Transgenerational effects of a hypercaloric diet.

The effects of a maternal hypercaloric diet (HD) during puberty and early adulthood on neuroimmune aspects in offspring were investigated. In female rats of the F0 generation and male rats of the F1 generation, bodyweight (BW) gain, retroperitoneal fat (RPF) weight, the number of hypodermic adipocytes (HAs) and expression of glial fibrillary acidic protein (GFAP) were measured in hypothalamic astrocytes. On Postnatal Day 50, the F1 pups were challenged with lipopolysaccharide (LPS, 100µgkg-1, s.c.) or an equal volume of saline (S), and behaviour in the open field test was evaluated, as were plasma neuropeptide and cytokine concentrations. The maternal HD caused the female F0 rats to become overweight. The F1 offspring of dams fed the HD and challenged with saline (HDS group) exhibited increases in BW gain, RPF weight and in the number of large HAs and a decrease in GFAP immunoreactivity. F1 offspring of dams fed the HD and challenged with LPS (HDLPS group) exhibited decreases in BW gain, RPF weight and GFAP immunoreactivity, but no differences were observed in the number of larger and small HAs. Plasma tumour necrosis factor-α concentrations were high in the HDS and HDLPS groups. Thus, the maternal HD during puberty and early adulthood caused the F1 generation to become overweight despite the fact that they received a normocaloric diet. These results indicate a transgenerational effect of the HD that may occur, in part, through permanent changes in immune system programming. The attenuation of neuroinflammation biomarkers after LPS administration may have resulted in a decrease in the number of adipocytes, which, in turn, reduced cytokine, adipokine and chemokine levels, which are able to recruit inflammatory cells in adipose tissue.

Maternal food restriction in rats of the F0 generation increases retroperitoneal fat, the number and size of adipocytes and induces periventricular astrogliosis in female F1 and male F2 generations.

The present study investigated whether male offspring (F2 generation) from female rats (F1 generation) whose mothers (F0 generation) were food restricted during gestation inherit a phenotypic transgenerational tendency towards being overweight and obese in the juvenile period, in the absence of food restriction in the F1/F2 generations. Dams of the F0 generation were 40% food restricted during pregnancy. Bodyweight, the number and size of larger and small hypodermal adipocytes (HAs), total retroperitoneal fat (RPF) weight and the expression of glial fibrillary acidic protein (GFAP) in periventricular hypothalamic astrocytes (PHAs), as determined by immunohistochemistry, were evaluated in both generations. In the female F1 generation, there was low bodyweight gain only during the juvenile period (30-65 days of age), a decrease in the size of small adipocytes, an increase in the number of small adipocytes, an increase in RPF weight and an increase in GFAP expression in PHAs at 90-95 days of age. In males of the F2 generation at 50 days of age, there was increased bodyweight and RPF weight, and a small number of adipocytes and GFAP expression in PHAs. These data indicate that the phenotypic transgenerational tendency towards being overweight and obese was observed in females (F1) from mothers (F0) that were prenatally food restricted was transmitted to their male offspring.

Ultra high dilution of triiodothyronine modifies cellular apoptosis in Rana catesbeiana tadpole tail in vitro.

BACKGROUND: Ultra High Dilutions (UHD) are diluted beyond the Avogadro limit with dynamization (dilution with succussion). The process of anuran amphibian metamorphosis is controlled by thyroid hormones, including the resorption of the tadpole tail. METHODS: A randomized and blinded study was performed to investigate the influence of triiodothyronine (T3) 5·10(-24)M (10cH) on apoptosis induced by T3 100 nM in Rana catesbeiana tadpoles' tail tips, in vitro. Explants were randomized to three groups: control: no T3 in pharmacological or UHD dose; test: T3 100 nM and challenged with T3 10cH (UHD); positive control: T3 100 nM, treated with unsuccussed ethanol. The apoptotic index and the area of explants of test and control groups at the first and final day of the experiment were compared by t-test. RESULTS: There was no difference in tail tip area between test and control groups, but a significantly higher (p<0.01) index of apoptosis in explants of the test group. CONCLUSION: This data suggest that T3 10cH modifies the effect of T3 at pharmacological dose, opening new perspectives for further studies and investigation of the dose-effect curve.

