Muscle reorganisation through local injection of stem cells in the diaphragm of mdx mice

Background

The diaphragm is the major respiratory muscle affected by Duchenne muscular dystrophy (DMD) and is responsible for causing 80% of deaths. The use of mechanical forces that act on the body or intermittent pressure on the airways improves the quality of life of patients but does not prevent the progression of respiratory failure. Thus, diseases that require tissue repair, such as DMD, represent a group of pathologies that have great potential for cell therapy. The application of stem cells directly into the diaphragm instead of systemic application can reduce cell migration to other affected areas and increase the chances of muscle reorganisation. The mdx mouse is a suitable animal model for this research because its diaphragmatic phenotype is similar to human DMD. Therefore, the aim of this study was to assess the potential cell implantation in the diaphragm muscle after the xenotransplantation of stem cells.

Methods

A total of 9 mice, including 3 control BALB/Cmice, 3 5-month-old mdx mice without stem cell injections and 3 mdx mice injected with stem cells, were used. The animals injected with stem cells underwent laparoscopy so that stem cells from GFP-labelled rabbit olfactory epithelium could be locally injected into the diaphragm muscle. After 8 days, all animals were euthanised, and the diaphragm muscle was dissected and subjected to histological and immunohistochemical analyses.

Results

Both the fresh diaphragm tissue and immunohistochemical analyses showed immunopositive GFP labelling of some of the cells and immunonegativity of myoblast bundles. In the histological analysis, we observed a reduction in the inflammatory infiltrate as well as the presence of a few peripheral nuclei and myoblast bundles.

Conclusion

We were able to implant stem cells into the diaphragm via local injection, which promoted moderate muscle reorganisation. The presence of myoblast bundles cannot be attributed to stem cell incorporation because there was no immunopositive labelling in this structure. It is believed that the formation of the bundles may have been stimulated by cellular signalling mechanisms that have not yet been elucidated.     

Changing trends in mastitis

The global dairy industry, the predominant pathogens causing mastitis, our understanding of mastitis pathogens and the host response to intramammary infection are changing rapidly. This paper aims to discuss changes in each of these aspects. Globalisation, energy demands, human population growth and climate change all affect the dairy industry. In many western countries, control programs for contagious mastitis have been in place for decades, resulting in a decrease in occurrence of Streptococcus agalactiae and Staphylococcus aureus mastitis and an increase in the relative impact of Streptococcus uberis and Escherichia coli mastitis. In some countries, Klebsiella spp. or Streptococcus dysgalactiae are appearing as important causes of mastitis. Differences between countries in legislation, veterinary and laboratory services and farmers' management practices affect the distribution and impact of mastitis pathogens. For pathogens that have traditionally been categorised as contagious, strain adaptation to human and bovine hosts has been recognised. For pathogens that are often categorised as environmental, strains causing transient and chronic infections are distinguished. The genetic basis underlying host adaptation and mechanisms of infection is being unravelled. Genomic information on pathogens and their hosts and improved knowledge of the host's innate and acquired immune responses to intramammary infections provide opportunities to expand our understanding of bovine mastitis. These developments will undoubtedly contribute to novel approaches to mastitis diagnostics and control.     

The use of melatonin and progestagen–eCG to initiate reproductive activity in prepuberal Awassi ewe lambs

This experiment was conducted to evaluate the effect of administering hormonal treatments (melatonin and progestagen/equine chronic gonadotropin (eCG)) on advancing puberty in Awassi ewe lambs. Fifty-one 6-month-old ewe lambs of similar body weights (around 28 kg) were randomly assigned into four treatment groups; control (CON; n?=?14), melatonin (MEL; n?=?13), melatonin plus progestagen–eCG (MELPP; n?=?11), and progestagen–eCG (PP; n?=?13). Ewe lambs in the PP and MELPP groups were fitted with intravaginal progestagen sponges containing 60 mg medroxyprogesterone acetate for 14 days; 400 IU eCG were administered to each of these ewe lambs on the day of sponge removal. Ewe lambs in the MEL and MELPP groups received subcutaneous melatonin implants (Regulin®, 18 mg melatonin) 36 days before sponge insertion. Hormonal treatment had no effect on ewe lamb body weight change. Estrous behavior was greater (p?p?p?p?相似文献   

Diagnosis and treatment of presumptive pyelonephritis in an Asian elephant (Elephas maximus).

