The fecal bacterial microbiota is not useful for discriminating between lymphoplasmacytic enteritis and low-grade intestinal T-cell lymphoma in cats nor for predicting therapeutic response.

OBJECTIVE: To evaluate the fecal bacterial microbiota at the time of diagnosis (T0) and after 1 month of therapy (T1) in cats diagnosed with lymphoplasmacytic enteritis (LPE) or cats with low-grade intestinal T-cell lymphoma (LGITL) and to compare these findings with those of healthy cats. ANIMALS: 5 healthy cats, 13 cats with LPE, and 7 cats with LGITL were prospectively enrolled between June 2020 and June 2021. METHODS: Fecal samples were collected at T0 and T1, and DNA was extracted for 16S ribosomal amplicon sequencing. Alpha diversity and beta diversity were computed. The taxonomic assignment was performed using sequences from the Silva v138 formatted reference database. Differential abundant taxa were selected in each taxonomic level, with the P value adjusted < .05, as the cut-off. RESULTS: No significant differences in alpha and beta diversity were found either at T0 or T1 between healthy and diseased cats or between cats with LPE and LGITL. Beta-diversity analysis showed an increase in the Fusobacteriaceae family in cats with LGITL at T0, compared to cats with LPE. Regardless of histological diagnosis, several microbiota differences were found at T0 based on serum cobalamin levels. CLINICAL RELEVANCE: Fecal samples were successfully used to characterize the bacteriome of the intestinal tract in cats by 16S rRNA gene sequencing. However, results highlighted that the metagenomic evaluation was not useful to discriminate between LPE and LGITL nor to predict the therapeutic response in this study population.

A case series of urinary bladder rhabdomyosarcoma in seven dogs.

Background: Juvenile urinary bladder rhabdomyosarcoma (ubRMS) is a known entity; however, literature regarding its clinical behavior and endoscopic features is scarce. The aim of this study was to describe clinical and endoscopic features, and outcomes of ubRMS in dogs. Case Description: Dogs undergoing transurethral endoscopy and with a histological diagnosis of ubRMS were retrospectively collected. Seven dogs with a median age of 18 months (range 6-32 months) were included in this retrospective, multicenter, and descriptive study. Median tumor size was 58 mm (range 30-65 mm), and tumor location was bladder neck in three cases, trigone in two cases, and bladder body in two cases. Two dogs had monolateral ureteral obstruction. Two dogs presented with regional lymphadenopathy and one dog had lung lesions suggestive of metastatic disease. A grape-like mass was reported in four cases and solid in two, with variable consistency (two friables, two firms, and two not reported). Tumor treatments included surgery in three cases, surgery, and adjuvant doxorubicin in one case, and palliative therapy in three cases. The overall median survival time (ST) was 45 days. STs were shorter (range 20-45 days) for dogs treated with palliative care than for dogs treated with curative-intent treatment (range 70-120 days). Conclusion: ubRMS should be considered as a differential diagnosis in young dogs presenting with bladder masses. In this study, ubRMS confirmed its aggressive clinical behavior. Surgery and chemotherapy seem to increase STs but the prognosis remains poor.

Comparative Evaluation of Peripheral Blood Neutrophil to Lymphocyte Ratio, Serum Albumin to Globulin Ratio and Serum C-Reactive Protein to Albumin Ratio in Dogs with Inflammatory Protein-Losing Enteropathy and Healthy Dogs.

Few routinely available biomarkers are clinically useful in assessing dogs with inflammatory protein-losing enteropathy caused by immunosuppressive-responsive enteropathy (IRE-PLE). Only the neutrophil to lymphocyte ratio (NLR) has been studied, while no information exists on the use of the albumin to globulin ratio (AGR) and C-reactive protein to albumin ratio (CRP/ALB). We aimed to evaluate the clinical significance of the NLR, AGR and CRP/ALB in a population of dogs with IRE-PLE. The medical records of 53 IRE-PLE dogs were reviewed at the time of diagnosis (T0) and 1 month after the initiation of immunosuppressants (T1). A control group of 68 healthy dogs was used for comparison. At T0, the median values of the NLR and AGR of sick dogs were significantly higher and lower than those of healthy dogs, respectively. With the increase in the chronic enteropathy activity index, AGR and CRP/ALB significantly decreased and increased, respectively. At T1, NLR and AGR significantly increased, while CRP/ALB significantly decreased. NLR, AGR and CRP/ALB did not differ significantly between dogs classified as responders and nonresponders according to the chronic enteropathy activity index. Further studies are needed to provide more information on this subject.