A randomized controlled trial of homeopathic treatment of weaned piglets in a commercial swine herd.

AIM: To evaluate the zootechnic performance and occurrence of diarrhea in piglets in the week post-weaning comparing supplementation with sucrose saline which contained or did not contain added homeopathic medicine. METHOD: Animals were randomly divided into three groups of 24 piglets each. The control group did not receive any treatment. Another group received sucrose saline, and the third group received sucrose saline with homeopathic medicine added, in the period of zero to seven days post-weaning. The homeopathic treatment consisted of Echinacea angustifolia, Avena sativa, Ignatia amara, Calcarea carbonica, all 6cH. Piglets were weighed daily for weight gain or loss, and observed for diarrhea and feed intake. RESULTS: Animals receiving sucrose saline alone and sucrose saline with homeopathy had less weight loss than control (p=0.017, p=0.0001 respectively). There was no statistical difference in relation to overall incidence of diarrhea or food consumption. These data suggest that the supplementation with sucrose saline with added homeopathic medicine in the first seven days post-weaning may be an useful option to reduce weight loss in weaned piglets.

Food deprivation in F0 generation and hypercaloric diet in F1 generation reduce F2 generation astrogliosis in several brain areas after immune challenge.

AIMS: The effects of maternal food restriction during gestation in F0 generation followed by hypercaloric diet (HD) during puberty in F1 generation (F1HD) were investigated on astrocyte behavior of F2 generation. Also, the astrocyte behavior, after an immune challenge, was examined by the immunohistochemical expression of glial fibrillary acidic protein (GFAP) in several brain areas. METHODS: The body weight gain (BW) during development and in postnatal day (PND) 90-95, the retroperitoneal fat weight (RPF), and the size of larger and smaller adipocytes in the F1 generation were assessed to observe the effects of HD in female rats. The BW, RPF weight and size of smaller and larger adipocytes was also measured to evaluate the transgenerational effects of F0 and F1 diets on F2 generation, treated or not with lipopolysaccharide (LPS). KEY FINDINGS: The F1HD group exhibited a higher BW gain than the F1 treated with normocaloric diet (ND, group F1ND), from weaning to PND65. In the frontal/parietal cortex, nucleus accumbens, hypothalamic arcuate/periventricular nuclei, molecular/granular layers of the cerebellum areas, excepting the pons, GFAP expression was greater in F1HD group relative to F1ND group. A reduced GFAP expression was observed in both groups born from F1 generation fed with HD (groups F2HDS and F2HDLPS) in relation to F2 generation born from dams fed with ND (groups F2NDS and F2NDLPS), independently of LPS challenge. SIGNIFICANCE: These data show an attenuation of LPS effect on GFAP expression, probably by a transgenerational effect of both maternal food deprivation in F0 generation and HD in F1 generation.

Silicea terra200cH evaluated by two different spectroscopy methods: a pilot study

