Changes In Clinical Signs Research Articles

Toxicologic evaluation of Dichrostachys glomerata extract: Subchronic study in rats and genotoxicity tests

In western Cameroon, edible fruits and seeds from the plant Dichrostachys glomerata are commonly used as spices. Extract from the fruit pods has been reported as a good natural source of antioxidants and may provide health benefits. The objective of the present study was to investigate potential adverse effects, if any, of D. glomerata fruit pod extract (Dyglomera™) in a subchronic toxicity study and in genotoxicity studies. In the toxicity study, Sprague Dawley rats (20/sex/group) were gavaged with D. glomerata extract at dose levels of 0, 100, 1000 and 2500mg/kg body weight (bw)/day for 90-days. Dyglomera™ administration did not result in mortality or show treatmentrelated changes in clinical signs of toxicity, body weights, body weight gain or feed consumption. Similarly, no toxicologically significant treatment-related changes in hematological, clinical chemistry, urine analysis parameters, and organ weights were noted. Macroscopic and microscopic examinations did not reveal treatment-related abnormalities. Mutagenic and clastogenic potentials as evaluated by Ames assay, in vitro and in vivo chromosomal aberration test and in vivo micronucleus test did not reveal any genotoxicity of the extract. The results of subchronic toxicity study supports the no-observed-adverse-effect level (NOAEL) for D. glomerata extract as 2500mg/kg bw/day, the highest dose tested.

Six months chronic toxicological evaluation of naringin in Sprague–Dawley rats

Naringin is a flavonoid showing variable pharmacological properties and is distributed ubiquitously in plant foods. There is a paucity of reported data regarding its safety profile. In the present study, chronic toxicity studies of naringin was designed and conducted by oral gavage at doses of 0, 50, 250 and 1250mg/kg in Sprague–Dawley (SD) rats for six months followed by 1-month recovery period. During the 6-month treatment period and one month recovery period, no mortality and toxicologically significant changes in clinical signs, opthalmoscopic examination, hematology, clinical biochemistry, serumsexhormone, macroscopic findings, organ weights and histopathological examination were noted and attributed to naringin administration. Although consecutive and/or isolated periods of significant body weights and food consumption decreases were relevant to naringin administration, they were not considered toxicologically significant. In addition, slight, non-pathological and reversible hair loss was noted during the 6-month treatment period and considered as a kind of change possibly relevant to naringin administration; however, it was not considered adverse change and to be of toxicological significance. Based on the results of this study, the no-observed-adverse-effect-level (NOAEL) of naringin in rats is greater than 1250mg/kg/day when administered orally for 6 consecutive months.

Canine Oral Eosinophilic Granuloma Treated with Electrochemotherapy

A case of a canine oral eosinophilic granuloma in a 14-year-old female crossbred is described. The dog was presented with a history of ptyalism, halitosis, local pain, decreased appetite, and blood staining noted on food and water bowls. Clinical, hematologic, and biochemical examinations, abdominal ultrasonography, and 3-view chest radiographs were performed, and no metastases were found. Histopathologic examination of two 6 mm punch biopsies from the oral lesion revealed the presence of eosinophilic granulomatous lesions in the submucosa. After treatment with corticosteroids and wide spectrum antibiotics no significant changes in clinical signs and lesion size were observed. Electrochemotherapy (ECT), a novel tumor treatment routinely used for cutaneous and subcutaneous tumors in human patients in the European Union since 2006, was used to treat the eosinophilic granuloma. The procedure was performed under general anesthesia, followed by intravenous administration of bleomycin. Six weeks after treatment a complete response with disappearance of the mass and improvement of clinical signs were observed.

Subacute Oral Toxicity of Yukmijiwhang-Tang in Crl:CD Sprague-Dawley Rats and Its Cytotoxicity.

