTLs), whereas at 14 days p.i. CD8(+) CTLs predominated in the epidermis, accounting for 72% of the total EV-specific CTLs. Our studies showed that the population of EV-specific CTLs is heterogeneous and contains cells of both phenotypes: CD8(+) and CD4(+). However, these effector cells did not express a similar tendency in cytotoxic activity in the DLNs, spleen and skin in comparison to the conjunctivae where EV-specific CD8(+) and CD4(+) CTLs were not detected at 7 days p.i. and at peak of mousepox conjunctivitis (14 days p.i.). Our results are discussed in terms of the value of EV to study antiviral CTL responses in the genetically susceptible host.

Homing studies on distribution of ectromelia (mousepox) virus-specific T cells adoptively transferred into syngeneic H-2d mice: paradigm of lymphocyte migration.

Mousepox (infectious ectromelia) may be used as a model for studies on the cellular immune response and pathogenesis of generalized viral infections. Ectromelia virus (EV) initially replicates in the footpad (f.p.) skin at the site of infection, next in draining lymph nodes, and then in the spleen and liver where the virus may induce extensive necrotic process with inflammatory reaction. We show in this study that after recipient BALB/c mice (H-2d) f.p. infection with EV prior to the adoptive transfer of syngeneic donor EV-specific cytotoxic T lymphocytes interferon-gamma-positive (IFN-gamma-+), interleukin-2-positive (IL-2+), and IL-4+ of both phenotypes, CD8+ approximately 70%, and CD4+ approximately 30%) preferentially migrated to the inguinal and auxiliary lymph nodes, spleen, liver, and skin at the site of infection (f.p.). Many particles of EV with the morphology characteristic for orthopoxviruses and virus-specific immunofluorescence within the cells of inguinal and auxiliary lymph nodes, liver, spleen, and skin have been observed using high-resolution transmission electron microscopy and fluorescence antibody technique, respectively. Results presented in this article support the concept that immune T cells adoptively transferred into infected recipient mice are able not only to specific migration in the host and homing in the sites of virus replication, but also to develop immunoprotection in the transferred animals.

How human immunodeficiency viruses and herpesviruses affect apoptosis.

Eighty to hundred percent of patients positive for human immunodefficiency viruses 1 or 2 (HIV) may develop opportunistic viral infections. According to the National Institute of Health data, only in the USA the HIV patients are positive also for human cytomegalovirus (HCMV) in 25-40%, varicella-zoster virus (VZV) in 10%, herpes simplex viruses 1 and 2 (HSV-1 and HSV-2), and Kaposi's sarcoma-associated herpesvirus (human herpesvirus 8, KSHV, HHV-8) in 20%. HIV and herpesviruses express numerous different proteins that are able to influence interactions between the host and virus. One of the most interesting regulatory phenomenon is apoptosis which could play a significant role during both specific and non-specific antiviral response and latency. Apoptosis is an ordered cascade of precisely regulated enzymatic reactions which may be modulated or even controlled by viruses. Dramatic changes which occur during infection and which are exerted by HIV and certain herpesviruses on the mechanism of apoptosis may contribute to the pathogenesis of acquired immunodeficiency syndrome (AIDS).

[Regulation of the immune response to intestinal nematode infections in mice]. / Regulacja odpowiedzi immunologicznej na inwazje nicieni jelitowych u myszy.

Control of parasitic infections is dependent on mechanisms that limit invasion, reproduction or survival of the parasite, including elevated serum IgE, eosinophilia and intestinal mast cell hyperplasia. Studies with mice infected with Heligmosomoides polygyrus, Trichuris muris, Nippostrongylus brasiliensis and Trichinella spiralis have provided considerable information about immune mechanisms correlated with resistance and susceptibility. Activation and cytokine secretion of distinct Th cell subset leads to the generation of effective or ineffective responses resulting in clearance of the parasite load or maintenance of chronic infection. The induction of differential responses remains to be determined but is likely to be influenced at a number of levels including the host genetic background, involvement of accessory cells, activation of co-stimulatory molecules on antigen presenting cells. The regulation of responses to intestinal nematode infections is discussed.

Controlling orthopoxvirus infections--200 years after Jenner's revolutionary immunization.

