Effect of repeated fasting/refeeding on obesity development and health complications in rats arising from reduced nest.

BACKGROUND: Overnutrition during postnatal life represents a risk factor for later obesity and associated metabolic disorders. AIM: We investigated the interaction between postnatal and later-life nutrition on body composition, blood pressure and the jejunal enzyme activities in male Sprague-Dawley rats. METHODS: From birth, we adjusted the number of pups in the nest to 4 (small litters-SL; overfeeding) or to 10 pups (normal litters-NL; controls), and from day 50 until 70, the SL (SL-R) and NL (NL-R) rats were subjected to 1 day fasting and 1 day refeeding cycles (RFR). Their body composition was determined by magnetic resonance imaging, and enzyme activity was assayed histochemically. RESULTS: At 50 and 70 days, SL rats were found to be overweight (p < 0.001), with higher adiposity (p < 0.001) and blood pressure (p < 0.01). Moreover, despite significantly decreased daily food intake during RFR (SL-R 39 %, NL-R 23 %), higher fat deposition (p < 0.001) and blood pressure (p < 0.05) was detected in SL-R rats. Activity of alkaline phosphatase (AP) functionally involved in lipid absorption was significantly higher in SL than NL rats (p < 0.001) but substantially decreased in RFR groups (SL-R p < 0.001, NL-R p < 0.01). However, despite these enzymatic adaptations to reduced food intake, the SL-R rats displayed significantly higher AP activity in comparison with NL-R rats (p < 0.01) on day 70. CONCLUSIONS: Our results demonstrate that postnatal overfeeding predisposes the ontogeny of intestinal function, which may promote the probability of obesity risk. Accordingly, in these animals, efficient fat deposition and elevated blood pressure were not diminished in response to dietary restrictions in later life.

Long-term effect of altered nutrition induced by litter size manipulation and cross-fostering in suckling male rats on development of obesity risk and health complications.

OBJECTIVE: We investigated the long-term effect of pre-weaning nutrition on positive and/or adverse regulation of obesity risk and health complications in male Sprague-Dawley rats. DESIGN AND SUBJECT: Two experimental models were used in the present work: (1) To induce postnatal over- or normal nutrition, the litter size was adjusted to 4 (small litters-SL) and to 10 pups (normal litters-NL) in the nest, (2) in suckling pups at day 10, we used cross-fostering to identify the effect of altered dietary environment on their future body fat regulation, food intake, blood pressure, and the duodenal and jejunal alkaline phosphatase activity. After weaning, these control (NL, SL) and cross-fostered (NL-SL, SL-NL) groups were exposed to standard laboratory diet. RESULTS: On day 50, the SL in comparison with NL rats became heavier and displayed enhanced adiposity accompanied by significantly increased systolic blood pressure (19%) and duodenal (16%) and jejunal (21%) alkaline phosphatase (AP) activity. The impact of pre-weaning over-nutrition of NL-SL pups was associated with long-lasting positive effect on obesity. In contrast, SL-NL rats submitted until weaning to the opposite normalized feeding condition on day 50 showed significantly decreased fat deposition (21%), systolic blood pressure (20%), and AP activity in duodenum and jejunum (14%). CONCLUSIONS: These results contribute to a better understanding of how early-acquired dietary habits determine the attenuation or prevention of obesity development in later life and can provide some benefit for optimizing the future dietary strategies in young and adult obese individuals.

Effect of antibiotic treatment on intestinal microbial and enzymatic development in postnatally overfed obese rats.

OBJECTIVE: To investigate the effect of the microbiota-induced changes and early overfeeding after amoxicillin administration (a) in suckling pups via their dams up to 15 days of lactation and (b) in weaned pups on intestinal microbial/functional adaptability and obesity development in male Sprague-Dawley rats. DESIGN AND METHODS: Postnatal nutrition was elicited by adjusting the number of pups in the nest to 4 (small litters [SLs]) and 10 (normal litters [NLs]), while from days 21 to 40, both groups were fed with a standard diet. The numbers of Bacteroides/Prevotella (BAC) and Lactobacillus/Enterococcus (LAB) in the jejunum and colon were determined by fluorescence in situ hybridization technique, and jejunal alkaline phosphatase (AP), α-glucosidase and aminopeptidase activity was assayed histochemically. RESULTS: On day 40, the SL in comparison with NL animals displayed excess weight/fat gain accompanied by higher LAB and lower numbers of BAC, and with permanently higher AP activity. Moreover, these acquired changes continued in SL vs. NL rats and were not influenced by antibiotic treatment, which induced significant decrease in the quantity of LAB and BAC. CONCLUSIONS: These findings highlight the role of early life overfeeding upon the gut microbial/functional ontogeny and allow to distinguish their potential involvement in later risk of obesity.