Background. The traditional herbal formula Yukmijiwhang-tang (YMJ) consists of six medicinal herbs and has been used to treat dysuria, diabetic mellitus, and neurosis in Korea, China, and Japan. Here we report safety information on its subacute toxicity and the cytotoxicity. Methods. YMJ extract was administered to SD rats at various dosages for 4 weeks. We monitored clinical signs, mortality, body and organ weights, food intake, and hematological and serum biochemistry factors. For cytotoxicity testing, each cell line was treated with various concentrations of YMJ for 24 h. Results. YMJ treatment had no significant effects on changes in clinical signs, body weight, or food intake in male or female rats. In male rats, YMJ treatment decreased the absolute weights of the epididymides and serum Na levels. In female rats, YMJ significantly reduced the prothrombin time (PT) and serum creatine level. However, the changes were not severe and were considered to be in the normal physiological range for rats. The no-observed-adverse-effect-level (NOAEL) was estimated to be 2000 mg/kg/day. YMJ extract did not exert any cytotoxicity against 23 tested cell lines. Conclusions. Our data provide scientific evidence on the safety of YMJ for potential development as a prescription drug.

Abstract P5-02-01: Regulated intratumoral expression of IL-12 in combination with cytotoxic agents as a strategy for the treatment of metastatic breast cancer

Abstract Major obstacles for the development of immunotherapeutics are the ability of tumors to escape the immune system coupled with toxicity associated with systemic administration. To overcome these challenges, we have developed an adenoviral vector, Ad-RTS-IL-12 (AD), administered intratumorally (IT) under control of the RheoSwitch Therapeutic System® (RTS) technology platform. Expression of IL-12 mRNA and IL-12 protein is tightly regulated by the oral administration of a small molecule activator ligand, veledimex (AL). We have previously demonstrated a concentration-related increase in IL-12 mRNA concomitant with increase in expression of IL-12 protein in HT1080 cells transduced with AD and incubated with AL. Removal of AL from the media resulted in a return to baseline IL-12 expression within 48 hours. Results from the subcutaneous 4T1 syngeneic BALB/c mouse mammary tumor model demonstrated an AL dose-related increase in tumor IL-12 mRNA and IL-12 protein expression with the maximum IL-12 tumor protein level of 280 ng/mg achieved at 150 mg/m2 AL + 1e10 vp AD. In addition, a return to baseline IL-12 mRNA and IL-12 protein expression was observed on cessation of AL. The effect of AD + AL on tumor growth rate was evaluated in a subcutaneous 4T1 syngeneic BALB/c mouse mammary tumor model. A single intratumoral injection of 1e10 vp AD combined with oral administration of AL (15, 30, 75 or 150 mg/m2) on a Q1Dx5 schedule led to significant AL dose-related tumor growth inhibition with tumor size reduction of 27%, 37%, 57% and 60%, respectively when compared to vehicle on Day 33. Neither AD or AL alone had an effect on tumor growth rate relative to vehicle control. No change in clinical signs or body weight was observed when compared to vehicle alone. Tumor growth inhibition correlated with an increase in tumor IL-12 levels, a decrease in Tregs, and an increase in cytotoxic T cells. Results from a phase 1 study in melanoma have shown dose-related production of IL-12 and clinical activity in injected and non-injected lesions at 100 and 160 mg of AL. Cytotoxic agents at low doses have been shown to prime the immune system and combination with immunotherapy may augment tumor specific T-cell immune responses resulting in enhanced efficacy. In the 4T1 mouse mammary tumor model Ad-RTS-mIL12 (1e10vp) + AL (30 mg/m2) combined with palifosfamide (40 or 120 mg/m2 QD IP for 3 days) significantly inhibited tumor growth (∼71-90% vs. control) concomitant with increased median survival when compared to the single agents alone. Based on these findings in a multicenter, open-label, randomized, phase 2 study evaluating the safety and efficacy of AD + AL alone or in combination with cytotoxic agents in subjects with recurrent/metastatic breast cancer with accessible tumor(s) is ongoing. This study assesses AD + AL administered as a monotherapy and a combination therapy. In the monotherapy arm, AD is administered IT on Day 1 and AL 140 mg is administered orally on either a Q1Dx7 or QODx14 schedule. In the combination arm, palifosfamide 120 mg/m2 IV Q1Dx3 is be administered with AD + AL QODx14 (28 day cycle). Preliminary data show that AD + AL is safe and well tolerated. Preclinical and clinical data will be presented. Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P5-02-01.