A 37-yr-old female Asian elephant (Elephas maximus) presented with anorexia, restlessness, and dark-colored urine. Urinalyses showed hematuria, leukocyturia, isosthenuria, proteinuria, granular casts, and no calcium oxalate crystals. Bloodwork revealed azotemia. Urine culture revealed a pure growth of Streptococcus zooepidemicus resistant to sulfamethoxazole-trimethoprim but susceptible to cephalosporins. A presumptive diagnosis of pyelonephritis was made based on bloodwork, urinalysis, and urine culture. The animal was treated with intravenous ceftiofur, and intravenous and per rectum fluids were given for hydration. The elephant's attitude and appetite returned to normal, the abnormal blood parameters resolved, and urinary calcium oxalate crystals reappeared after treatment, supporting presumptive diagnosis. Follow-up ultrasonography revealed an abnormal outline of both kidneys with parenchymal hyperechogenicity and multiple uterine leiomyomas.     

Wild ostrich (<Emphasis Type="Italic">Struthio camelus australis</Emphasis>) reproduction in Orbata,a nature reserve in Tunisia

The aim of the current investigation was to determine wild ostrich reproductive behaviour in Orbata Nature Reserve by observing 16 hens and 28 cocks over a seven-year period. Intense laying commenced in January, one month after the cessation of the rainy season, and 92% of the eggs were produced during the dry season (January to May, peaking in March). Over the seven years, 1,322 eggs were laid in 69 nests, which corresponded to an annual average production of 19.2 ± 9.1 eggs/nest and 11.8 eggs/hen. 24 nests (34.78%) were non-brooded, 17 nests (24.64%) were deserted in the course of incubation, and 28 nests (40.58%) possessed hatched eggs. All the non-incubated nests had egg losses equivalent to 46.6 ± 12.6%. Hatchability success of incubated eggs was 41.9 ± 12.0%. Ostriches tended to dig their nests adjacent to the reserve enclosure which had direct access by road and track, the latter subjecting them to human disturbance and predation. The systematic obstruction of these nests stimulated ostriches to build additional nests within the reserve perimeter. The authors discussed the results recorded in an ostrich flock in relation to the environmental factors (climatic factors, food disponibility and predation) and suggested possibilities for improved wildlife management.     

Quantification of pulmonary to systemic flow ratio by a Doppler echocardiographic method in the normal dog: Repeatability, reproducibility, and reference ranges

Objective

To determine if the pulmonary to systemic flow ratio (Qp/Qs) could be assessed in healthy awake dogs using Doppler echocardiography.

Background

Qp/Qs could provide reliable information in assessing the severity of intracardiac shunts (ICS) by quantifying pulmonary overcirculation. Qp/Qs has been validated against electromagnetic flowmeter methods in experimental canine models. However, its clinical applicability in awake dogs has never been assessed.

Animals, materials and methods

Six healthy dogs were used to determine the repeatability and reproducibility of the technique (Study 1); Qp/Qs was then prospectively assessed in 50 healthy dogs (Study 2). In both studies Qp/Qs was calculated in awake animals using a standardized Doppler echocardiographic method.

Results

Within- and between-day coefficients of variation for Qp/Qs were <10% (Study 1). For Study 2, a relatively wide range of Qp/Qs was found (reference range = 0.71-1.29; mean ± SD = 1.00 ± 0.15).

Conclusions

Qp/Qs can be assessed with good repeatability and reproducibility in healthy dogs. However, the wide range of Qp/Qs obtained in the healthy population may suggest a limited usefulness of this variable for accurately assessing ICS severity in diseased animals. This needs to be assessed in further prospective and longitudinal studies including a large number of animals with ICS of various grades.     

Effects of tea saponins on rumen microbiota,rumen fermentation,methane production and growth performance—a review

Reducing methane emission from ruminant animals has implications not only for global environmental protection but also for efficient animal production. Tea saponins (TS) extracted from seeds, leaves or roots of tea plant are pentacyclic triterpenes. They have a lasting antiprotozoal effect, but little effect on the methanogen population in sheep. There was no significant correlation between the protozoa counts and methanogens. The TS decreased methanogen activity. It seems that TS influenced the activity of the methanogens indirectly via the depressed ciliate protozoal population. The TS addition decreased fungal population in the medium containing rumen liquor in in vitro fermentation, but no such effect was observed in the rumen liquor of sheep fed TS. Tea saponins had a minor effect on the pattern of rumen fermentation and hence on nutrient digestion. When added at 3 g/day in diets, TS could improve daily weight gain and feed efficiency in goats. No positive associative effect existed between TS and disodium fumarate or soybean oil on methane suppression. Inclusion of TS in diets may be an effective way for improving feed efficiency in ruminants.     