Spectroscopy has been shown to bea useful method to study the physicochemical properties of homeopathic preparations. Aim: In this pilot study, we comparedtwo methods (photon scattering and visible-light spectroscopy) in the physicalevaluation of Silicea terra200cH. Methodology: Italian test: Two sampleshave been examinedand compared: Silicea terra200cH and Sac lac200cH, both preparedin water solution, diluted and succussed according to German Homeopathic Pharmacopoeia(Cemon Lab, Italy).Lactose was includedbecause the first 3 potencies of Silicea terraaremadeby trituration in lactosepowder.Measurements were made using an innovative Charge-Coupled Device (CCD) camera developedby Daniele Gullà, called MIRA/CORA(proprietary name). Slight variations in chrominance and luminance due to micro-vibrational 3D phenomenawere analyzed.Allmeasurements have been performed in a dark room at a constant temperature of 22°C +/-0,2°C usingafilter with very narrow spectral bands (10 nm).The measurements have been repeated three times on both Silicea terra200cH and Sac lac200cHwithin a few minutes aftereach other.Brazilian test: In the second test, performed in Brazil, variations in absorbance were used to identify Silicea terra200cH compared with Sac lac200cH andcompared withanother control solution of non-succussed 30% alcohol, using six solvatochromic dyes, following the method developed by Cartwright [1,2]. Both homeopathic samples were imported from Italy(the same sample bottles used in the Italian test),in Brazil they have beendiluted 1:100 in 30% hydro-alcoholic solution, and submitted to 100 succussions using an automatic mechanical arm (Denise, AUTIC, Brazil) prior to being tested. Samples were inserted into dyes solutions and evaluated by visible spectroscopy (FEMTO Spectrophotometer, Brazil). Dyes were prepared in ethanol P.A., according to previous established methods [3]. Three series in triplicate were performed and the results were analyzed by ANOVA / Tukey, comparing both samples and the unsuccussed 30% hydroalcoholic control solution.Results: Italian test: Measurements of the mean entropy of the signals, statistically elaborated with T Student test,yielded a two tailed p value < 0.05, where the entropy of the signal recorded from the Silicea terra200cHsample was statistically lower than the 200cH Sac lacsample.Brazilian test: Among all tested dyes, only BDN(4-(Bis-(4-(dimethylamino)-phenyl)methylene)-1(4H)-naphthalenone) showed aninteraction with Silicea terra200cH, reproducing the conclusions obtained in [3].Conclusions: Two different spectroscopic methods were able to show signal differences betweenSilicea terra200cH andSac lac200cH, suggesting that changes in solvent organization could be involved in the homeopathic signaling process, along withchanges in dipole moments of solvent and dyes. The results are potentially in line witha recent publishedpaper [4], that supportsthe propositionthatthe lower entropy of the verum signal compared with controls could beexplained by increased coherent vibrations of the verum sample, as theoretically explained in previous literature [5].

Hypercaloric diet prevents sexual impairment induced by maternal food restriction.

Prenatal undernutrition impairs copulatory behavior and increases the tendency to become obese/overweight, which also reduces sexual behavior. Re-feeding rats prenatally undernourished with a normocaloric diet can restore their physiological conditions and copulatory behavior. Thus, the present study investigated whether a hypercaloric diet that is administered in rats during the juvenile period prevents sexual impairments that are caused by maternal food restriction and the tendency to become overweight/obese. Female rats were prenatally fed a 40% restricted diet from gestational day 2 to 18. The pups received a hypercaloric diet from postnatal day (PND) 23 to PND65 (food restricted hypercaloric [FRH] group) or laboratory chow (food restricted control [FRC] group). Pups from non-food-restricted dams received laboratory chow during the entire experiment (non-food-restricted [NFR] group). During the juvenile period and adulthood, body weight gain was evaluated weekly. The day of balanopreputial separation, sexual behavior, sexual organ weight, hypodermal adiposity, striatal dopamine and serotonin, serum testosterone, and tumor necrosis factor α (TNF-α) were evaluated. The FRH group exhibited an increase in body weight on PND58 and PND65. The FRC group exhibited an increase in the latency to the first mount and intromission and an increase in serum TNF-α levels but a reduction of dopaminergic activity. The hypercaloric diet reversed all of these effects but increased adiposity. We concluded that the hypercaloric diet administered during the juvenile period attenuated reproductive impairments that were induced by maternal food restriction through increases in the energy expenditure but not the tendency to become overweight/obese.

Long-term accumulation and microdistribution of uranium in the bone and marrow of beagle dog.