Effect of Atropa belladonna L. on skin wound healing: biomechanical and histological study in rats and in vitro study in keratinocytes, 3T3 fibroblasts, and human umbilical vein endothelial cells.

The effect of Atropa belladonna L. (AB) aqueous extract on skin wound healing was studied in male Sprague-Dawley rats subjected to two parallel full-thickness skin incisions on the back. Specimens for histological evaluation were collected on days 2 and 5 whereas for biomechanical testing, they were collected on day 5. In the in vitro study, a different concentration of AB extract was used to test the differentiation of keratinocytes using a panel of selected antibodies, proliferation, and cell survival of 3T3 fibroblasts and human umbilical vein endothelial cells using the MTT-assay. Results of the in vivo experiments showed in AB-treated wounds a shortened process of inflammation and accelerated collagen formation, as well as significantly increased wound stiffness as compared with control tissues. The in vitro examination showed that control keratinocytes were cytokeratin 19 free, while samples exposed to the highest AB extract concentration expressed CK19. Moreover, all concentrations were stimulatory to human umbilical vein endothelial cell proliferation. In addition, only the AB extract at the lowest tested concentration increased fibroblast growth, but higher concentrations decreased cell survival. In conclusion, our results indicate that the AB water extract positively affects early phases of skin wound healing in rats. However, the in vitro results on the inverse relation between the concentration of the AB extract and its effects on cell proliferation may be important for future research.

Effect of equal daily doses achieved by different power densities of low-level laser therapy at 635 nm on open skin wound healing in normal and corticosteroid-treated rats.

Optimal parameters of low-level laser therapy (LLLT) for wound healing are still discussed. Hence, our study was aimed to compare effects of different power densities of LLLT at 635 nm in rats. Four, round, full-thickness, skin wounds were made on the backs of 48 rats that were divided into two groups (non-steroid laser-treated and steroid laser-treated). Three wounds were stimulated daily with a diode laser (daily dose 5 J/cm(2)) each with different power density (1 mW/cm(2), 5 mW/cm(2), and 15 mW/cm(2)), whereas the fourth wound served as a control. Two days, 6 days, and 14 days after surgery, eight animals from each group were killed and samples were removed for histological evaluation. In the non-steroid laser-treated rats, significant acceleration of epithelization and collagen synthesis 2 days and 6 days after surgery was observed in stimulated wounds. In steroid laser-treated rats, 2 days and 14 days after surgery, a decreased leucocyte/macrophage ratio and a reduction in the area of granulation tissue were recorded, respectively. In conclusion, LLLT, by the method we used, improved wound healing in the non-steroid laser-treated rats, but it was useless after corticosteroid treatment.

Developmental changes of gut microflora and enzyme activity in rat pups exposed to fat-rich diet.

The aim of this study was to investigate the effect of a high-fat (HF)/energy diet on the intestinal microbiota, the alkaline phosphatase (AP) activity, and related parameters of growth and obesity during the suckling and weaning periods in male Sprague-Dawley rats. From birth, nutrition in suckling pups was manipulated by feeding rat dams either HF or a standard diet, and then after weaning, by exposure of experimental pups to the HF, and control rats to normal diet. On days 15, 20, 40 the numbers of 2 microbial groups, i.e., Bacteroides/Prevotella (BAC) and the Lactobacillus/Enterococcus (LAB) in the jejunum, were determined by fluorescent in situ hybridization technique, and the AP activity was assayed histochemically. During all investigated periods HF pups gained body fat more rapidly than control animals, but from weaning they displayed significantly stunted growth resulting in final body weight loss. Obesity in HF rats was also accompanied by higher LAB and lower numbers of BAC and with permanently higher AP activity. Correlation of these data showed significant negative correlation between LAB, AP, and weight gain and energy efficiency, and significant positive correlation of BAC and AP activity with body fat. These data support the concept that postnatal nutritional experience represents an important factor affecting the ontogeny of intestinal microbial communities and intestinal function. These acquired changes could be a component of regulatory mechanisms involved in adverse and/or positive consequences of HF diet for adiposity, body weight, and energy-balance control in later life.

Postsurgical administration of estradiol benzoate decreases tensile strength of healing skin wounds in ovariectomized rats.