Strength gain through eccentric isotonic training without changes in clinical signs or blood markers

BackgroundLocalized exercises are widely used in rehabilitation processes. The predominant options are exercises with an emphasis on either concentric or eccentric contractions. Eccentric exercises promote greater strength gains compared to classical concentric stimuli, but can cause muscle damage. The aim of present study was to compare strength training composed of 10 sessions with progressive loads between groups with a predominance of concentric versus eccentric contraction through an analysis of isotonic strength, pressure pain threshold, creatine kinase, tumor necrosis factor-alpha and cortisol.MethodsOne hundred twenty male subjects were divided into four groups: C1 and E1 – single session of maximum strength with emphasis on concentric and eccentric contraction, respectively; C10 and E10 – 10 sessions with progressive loads from 80% to maximum strength with emphasis on concentric and eccentric contraction, respectively.ResultsIsotonic strength increased by 10% in E10 following the ten training sessions. C1 and E1 exhibited a lower pressure pain threshold 48 hours after the sessions in comparison to C10 and E10, respectively. Creatine kinase was increased in C1 in comparison to baseline, with significant differences (p ≤ 0.05) in comparison to E1 at 48 and 96 hours as well as C10 at 48, 72 and 96 hours. No significant differences were found in TNF-α or cortisol among the groups or evaluation times.ConclusionEccentric contraction training promotes functional adaptation. Moreover, both concentric and eccentric contraction training have a protective effect on the muscle in relation to a single session of maximum strength exercise.Trial registrationRBR-75scwh

Acute and subacute toxicity studies of Pluronic P85/poly (lactic acid) nanoparticles in mice

The objective of the present reported study was to evaluate the oral toxicity of poly (lactic acid)-b-Pluronic-b-poly (lactic acid) (PLA-P85-PLA) nanoparticles (NPs) so as to demonstrate their applicability for drug delivery applications. In acute oral toxicity studies, the animals were fed with 0.4 ml PLA-P85-PLA NPs solution in different concentrations (low concentration: 20 mg/l, intermediate concentration: 100 mg/l and higher concentration: 500 mg/l) for 14 days; no deaths or treatment related complications were observed even in the higher concentration treatment. In case of the subacute oral toxicity test, the NPs were administered orally to mice for a period of 28 days. At the end of the study, blood samples were collected for biochemical analysis. For histopathological analysis, organs of the animals were weighed and processed. All the animals survived during the study, with no significant changes in clinical signs, body weight, feed consumption, biochemistry parameters, organ weights and histopathological findings. These results demonstrate that PLA-P85-PLA NPs produced no treatment related toxicity in mice following oral administration, thus, they can be exploited for potential therapeutic applications.

Acute and 13weeks subchronic toxicological evaluation of naringin in Sprague-Dawley rats

Naringin is widely distributed in plant foods and has not previously been evaluated for safety through standard in vivo toxicological studies. In the present study, acute and subchronic oral toxicity studies of naringin were designed and conducted in Sprague-Dawley (SD) rats. Acute oral administration of naringin was done as a single bolus dose up to 16 g/kg and subchronic toxicity study for 13 weeks was done by oral administration at doses of 0 (control), 50, 250 and 1250 mg/kg in SD rats. There were no mortality, adverse clinical signs, abnormal changes in body weights or food consumption, toxicologically relevant changes in hematology, clinical biochemistry and macroscopic findings during 14 days of the acute toxicity study. During the subchronic oral toxicity study, no mortality and toxicologically significant changes in clinical signs, food consumption, opthalmoscopic examination, hematology, clinical biochemistry, serum sex hormone, macroscopic findings, organ weights and histopathological examination except for slight body weight decrease were noted and attributed to naringin administration. These observations suggest that naringin is practically non-toxic for SD rats in oral acute toxicity study and the no-observed-adverse-effect-level (NOAEL) of naringin in rats is greater than 1250 mg/kg/day when administered orally for 13 consecutive weeks.