The accumulation and microdistribution of uranium in the bone and marrow of Beagle dogs were determined by both neutron activation and neutron-fission analysis. The experiment started immediately after the weaning period, lasting till maturity. Two animal groups were fed daily with uranyl nitrate at concentrations of 20 and 100 microg g(-1) food. Of the two measuring techniques, uranium accumulated along the marrow as much as in the bone, contrary to the results obtained with single, acute doses. The role played by this finding for the evaluation of radiobiological long-term risks is discussed. It was demonstrated, by means of a biokinetical approach, that the long-term accumulation of uranium in bone and marrow could be described by a piling up of single dose daily incorporation.

The influence of social isolation and peripheral innervation on Ehrlich tumor growth in mice.

The relationship between social isolation and Ehrlich tumor growth was investigated in seven male NIH mice about 2 months old living separately in small cages for 28 days. Fifteen control animals were kept grouped in conventional cages (10 animals in one and 5 in another) for the same period. After this period, 40 to 50 microliters of a cell tumor suspension at a concentration of 1 to 5 x 10(7) tumor cells/ml was inoculated into the footpad and footpad size was measured for 10 days. Isolated mice presented a 30% reduction in tumor growth. Sciatic and saphenous neurectomy in one leg of 5 isolated mice (experimental) and 5 grouped mice (control) performed 7 days before tumor inoculation abolished this difference, and more tumor growth was observed in the neurectomized paw compared to the non-neurectomized paw. The subordinate x dominant social relationship established between mice living in groups of two per cage (seven cages) did not modify the growth of tumor inoculated under the same conditions compared to the first experiment. We conclude that social isolation and an intact peripheral innervation are associated with reduced tumor growth, but dominance behavior has no effect.

Substance P does not modify mononuclear cell migration into Ehrlich tumor mass.

In the present study, seven adult male mice were inoculated with Ehrlich tumor into the footpad after local substance P release was blocked by neurectomy of the sciatic and saphenous nerves. The contralateral footpad was also inoculated but sham-operated, and used as control. This procedure did not modify the percent of CD4+ (about 1-2%), CD8+ (about 1-3%), macrophages (about 21-22%), lymphocyte B (about 0-1%) and NK (about 1-2%) mononuclear cells present among tumor cells. These data suggest that chemotactic activity of substance P may not be relevant in this situation because the lack of this neurotransmitter (checked by immunohistochemistry) secondary to neurectomy did not change the cell migration profile into tumor mass.

Effect of a submaxillary gland extract on Ehrlich tumor growth in mice.

Ablation of host submaxillary glands modifies Ehrlich tumor growth and tumor-infiltrating leukocytes, possibly by modifications in the serum level of growth factors produced by this gland. To extend this research, 7-month-old male EPM-1 mice (N = 30) were divided into two groups: 1) inoculated with tumor cells previously incubated with submaxillary salivary gland extract (SGE) in PBS for 30 min at 37%; 2) inoculated with tumor cells previously incubated with PBS, under the same conditions. Animals were inoculated into the footpad with 40 microl of a suspension containing 4.5 x 10(7) tumor cells/ml, and footpad thickness was measured daily for 10 days. Sections and smears of tumor cells were prepared from the tumor mass to determine mitosis frequency, percent of tumor cells immunopositive to nerve (NGF) and epidermal (EGF) growth factors and percent of tumor-infiltrating leukocytes. The incubation of tumor cells with SGE produced a tumor reduction of about 30% in size (P<0.01). This effect was not related to loss of cell viability during incubation, but a 33% increase (P<0.05) in the percentage of dead or dying tumor cells and a 15% increase in the percent of NGF/EGF-positive tumor cells (P<0.01) were observed in vivo at the end of experiment. Tumor-infiltrating lymphocytes and mitosis frequency did not differ between groups. These data suggest a direct effect of factors present in SGE on tumor cells, which induce degeneration of tumor cells.

Therapeutic and pathogenetic animal models for Dolichos pruriens.