BACKGROUND: The most effective method of increasing the level of estrogen in the wounds of post-menopausal women undergoing routine surgical procedures is by long-term preoperative administration. However, in the case of acute surgery or trauma, the most effective method of increasing the level of estrogen is administration immediately pre- or postsurgery. This study, therefore, was aimed at assessing the effect of postsurgical administration of estradiol benzoate on wound healing in ovariectomized (OVX) Sprague Dawley rats. MATERIALS AND METHODS: Three months prior to the wound healing experiment, 16 rats were anesthetized and underwent ovariectomy, while the other eight rats were sham operated. Two parallel full thickness skin incisions and two round full thickness skin excisions were performed on the dorsum of each rat. Dose of 10 microg/d of estradiol benzoate was administered to eight OVX rats for 6 d postoperatively, whereas the other animals received a placebo. After 6 d, all animals were sacrificed and samples removed for biomechanical and histological evaluation. RESULTS: The mean wound tensile strength of OVX estrogen treated rats (9.54 +/- 3.24 g/mm(2)) was significantly lower compared with vehicle-treated OVX animals (14.57 +/- 4.12 g/mm(2)) as well as with control rats subjected to sham-OVX surgery (11.71 +/- 3.33 g/mm(2)). Nevertheless, the histological evaluation in OVX estrogen treated rats showed a significantly increased process of neo-angiogenesis associated with slightly decreased collagen deposition. CONCLUSION: Our results indicate that the question of the clinical significance of this type of hormone replacement therapy remains open and requires further research.

Experimental study on predicting skin flap necrosis by fluorescence in the FAD and NADH bands during surgery.

The objective of the present study was to assess the feasibility of using endogenous fluorescence in intraoperative prediction of skin flap necrosis. The investigation was carried out in 10 Sprague-Dawley rats in which proximally based over-dimensioned random pattern skin flaps were dissected on the back and thereafter fixed into position. Immediately after surgery on each rat, synchronous fluorescence spectra (Deltalambda=90 nm) from five parts of the skin flap surface were measured. The presence of necrosis was evaluated on postoperative day 7. In flap parts designated as necrotic (n=18), a significantly lower (P<0.001) fluorescent signal from the nicotinamide adenine dinucleotide (NADH) and flavin adenine dinucleotide (FAD) bands (360-380 and 440-460 nm, respectively) was detected in comparison with the vital parts (n=32) (for FAD:1767+/-39 versus 2579+/-65 auxiliary units [A.U]. and for NADH:11724+/-340 versus 16810+/-473 A.U.). The results suggested a close relationship between the fluorescent signals from the FAD and NADH bands on one side and flap necrosis or survival on the other side. Thus, the use of fluorescence spectroscopy may be considered as a valuable noninvasive tool for the prediction of skin flap necrosis.

Histological assessment of the effect of laser irradiation on skin wound healing in rats.

OBJECTIVE: The purpose of this study was to evaluate, from the histological point of view, the effect of diode laser irradiation on skin wound healing in Sprague-Dawley rats. BACKGROUND DATA: Various biological effects have been described in different studies after low-level laser therapy (LLLT). METHODS: Two parallel full-thickness skin incisions were performed on the back of each rat (n = 49) and immediately sutured. After surgery, one wound of each rat was exposed to laser irradiation (continuous mode, 670 nm, daily dose 30 J/cm2), whereas the parallel wound was not irradiated and served as control. Both wounds were removed 24, 48, 72, 96, 120, 144, and 168 h after surgery and routinely fixed and embedded in paraffin sections, stained with hematoxylin and eosin, van Gieson, periodic acid Schiff + periodic acid Schiff diastase, Mallory's phosphotungstic hematoxylin, and azur and eosin, and histopathologically evaluated. RESULTS: As compared to nonirradiated control wounds, laser stimulation shortened the inflammatory phase as well as accelerated the proliferative and maturation phase, and positively stimulated the regeneration of injured epidermis and the reparation of injured striated muscle. CONCLUSION: LLLT at 670 nm positively influences all phases of rat skin wound healing.

In vivo monitoring the changes of interstitial pH and FAD/NADH ratio by fluorescence spectroscopy in healing skin wounds.

The aim of our study was to evaluate the changes of interstitial pH and flavin adenine dinucleotide (FAD)/reduced nicotinamide adenine dinucleotide (NADH) ratio in healing skin wounds using fluorescence spectroscopy in Sprague Dawley rats. In the experiment, excisional and incisional models of wound healing were used. The florescein as the pH-sensitive probe using excitation spectra (lambda(Em) = 535 nm) was used for the measurement of pH changes, and synchronous fluorescence spectra (Deltalambda = 60 nm) for the monitoring of FAD/NADH ratio changes were measured from the surfaces of healing wounds. Increase of interstitial pH and FAD/NADH ratio was recorded during the time interval from the 15th to the 65th minute after surgery. The decrease of pH between the 48th and the 72nd hour after surgery as well as the increase of FAD/NADH ratio between the 72nd and the 96th hour of wound healing were recorded. The results indicate that the use of fluorescence spectroscopy may be considered as a valuable tool for noninvasive in vivo monitoring of selected redox parameters in the early phases of wound healing.