OP0212 Very Long-Term Effects of the “4 Plus 2 Infusion Protocol” of Rituximab Alone in Patients with HCV-Associated Mixed Cryoglobulinemia with Diffuse Membranoproliferative Glomerulonephritis, Severe Polyneuropathy and Necrotic Ulcers of Skin

BackgroundMixed cryoglobulinemia syndrome (MCs) is a systemic vasculitis characterized by multiple organ involvement due to the vascular deposition of IgMk/IgG cryoglobulins. B cells expansion usually triggered by HCV infection plays...

Abstract 2838: Intratumoral-regulated expression of IL-12 as a potential immunotherapeutic strategy for the treatment of breast cancer.

Abstract A major obstacle for the development of immunotherapeutics is the ability of tumors to escape the immune system coupled with toxicity associated with systemic administration of cytokines. To overcome these challenges we have developed an adenoviral vector with regulated expression of IL-12, Ad-RTS-mIL-12 (Ad), was administered intratumorally under the control of the RheoSwitch Therapeutic System® (RTS®) expression platform. Previously we have shown that a single intratumoral injection of Ad combined with oral administration of the activator ligand, INXN-1001 (AL), led to significant inhibition of tumor growth and increased survival. Herein, we show that localized delivery of IL-12 is well tolerated and its mechanism of action is through an increase in tumor infiltrating leukocytes concomitant with a reduction in tumor growth. In vitro studies using HT1080 human fibrosarcoma cells transduced with Ad and incubated with AL have shown a concentration-related increase in IL-12 mRNA concomitant with an increase in expression of IL-12 protein. Removal of AL from the media resulted in return of IL-12 expression to baseline within 48 hours. In the 4T1 BALB/c mouse model, oral administration of AL elicited a dose- related increase in plasma AL levels which correlated with increasing tumor levels of AL. The increase in tumoral AL in turn activated the RheoSwitch® portion of the adenoviral vector leading to a dose-related increase in tumor IL-12 production with virtually no increase in systemic IL-12. The maximal tumoral IL-12 protein level of ∼280 pg/mg protein was achieved on Day 4 of AL dosing with a concomitant serum IL-12 level of 0.5 pg/mg protein following a single intratumoral injection of Ad-RTS-mIL-12 (1x1010 vp) on Day 1 combined with once daily oral administration of AL at 150 mg/m2 on Days 1-7. This increase in tumoral IL-12 levels correlated with a dose-related increase in tumor-infiltrating CD4-positive and CD-8 positive lymphocytes in and adjacent to the tumor, coupled with an increase in apoptotic marker caspase 3 in the tumor when compared with vehicle. No increase in IL-12 protein expression in mice treated with Ad or AL alone was observed. Moreover, this therapeutic strategy appears to be well-tolerated as no change in clinical signs or body weight was observed in the treated animals when compared with vehicle alone. These results support our hypothesis that localized delivery of IL-12 is well tolerated and results in an increase in tumor infiltrating leukocytes concomitant with a reduction in tumor growth. These findings suggest the applicability of our immunotherapeutic strategy for the treatment of metastatic breast cancer. Citation Format: John A. Barrett, Lei Sun, Maria Grigoriadis, Christan Fulan-Freguia, Tim Chan, Bill Fogler, Laurent Humeau, Roya Khosravi-Far, Robert A. Morgan, Hagop Youssoufian. Intratumoral-regulated expression of IL-12 as a potential immunotherapeutic strategy for the treatment of breast cancer. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 2838. doi:10.1158/1538-7445.AM2013-2838

Outcome of ventriculoperitoneal shunt implantation for treatment of congenital internal hydrocephalus in dogs and cats: 36 cases (2001–2009)