Relationship between Serum Protein Electrophoresis, Endoscopic and Histopathological Scores in 99 Cats with Chronic Enteropathy.

Few studies have investigated total protein (TP) and serum protein electrophoresis (SPE) in cats with chronic enteropathy (CE). Cats diagnosed with CE were evaluated to investigate the relationships between TP, SPE and endoscopy, histopathology, and extraintestinal involvement. Medical records were searched for cats with a history of chronic gastrointestinal signs and a final diagnosis of CE. Information on signalment, TP, SPE, endoscopic score, histopathological diagnosis and score, and concurrent hepatic or pancreatic ultrasonographic alterations was collected. Relationships between protein profiles and other variables were investigated. Ninety-nine cats were included in the study, 63 diagnosed with various degrees of bowel inflammation and 36 with small-cell alimentary lymphoma. The most common TP alteration was hypoproteinemia (24%). No significant differences were observed between protein profiles and endoscopic and histopathological severity scores. Forty-five cats showing concurrent pancreatic and/or hepatic ultrasonographic alterations, had significantly lower albumin, lower α-globulin, and higher γ-globulin levels than cats not showing concurrent alterations. Disease severity scores did not seem to influence the protein profile in cats with CE. Extraintestinal involvement may be suspected in cats with lower albumin and α-globulins and higher γ-globulins.

Gastroduodenal ulceration detected endoscopically in cats: retrospective study of 61 patients.

OBJECTIVES: The aim of this study was to describe the endoscopic appearance of gastroduodenal ulcers (GDUs), and to assess the clinical, ultrasonographic and histological data, as well as long-term follow-up, in cats. METHODS: The medical record databases of five veterinary endoscopists were evaluated between January 2016 and 2020, in a retrospective study. Cats with at least one gastric or duodenal ulcer detected by endoscopic examination were included. All the medical records of the selected cats were reviewed and information was collected regarding breed, age, sex, neuter status, medical history, clinical signs, and ultrasonographic, endoscopic and histological findings. The cats were evaluated at 6, 12 and 18 months. RESULTS: Sixty-one cats with a median age of 9.0 years (range 2.0-16.0) were included in the study. The most common complaints were vomiting (n = 55; 90%) and hyporexia (n = 40; 66%); haematemesis was reported in 12 (20%) cats. Endoscopy showed GDUs in the following locations: gastric body in 28 cats (46%), antropyloric area in 34 cats (56%), fundus in 13 cats (21%) and duodenum in eight cats (13%). A single GDU was found in 42 cats (69%) and multiple GDUs were seen in 19 cats (31%). Histopathological evaluation revealed benign lesions in 33 (54%) cats and malignant lesions in 28 (46%; 24 high-grade lymphoma, one low-grade lymphoma and three carcinoma). High-grade lymphoma was detected only in the stomach. Cats diagnosed with malignant GDUs (median 10.5, range 4-16) were significantly older than cats with benign lesions (P = 0.002). CONCLUSIONS AND RELEVANCE: GDUs are common and were detected in 5.1% of cats undergoing an upper gastrointestinal endoscopy. The risk of a malignant ulcer increases proportionally with each year of increasing age. GDU location, number and morphological appearance do not provide any indication of the nature of the ulcer; however, duodenal ulcers are frequently benign. Endoscopic examination facilitates the early and minimally invasive detection of GDUs in cats.

Usefulness of squash preparation cytology in the diagnosis of canine urinary bladder carcinomas.