An 11-year global WHO campaign for eradication of smallpox finished in October 1977 as the result of Edward Jenner's primary success in 1796, who for the first time applied human vaccination against variola virus (VARV). The 200th anniversary of this happening is a good occasion to summarize the current status of the knowledge about the role of B and T lymphocytes in the control of orthopoxvirus infections. This short review concentrates on general characteristics of orthopoxviruses and the immune response to infection, mainly by vaccinia virus (VV) and ectromelia virus (EV).

The inflammatory and immune response to mousepox (infectious ectromelia) virus.

The ectromelia virus (EV) has been recognized as the etiological agent of a relatively common infection in laboratory mouse colonies around the world, i.e., Europe (including Poland), USA and Asia. Due to widespread use of mice in biomedical research, it is important to study the biology of strains characteristic for a given country. This is particularly significant for the diagnosis, prevention and control ectromelia. In severe epizootics, approximately 90% morbidity is observed within colonies and mortality rate exceeding 70% is observed within 4 to 20 days from the appearance of clinical symptoms. The resistance to lethal infection is mouse strain-dependent. Several inbred strains of mice, including C57BL/6 and AKR are resistant to the lethal effects of EV infection, while others, such as A and BALB/c are susceptible. Recent studies indicate that (1) T lymphocytes, NK cells and interferon (IFN)-dependent host defenses must operate for the expression of resistance, (2) virus-specific T-cell precursors appear earlier in regional lymph nodes of resistant than susceptible mice, and (3) resistance mechanisms are expressed during early stages of infection. Over the past several years, (1) induction of anti-EV cytotoxic CD8+ T lymphocytes (CTL) responses in vivo in the absence of CD4+ (T helper) cells, (2) importance of some cytokines e.g., IFN-gamma in EV clearance at all stages of infection, and (3) induction of nitric oxide (NO) synthase, which is necessary for a substantial antiviral activity of IFN-gamma, have been demonstrated.(ABSTRACT TRUNCATED AT 250 WORDS)

Lymphocyte subpopulations in peripheral blood of pregnant sows.

Theophylline-sensitive, theophylline-resistant and theophylline-dependent populations of peripheral blood lymphocytes in pregnant sows were defined. Theophylline-sensitive lymphocytes were cautiously considered as helper and theophylline-resistant lymphocytes seemed to represent the population of suppressor cells. During the first two months of pregnancy the number of helpers increased, while decrease of suppressor cells during the second and third month of gestation was observed. Increase of population of helper T-cells was accompanied by increase of absolute number of B-lymphocytes. Rapid decrease of B-cells after farrowing accompanied the decline of suppressor population. The changes in percentage and in absolute number of these populations of immune cells do not indicate the general immunosuppression during pregnancy in sows.

Relationship between the count of milk somatic cells and the peripheral blood lymphocyte subpopulations in sows.

The aim of the study was to examine subpopulations of peripheral blood lymphocytes of mastitic and normal sows. Since number of cells in the milk reflects the intensity of response to mastitis process, relationship between level of milk somatic cells and peripheral blood lymphocytes was determined. No differences were found between mastitic and healthy sows in the percentage and absolute number of blood neutrophils and lymphocytes, T- and B-lymphocytes as well as in subpopulations of T-lymphocytes. It was found that milk somatic cell count positively correlates with the percentage but not with number of neutrophils and lymphoblasts and negatively correlates with the number of blood total T-lymphocytes and helper T-cells.

[Dynamics of homocytotropic antibody production in pigs]. / Próba okreslenia dynamiki wytwarzania przeciwcial homocytotropowych u swin.

Homocytotropic antibody (HTA) production in pigs was stimulated by sensitization with alum precipitated equine serum or with equine serum in Freund's complete adjuvant. Two doses of appropriate antigen were given in 7 days interval simultaneously with heat inactivated suspension of Bordetella pertussis organisms. HTA were detected in 6 of 10 pigs sensitized with alum precipitated serum and in 4 of 6 pigs sensitized with serum in Freund's complete adjuvant. In pigs sensitized with alum precipitated serum HTA appeared at day 6 after the first antigen administration. In pigs sensitized with serum in Freund's complete adjuvant HTA appeared at day 18 after the first antigen administration. Time course of appearance of HTA was about 10 days in both groups. Alum precipitated serum elicited considerably higher titres of HTA than equine serum in Freund's complete adjuvant. The passive cutaneous anaphylaxis activity of the HTA sera was located in the 7S fraction.