The therapeutic and pathogenetic effects of Dolichos pruriens were evaluated using experimental models in rats. In the therapeutic experiment Wistar rats were housed in a heated environment (25+/-3 degrees C) to induce itch, and treated with ascending potencies D. pruriens (6 cH, 9 cH, 12 cH and 30 cH), each for 10 days. The positive control group received vehicle (ethanol 30% in water). The negative control group received no treatment and were kept at a standard temperature. In the pathogenetic experiment, all animals were kept at a temperature of 20+/-3 degrees C and treated for 30 consecutive days with D. pruriens 6 or 30 cH, or ethanol vehicle, or no treatment. The experiments were performed blind. The statistical analysis used Bartlett's test, followed by ANOVA/Tuckey-Krammer or Kruskal-Wallis/Dunn. The results point to the existence of therapeutic effects, with inhibition of the itching, skin lesions and fur thinning produced by heat, more evident in later observations, with the 9 12, and 30 cH potencies (Kruskal-Wallis/Dunn; P=0.001). No changes were observed in the other parameters, such as open field activity and laterality of the itching. In the pathogenetic experiment, no changes were observed in any parameters examined. We conclude that the proposed experimental model demonstrates the therapeutic effect of D. pruriens, but not its pathogenetic effects.

Coccidiose clínica em frangos de corte infectados naturalmente e imunossuprimidos com dexametasona / Clinical coccidiosis in broiler chicks naturally infected and immunosuppressed with dexamethasone

A ocorrência de coccidiose clínica em aves está relacionada com a competência do sistema imune. Com o objetivo de avaliar a ocorrência natural de coccidiose em aves imunossuprimidas, foram selecionados frangos de corte, de ambos os sexos, com a 35 a 38 dias de vida para constituir 3 grupos - grupo I (n = 25), formado por aves sem coccidiose e negativas para coccídias no exame de fezes; grupo II (n = 25), formado por aves com coccidiose e positivas para coccídias no exame de fezes, e grupo III (n = 25), formado por aves sem coccidiose, negativas para coccídias no exame de fezes e submetidas à imunossupressão com dexametasona (4 mg/kg/dia por 4 dias, via subcutânea). Realizou-se o diagnóstico de coccidiose com a técnica de centrífugo-flutuação com solução saturada de sacarose para investigação de oocistos nas fezes e pela análise macro e microscópica das lesões intestinais observadas após a necropsia. A resposta imune foi avaliada pela reação de hipersensibilidade basofílica cutânea (CBH) à fitoemaglutinina (PHA) e pela relação entre o peso corporal e o peso da bursa de Fabricius ou do baço. Os frangos dos grupos II e III apresentaram menor reação CBH à PHA que os do grupo I, evidenciando-se diminuição da resposta imune. As aves do grupo III mostraram diminuição significante do peso da bursa de Fabricius e do baço em relação aos animais dos outros grupos. As espécies de coccídias encontradas foram E. acervulina e E. maxima nos animais dos grupos II e III, sendo ainda observada E. tenella nas aves do grupo III. A imunossupressão induzida pela dexametasona aumentou a suscetibilidade à coccidiose de ocorrência natural em frangos de corte criados comercialmente.

The purpose of this study was to evaluate the occurrence of clinical coccidiosis in broilers immunosuppressed with dexamethasone. Male and female broiler chickens, from 35 to 38 days old were divided into 3 groups ­ group I (n = 25), including chickens without coccidiosis and negative for coccidia in fecal examination; group II (n = 25), including birds with coccidiosis and positive for coccidia in fecal examination; group III (n = 25), constituted by chickens with no coccidiosis, negative for coccidia in fecal examination and immunosuppressed with dexamethasone (4 mg/kg/day for 4 days, subcutaneous route). The diagnosis of coccidiosis was achieved using the centrifugal floatation technique in sucrose solution to investigate the presence of oocystis in stools, as well as by the observation of macroscopic and microscopic changes in the gut after necropsy. The immune response was evaluated by determination of cutaneous basophil hypersensitivity (CBH) response to phytohemagglutinin (PHA) and of the weight ratio of bursa of Fabricius and spleen in relation to body weight. Broilers from group II and III presented decreased CBH reaction to PHA in relation to group I, suggesting a decrease of the immune response. In addition, chickens from group III presented a significant decrease in the weight of the bursa of Fabricius and of the spleen. The coccidian types were E. acervulina and E. maxima in chickens from groups II and III, as well as E. tenella in chickens treated with dexamethasone. Immunosuppression induced by dexamethasone increased susceptibility to natural coccidiosis in commercially raised broiler chicks.