To examine outcome data for cats and dogs with congenital internal hydrocephalus following treatment via ventriculoperitoneal shunting to determine treatment-associated changes in neurologic signs, the nature and incidence of postoperative complications, and survival time. Retrospective multicenter case series. 30 dogs and 6 cats with congenital internal hydrocephalus (confirmed via CT or MRI). Medical records for dogs and cats with internal hydrocephalus that underwent unilateral ventriculoperitoneal shunt implantation from 2001 through 2009 were evaluated. Data collected included the nature and incidence of postoperative complications, change in clinical signs following surgery, and survival time. To compare pre- and postoperative signs, 2-way frequency tables were analyzed with a 1-sided exact McNemar test. 8 of 36 (22%) animals developed postoperative complications, including shunt malfunction, shunt infection, and seizure events. Three dogs underwent shunt revision surgery. Thirteen (36%) animals died as a result of hydrocephalus-related complications or were euthanized. Following shunt implantation, clinical signs resolved in 7 dogs and 2 cats; overall, 26 (72%) animals had an improvement of clinical signs. After 18 months, 20 animals were alive, and the longest follow-up period was 9.5 years. Most deaths and complications occurred in the first 3 months after shunt placement. Results indicated that ventriculoperitoneal shunt implantation is a viable option for treatment of dogs or cats with congenital hydrocephalus. Because complications are most likely to develop in the first 3 months after surgery, repeated neurologic and imaging evaluations are warranted during this period.

A 13-week subchronic toxicity study of grape skin extract in F344 rats

A 13-week repeated oral dose toxicity study of grape skin extract (GSE) was performed using F344 rats. Four groups of animals, each consisting of ten males and ten females, were fed a diet containing 0%, 0.2%, 1.0% or 5.0% GSE for 13 weeks. Throughout the experiment, there were no treatment-related changes in clinical signs, body weight or mean food intake in any of the treated groups of either gender. Hematological studies and serum biochemical analyses revealed no treatment-related changes in all groups in both genders. In the glandular epithelial cells of the parotid glands, diffuse hypertrophy and basophilia was observed in all animals in both 5.0% groups. Hypertrophy of the parotid glands was not detected in the 0.2% or the 1.0% dose groups. In female kidneys, slight calcification in the renal proximal tubules of the cortex and medulla was observed in all groups including controls. This is a common spontaneous change in female rats, and the incidence was comparable between controls and treated groups. However, the number of tubules with calcification was higher in the 5.0% group based on a semi-morphometric analysis. Based on the histopathology of the parotid glands andthe minor change in the kidneys, the no observed adverse effect level (NOAEL) of GSE in the present study was a 1.0% treatment dose in both genders (males: 0.6 ± 0.2 g/kg body weight/day; females: 0.7 ± 0.1 g/kg body weight/day).

Subchronic toxicity and mutagenicity/genotoxicity studies of Irvingia gabonensis extract (IGOB131)

African Bush Mango from Irvingia gabonensis is a West African culinary fruit and the mucilage from this fruit seed is used to make traditional soups and sauces. Extract from the kernel (IGOB131) has been claimed for its health benefits. In the present investigations, potential adverse effects, if any, of IGOB131 were investigated in dose–response 90-day study and genotoxicity studies. In the subchronic study, Sprague Dawley rats (20/sex/group) were gavaged with I. gabonensis extract (IGOB131) at dose levels of 0, 100, 1000 and 2500mg/kg body weight (bw)/day for 90-days. No treatment-related changes in clinical signs, functional observations, mortality, ophthalmologic observations, body weights, body weight gain or feed consumption were noted. Similarly, hematological, clinical chemistry, urine analysis parameters, and organ weights did not reveal any toxicologically significant treatment-related changes. No treatment-related macroscopic and microscopic abnormalities were noted at the end of treatment period. The mutagenicity as evaluated by Ames assay, in vitro and in vivo chromosomal aberration test and in vivo micronucleus assay did not reveal any genotoxicity of IGOB131. The results of subchronic toxicity study suggest the no-observed-adverse-effect level (NOAEL) for I. gabonensis extract (IGOB131) as ⩾2500mg/kg bw/day, the highest dose tested.