Changes in nutrient balance,methane emissions,physiologic biomarkers,and production performance in goats fed different forage-to-concentrate ratios during lactation

The objective was to determine the effect forage-to-concentrate (F:C) ratio and stage of lactation on methane emissions, digestibility, nutrient balance, lactation performance, and metabolic responses in lactating goats. Twenty Murciano-Granadina dairy goats were used in an experiment divided into 3 periods: early (30 d), mid (100 d), and late (170 d) lactation. All goats were fed a diet with 35:65 F:C (FCL) during early-lactation. Then, 1 group (n = 10 goats) remained on FCL through mid- and late-lactation while the other group (n = 10 goats) was fed a diet with 50:50 F:C at mid-lactation (FCM) and 65:35 (FCH) at late lactation. A greater proportion of concentrate in the diet was associated with greater overall intake and digestibility (P < 0.05). Energy balance was negative in early-lactation (−77 kJ/kg of BW^0.75, on average) and positive for FCL at mid- and late-lactation (13 and 35 kJ/kg of BW^0.75, respectively). Goats fed FCM and FCH maintained negative energy balance throughout lactation. Plasma concentrations of non-esterified fatty acids at mid-lactation were greater for FCM than FCL (680 mEq/L), and at late-lactation concentrations were greater for FCH and FCL (856 mEq/L). A similar response was detected for plasma β-hydroxybutyrate. Methane emission was greater (P < 0.05) for FCM than FCH (1.7 g CH₄/d). This study demonstrated that differences in F:C across stages of lactation lead to distinct metabolic responses at the level of the rumen and tissues.     

Doxycycline treatment for <Emphasis Type="Italic">Dirofilaria immitis</Emphasis> in dogs: impact on <Emphasis Type="Italic">Staphylococcus aureus</Emphasis> and <Emphasis Type="Italic">Enterococcus</Emphasis> antimicrobial resistance

Doxycycline is an antibiotic that, in addition to the classic antibacterial use, is also prescribed to fight parasitic diseases, like heartworm disease in dogs. Despite the concern that the overuse of this antibiotic may decrease susceptibility of clinically important bacteria, the consequences of the prolonged doxycycline therapy in heartworm-infected dogs have never been studied before. We have analyzed the impact of this therapy on Staphylococcus aureus and Enterococcus antimicrobial resistance. In this study, 17 heartworm-infected dogs (10 that had completed the doxycycline treatment and 7 dogs that had not yet begun) were included. Twenty-four isolates of Staphylococcus aureus were obtained from two locations of each dog. After treatment, 73.3% of isolates were resistant to at least one antibiotic but only 22.2% of isolates before treatment. Most of doxycycline resistant isolates were obtained from dogs that have received treatment. Erythromycin resistance or intermediate susceptibility was detected in 45.6% of isolates, most of them from dogs after treatment. For Enterococci, 48 isolates were obtained from fecal samples (25 before treatment and 23 after treatment). Before treatment, 32% of isolates were resistant at least to one antibiotic while after, this data increase up to 65%. Comparing isolates before and after treatment, a clear increase in resistance to doxycycline (12% against 21.74%) and erythromycin (20% against 39.13%) was observed. Although the present work is a preliminary research, the results encourages the development of further studies to determinate the effect of prolonged doxycycline therapy on antimicrobial resistance.     

Tissue disposition of azithromycin after intravenous and intramuscular administration to rabbits

Tissue disposition of azithromycin after intravenous (IV) or intramuscular (IM) injection at a single dose rate of 10mg/kg bodyweight were investigated in rabbits using a modified agar diffusion bioassay for determining tissue concentrations. The pharmacokinetic behaviour of azithromycin was characterized by low and sustained plasma concentrations but high and persistent tissue concentrations. Kinetic parameters indicated a high retention of the drug in peripheral compartments. The plasma half-lives after IV and IM administrations were similar being 21.8h and 23.1h, respectively, while the half-lives obtained in tissues after IV and IM administration were at least 1.4 and 1.9 times longer than in plasma, respectively. The highest tissue concentrations were found in bile, liver and spleen whereas the lowest ones were found in skeletal muscle (although they were higher than those in plasma). From the results of the single administration in this study an IM dosage regimen can be proposed that achieves minimum concentrations over 2mg/L in rabbits: three doses of 4-5mg/kg/day would provide suitable therapeutic concentrations in pulmonary tissues over seven days.