Very high dilutions of dexamethasone inhibit its pharmacological effects in vivo.

We evaluated the interaction of dexamethasone 10(-17) and 10(-33) M (equivalent to 7cH and 15cH) with dexamethasone in pharmacological concentrations, using as experimental models: acute inflammation induced by carrageenan, Ehrlich ascitic tumour, and migration of tumour infiltrating leukocytes (TIL). Male adult BALB/c mice (n=7 per group) were used in all experiments. Carrageenan (1%) was injected into the footpad for oedema evaluation and into the peritoneal cavity (i.p.), for differential counting of inflammatory cells. Ehrlich ascitic tumour cells (10(7) viable cells/ml) were injected i.p. and tumour cells were counted after 6 days, by the Trypan blue exclusion method. The differential TIL was counted using smears stained by hematoxylin-eosin. Treatments were made immediately after carrageenan inoculation or once a day, during Ehrlich tumour development, until the animals were killed. Animals were treated with the following preparations: (1) phosphate buffer saline (PBS) solution; (2) dexamethasone (0.5 mg/kg for inflammation model or 4mg/kg for tumour model) mixed with dexamethasone 7cH or 15cH; (3) dexamethasone (same doses) mixed in PBS. Homeopathic dexamethasone partially blocked the anti-inflammatory effect of pharmacological dexamethasone with regard to paw oedema (two-way ANOVA, P < 0.0008) and polymorphonuclear cell migration (chi2, P=0.0001). No important differences were observed between experimental and control groups, in relation to Ehrlich tumour cells viability or count, or bodyweight, but potentised dexamethasone restored control levels of TIL viability, compared to mice treated with pharmacological doses of dexamethasone (chi2, P< or = 0.001). The results demonstrate that a potentised substance may change its own pharmacological effects and suggest that ultradilutions effects act mostly on host response.

Effect of Atropa belladonna and Echinacea angustifolia in homeopathic dilution on experimental peritonitis.

Atropa belladonna and Echinacea angustifolia have been used in homeopathy as modulators of inflammatory processes, in simple potency or 'accord of potencies', as recommended by homotoxicology. We evaluated their effects on leukocyte migration and macrophage activity induced by experimental peritonitis in vivo. Mice were injected (i.p.) with LPS (1.0mg/kg) and treated (0.3ml/10g/day, s.c.) with different commercial forms of these medicines. Echinacea angustifolia D4--a simple potency preparation--and Belladonna Homaccord, Belladonna Injeel, Belladonna Injeel Forte, Echinacea Injeel and Echinacea Injeel Forte--all in 'accord of potencies'--were tested. The association of A. belladonna and E. angustifolia in 'accord of potencies' produced an increase of polymorphonuclear cell migration (Kruskal-Wallis, P = 0.03) and a decrease of mononuclear cell percentages (Kruskal-Wallis, P < or = 0.04), when compared with control, mainly in preparations containing low potencies. The proportion of degenerate leukocytes was lower in the treated groups, compared to a control group (P < or = 0.05). The treated groups showed increased phagocytosis (P < or = 0.05), mainly in preparations containing high potencies. Our results suggest that A. belladonna and E. angustifolia, when prepared in 'accord of potencies', modulate peritoneal inflammatory reaction and have a cytoprotective action on leukocytes.

Action of Causticum in inflammatory models.