Refeeding Syndrome or Refeeding Hypophosphatemia

Nutrition support clinicians refer to the abnormalities in laboratory data and changes in clinical signs and symptoms that follow refeeding of starved or malnourished patients as refeeding syndrome. Theoretical descriptions of refeeding syndrome include a complex and extensive list of changes, such as hypophosphatemia, hypomagnesemia, hypokalemia, hyponatremia, hypocalcemia, hyperglycemia, and vitamin deficiency--all of which are accompanied by clinical signs and symptoms. In practice, clinicians see asymptomatic refeeding hypophosphatemia more often than a full-blown syndrome with multiple laboratory and clinical abnormalities. Confusion results because there is no widely accepted or uniformly applied set of defining characteristics for diagnosing refeeding syndrome. To gain insight into the clinical characteristics of refeeding syndrome described in the literature, a systematic review of reported cases and case series was conducted. Since 2000, 20 authors described 27 cases that contained sufficient data for review. Hypophosphatemia occurred in 26 patients (96%). While 19 patients (71%) experienced at least 1 other laboratory abnormality, only 14 (51%) exhibited a consistent pattern of abnormally low phosphorus and magnesium levels. Seven patients had hypocalcemia (26%), and hyponatremia was reported in 3 patients (11%). There were no reports of hyperglycemia. Mean data reported in case series containing data from 63 patients showed that hypophosphatemia was a consistent finding but that other abnormalities were not consistently identified. Findings suggest that refeeding hypophosphatemia is not accompanied by a consistent pattern of biochemical or clinical abnormalities among case reports or case series of patients reported to have refeeding syndrome.

Pulmonary and systemic responses of highly pure and well-dispersed single-wall carbon nanotubes after intratracheal instillation in rats

The present study was conducted to assess the pulmonary and systemic responses in rats after intratracheal instillation of highly pure, well-dispersed, and well-characterized SWCNTs. Exposure to SWCNTs up to 2 mg/kg did not produce mortality, changes in clinical signs, or body weights during the observation period. Dose-dependent changes were observed in the lung weight, BALF inflammatory cells, and biochemical parameters such as LDH value, protein content, IL-1β and IL-6 activity, and histopathology. In the 0.04 mg/kg SWCNT-exposed group, almost no changes were observed during the observation period. In the 0.2 mg/kg SWCNT-exposed group, pulmonary inflammatory responses were observed after instillation. In the 1 mg/kg and 2 mg/kg SWCNT-exposed group, acute lung inflammation and subsequent granuloma accompanied by increased lung weights were observed. Furthermore, the histopathological findings in the lungs of rats exposed to SWCNTs showed inflammatory responses related with the vital reaction to the foreign substance that was instilled intratracheally, and there were no fibrosis, atypical lesion, or tumor-related findings even at the highest dose (2 mg/kg) of SWCNT-exposed groups up to 6 months after instillation. For all groups, histopathological changes due to the instillation exposure of SWCNTs were observed only in the lungs and lung-associated lymph nodes and not in the other tissues examined (i.e. the liver, kidney, spleen, and cerebrum).

28-day repeated dose oral toxicity of human copper-zinc superoxide dismutase from recombinant Pichia pastori in rats

Human copper/zinc superoxide dismutase from recombinant Pichia pastori (RH-Cu/Zn-SOD) was orally administered, via gavage, to Sprague-Dawley rats at 500, 1,000, and 2,000 mg/kg body weight/day for 28 days. During the 28-day period, animals were examined for evidence of toxicity; there were no deaths, and in-life physical signs were normal. On day 29, the animals were exsanguinated, examined for gross pathology, and tissues were preserved for histopathology. Although statistical differences were noted in some hematology and clinical chemistry, they were of questionable biological significance. The results of the 28-day oral administration demonstrated a lack of toxicity of RH-Cu/Zn-SOD in rats. There were no treatment-related, toxicologically relevant changes in clinical signs, growth, food consumption, hematology, clinical chemistry, organ weights, or pathology. The no observed adverse effect level was greater than 2,000 mg/kg/day for RH-Cu/Zn-SOD in rats.

Acute and 90-day subchronic toxicity studies of Silk peptide E5K6, in Sprague–Dawley rats

Acute and 90-day subchronic oral toxicity studies of Silk peptide E5K6 were performed in Sprague–Dawley rats. In the acute toxicity study, Silk peptide E5K6 was administered orally to male and female rats at a single dose of 2000 and 5000mg/kg. Mortality, clinical signs and body weight changes were monitored for 14days. There were no treatment-related changes in these parameters. Therefore, the Approximate Lethal Dose (ALD) of Silk peptide E5K6 in male and female rats is higher than 5000mg/kg. In the subchronic toxicity study, Silk peptide E5K6 was administered orally to male and female rats for 90days at a single dose of 500, 1000, and 2000mg/kg. There were no toxicologically significant changes in clinical signs, body weight, food and water consumptions, ophthalmoscopic examination, urinalysis, hematological and serum biochemical examinations, necropsy findings, organ weights and histopathological examination of all of the animals treated with Silk peptide E5K6. These results suggest that the oral No Observed Adverse-Effect Level (NOAEL) of Silk peptide E5K6 is greater than 2000mg/kg/day in both sexes and the target organs were not established.

Peritoneal fluid changes in horses subjected to small colon distension

Intestinal devitalization in cases of small colon obstruction may be difficult to detect based only in clinical signs. The purpose was to serially evaluate blood and peritoneal fluid of horses subjected to small colon distension. Seventeen adult horses were allotted in three groups. In the small colon-distended group (DG, n=7) a surgically-implanted latex balloon was inflated to promote intraluminal small colon distension. In the shamoperated group (SG, n=5), the balloon was implanted but not inflated, and no surgery was done in the control group (CG, n=5). Blood and peritoneal fluid were sampled before and after (6 samples with a 30-minute interval) intestinal obstruction for cytological and biochemical analyses. No significant changes in clinical signs occurred within groups or across time during the experimental period. There were no statistical differences among SG and SG groups in hematologic and blood chemistry variables. Although total protein concentration and lactate dehydrogenase (LDH) activity in peritoneal fluid remained most of the time within reference values during the experimental period in all groups, increases from baseline values were detected in SG and DG groups. Such increases occurred earlier, progressively and with greater magnitude in the DG when compared with the SG (P<0.05). Increases from baselines values were also observed in total nucleated cells and neutrophils counts in the DG (P<0.05). In conclusion, distension of the equine small colon induced progressive subtle increases in total protein and LDH concentrations in the peritoneal fluid during the first hours. Serial evaluation of these variables in peritoneal fluid may be useful for early detection of intestinal devitalization in clinical cases of equine small colon obstruction.

Intensive short-term treatment with rituximab, cyclophosphamide and methylprednisolone pulses induces remission in severe cases of SLE with nephritis and avoids further immunosuppressive maintenance therapy