The anti-inflammatory effect of Causticum was evaluated using acute and chronic inflammatory models in vivo. The administration of concentrated Causticum solution into the hind paw of rats produced an inflammatory reaction with oedema formation within the first hour, showing that Causticum acts as an oedematogenic agent. Carrageenin induced rat paw oedema was significantly inhibited (P<0.05) in the group treated with Causticum 30cH solution compared to control. Groups treated with potentized Causticum (6cH, 12cH, 30cH and 200cH), showed significant inhibition (P<0.05) of the inflammation pre-induced by carrageenin. However pre-treatment with Causticum 30cH for 6 days (0.5 ml, daily) did not significantly inhibit granulation using an implantation method.

Effect of a submaxillary gland extract on Ehrlich tumor growth in mice

Ablation of host submaxillary glands modifies Ehrlich tumor growth and tumor-infiltrating leukocytes, possibly by modifications in the serum level of growth factors produced by this gland. To extend this research, 7-month-old male EPM-1 mice (N = 30) were divided into two groups: 1) inoculated with tumor cells previously incubated with submaxillary salivary gland extract (SGE) in PBS for 30 min at 37 percent; 2) inoculated with tumor cells previously incubated with PBS, under the same conditions. Animals were inoculated into the footpad with 40 µl of a suspension containing 4.5 x 107 tumor cells/ml, and footpad thickness was measured daily for 10 days. Sections and smears of tumor cells were prepared from the tumor mass to determine mitosis frequency, percent of tumor cells immunopositive to nerve (NGF) and epidermal (EGF) growth factors and percent of tumor-infiltrating leukocytes. The incubation of tumor cells with SGE produced a tumor reduction of about 30 percent in size. This effect was not related to loss of cell viability during incubation, but a 33 percent increase 0.05 in the percentage of dead or dying tumor cells and a 15 percent increase in the percent of NGF/EGF-positive tumor cells 0.01 were observed in vivo at the end of experiment. Tumor-infiltrating lymphocytes and mitosis frequency did not differ between groups. These data suggest a direct effect of factors present in SGE on tumor cells, which induce degeneration of tumor cells

The influence of social isolation and peripheral innervation on Ehrlich tumor growth in mice

The relationship between social isolation and Ehrlicha tumor growth was investigated in seven male NIH mice about 2 months old living separately in small cages for 28 days. fifteen control animals were kept grouped in conventional cages (10 animals in one and 5 in another) for the same period. After this period, 40 to 50 µl of a cell tumor suspension at a concentration of 1 to 5 x 10**7 tumor cells/ml was incolulated into the footpad and footpad size was measured for 10 days. Isolated mice presented a 30 percent reduction in tumor growth. Sciatic and saphenous neurectomy in one leg of 5 isolated mic (experimental) and 5 grouped mice (control) performed 7 days before tumor inoculation abolished this difference, and more tumor growth was observed in the neurectomized paw compared to the non-neurectomized apw. The subordinate x dominant social relationship established between mice living in groups of two per cage (seven cages) did not modify the growth of tumor inoculated under the same conditions comapared to the first experiment. We conclude that social isolation and an intact peripheral innervation are associated with reduced tumor growth, but dominance behavior has no effect

Substance P does not modify mononuclear cell migration into Ehrlich tumor mass

In the present study, seven adult male mice were inoculated with Ehrlich tumor into the footpad after local substance P release was blocked by neurectomy of the sciatic and saphenous nerves. The contralateral footpad was also inoculated but sham-operated, and used as control. This procedure did not modify the percent of CD4+ (about 1-2 per cent), CD8+ (about 1-3 per cent), macrophages (about 21-22 per cent), lymphocyte B (about 0-1 per cent) and NK (about 1-2 per cent) mononuclear cells present among tumor cells. These data suggest that chemotactic activity of substance P may not be relevant in this situation because the lack of this neurotransmitter (checked by immunohistochemistry) secondary to neurectomy did not change the cell migration profile into tumor mass.