B cells play a central role in systemic lupus erythematosus (SLE). Rituximab is expected to induce apoptosis of all the CD20-positive B cells. A proportion of patients are refractory or intolerant to standard immunosuppression. These are candidate to new therapeutic options. Eight patients [six women, two men, mean age 41-year-old (27-51), with severe multiorgan involvement (kidney, skin, nervous system, polyarthritis, polyserositis, antiphospholipid antibody syndrome)] were considered eligible for an intensive combination therapy including rituximab. Rituximab was administered (dose 375 mg/m(2)) on Days #2, 8, 15 and 22. Two more doses were administered 1 and 2 months following the last weekly infusion. This treatment was combined with two pulses of 750 mg cyclophosphamide (Days #4 and 17) and three pulses of 15 mg/kg (Days #1, 4 and 8) methylprednisolone followed by oral prednisone, 50 mg for 2 weeks rapidly tapered until 5 mg in 2 months. Response was evaluated by assessing the changes in clinical signs and symptoms [Systemic Lupus Erythematosus Disease Activity Index (SLEDAI score)] and laboratory parameters for at least 12 months. Levels of erythrocyte sedimentation rate and anti-double-strand DNA antibodies significantly decreased (P < 0.01 at 12 months), whereas C3 and mainly C4 values increased at 6 months (P < 0.01 for C4). Proteinuria improved in the cases with renal involvement (P < 0.01 at 3, 6 and 12 months). SLEDAI score improved moving from the mean 17.3 (12-27) before therapy to 3.1 (1-5) after rituximab treatment. Constitutional symptoms including arthralgia, weakness and fever disappeared in all the previously affected patients; paresthesia improved in the four patients with polyneuropathy and skin lesions gradually resolved in the patients with necrotizing skin ulcers at presentation. Drug side effects were negligible. Long-lasting remissions were obtained in patients with severe SLE and major organ involvement by this intensive administration of rituximab combined with low doses of intravenous cyclophosphamide and methylprednisolone pulses followed by a rapid tapering of prednisone to 5 mg/day as a sole maintenance therapy.

Switching patients from preserved prostaglandin-analog monotherapy to preservative-free tafluprost

Purpose:Efficacy, tolerability and safety of the novel preservative-free prostaglandin tafluprost 0.0015% were investigated for the treatment of patients with glaucoma or ocular hypertension in a clinical setting.Patients and methods:Data were collected in a non-interventional, prospective, multi-center, observational, open label study. 118 patients were treated with a prostaglandin analog (PGA) monotherapy (preserved formulations of latanoprost, travoprost or bimatoprost) prior to baseline. Intraocular pressure (IOP) readings were recorded for each eye at baseline (previous therapy), 4–6 weeks, and 12 weeks after changing medical treatment to preservative-free tafluprost once-daily. We analyzed the change in IOP over the study period for all patients as well as for a subgroup of patients with prior PGA monotherapy. Subjective symptoms and objective ocular signs were determined. Comfort was measured using a 4 step scale. All adverse events were recorded. Paired t-tests were conducted to compare IOP values at baseline to IOP values after treatment with tafluprost 0.0015%. Bowker’s test of symmetry was used for statistical evaluation of changes of clinical signs (hyperemia).Results:In total 118 patients were eligible for evaluation. In these patients with prior PGA monotherapy (n = 118) IOP decreased significantly from 16.2 ± 4.3 mm Hg (95% CI: 0.55) at treated baseline to 14.8 ± 3.2 mm Hg (95% CI: 0.43; P < 0.001) at final visit on tafluprost. In a subset of patients with prior latanoprost monotherapy (n = 68) mean IOP at baseline (±SD) was reduced from 16.2 ± 4.6 mm Hg (95% CI: 0.77) 14.8 ± 3.1 mm Hg at final visit (95% CI: 0.54, P < 0.001), in patients with prior travoprost monotherapy (n = 32) from 16.2 ± 4.3 mm Hg (95% CI: 1.05) to 14.9 ± 3.3 mm Hg (95% CI: 0.91; P < 0.05) and in patients with prior bimatoprost monotherapy (n = 18) from 16.4 ± 3.5 mm Hg (95% CI: 1.14) to 15.0 ± 3.3 mm Hg (95% CI: 1.14; P = 0.252). Both, objective clinical signs and subjective symptoms improved after changing medication to preservative-free tafluprost until final visit. The number of patients with moderate and severe hyperemia decreased from 51 (43.2%) at baseline to 2 (1.9%) at final visit.Conclusion:Preservative-free tafluprost 0.0015% was effective, well tolerated and safe. IOP was controlled effectively and ocular symptoms and clinical signs were improved after changing medication to a monotherapy with preservative-free tafluprost in patients previously treated with a preserved latanoprost, travoprost or bimatoprost monotherapy.