BACKGROUND: Epithelial cells show varying degrees of cytologic atypia in dogs with nonmalignant lesions (NML) and carcinomas (ubC) of the bladder, making histopathologic examination necessary for a definitive diagnosis. OBJECTIVES: This study aimed to investigate the diagnostic performance of squash preparation cytology and identify several cytomorphologic features of ubC to assist in diagnoses. METHODS: Squash preparations were made and reviewed in dogs that underwent transurethral cystoscopy. The results were compared with histopathologic diagnoses. Two cytopathologists performed blinded assessments using a scoring system established for 11 cytologic features, including the presence of macronuclei, abnormal nucleoli, atypical mitoses, signet ring cells, multinucleated cells, nuclear molding, anisokaryosis, cytoplasmatic microvacuolization, cell arrangements, and neutrophil and lymphocyte infiltrations. Based on cytologic and histopathologic diagnoses, dogs were divided into ubC and NML groups. Associations between cytologic and histopathologic diagnoses were investigated, and agreement between the cytopathologists was calculated. Cytologic features were analyzed with multivariate logistic regression models. The performance of predictors in the final model was evaluated in terms of Sensitivity (Se), Specificity (Sp), accuracy, positive predictive value (PPV), negative predictive value (NPV), likelihood ratio positive (LR+), and negative (LR-) values, and the diagnostic odds ratio (DOR). RESULTS: Forty-four dogs diagnosed with ubC, and 17 with NML were included in the study. Cytologic and histopathologic diagnoses were significantly associated with each cytopathologist. There was an almost perfect agreement between cytopathologists (κ = 0.88). The absence of neutrophilic infiltration, the presence of multinucleated cells, and nuclear molding were associated with ubC; using a combination of these features in parallel testing resulted in Se = 0.98, Sp = 0.65, accuracy = 0.89, PPV = 0.88, NPV = 0.92, LR + =2.77, LR- = 0.04, and DOR = 7.7. CONCLUSIONS: Squash preparation cytology could be a reliable technique to diagnose ubC in dogs. The best diagnostic combination was the absence of neutrophilic infiltration, multinucleated cells, and nuclear molding.

Clinical evaluation and microbiota analysis in 9 dogs with antibiotic-responsive enteropathy: A prospective comparison study.

BACKGROUND: Antibiotic-responsive enteropathy (ARE) is diagnosed by excluding other causes of diarrhea and when there is a short-term response to administration of antibiotics. OBJECTIVES: To characterize the gut microbiota and clinical trend of dogs with suspected ARE and to evaluate the variation in microbiota before (T0), after 30 days (T30) of tylosin treatment, and 30 days after discontinuation of treatment (T60). A further objective was to evaluate whether changes in gut microbiota are related to relapses of diarrhea when the therapy is tapered. ANIMALS: Study sample (group A) was composed of 15 dogs with chronic diarrhea, group B was composed of 15 healthy dogs. Group A was given tylosin for 30 days. METHODS: A multicentric prospective study. Clinical Indexes, fecal score, and samples for microbiota analysis were collected at T0, T30, and T60 in group A and T0 and T30 in group B. The gut microbiota was analyzed via 16S ribosomal RNA gene. Qiime2 version 2020.2 was used to perform bioinformatic analyses, and Alpha- and Beta-diversity were computed. RESULTS: Diarrhea recurred after T30 in 9 of 14 dogs, which were classified as affected by ARE. At T0, a difference was noted in the beta-diversity between groups (Bray Curtis metric P = .006). A T0-T30 difference in alpha-diversity was noted in group A (Shannon index P = .001, Faith PD P = .007). CONCLUSIONS AND CLINICAL IMPORTANCE: Although tylosin influences the microbiota of dogs with ARE, we failed to find any specific characteristic in the microbiota of dogs with ARE.

Presence of bronchial nodules, younger age, and heavier body weight are associated with a diagnosis of eosinophilic lung disease in dogs with cough.

OBJECTIVE: To describe the association between a diagnosis of eosinophilic lung disease (ELD) in dogs with signalment and bronchoscopic features and evaluate the accuracy of visualization of nodules for the diagnosis of ELD. ANIMALS: 781 dogs with cough that underwent bronchoscopy between 2014 and 2016. PROCEDURES: Data were extracted from the medical records of each included dog. Multivariable logistic regression was performed to investigate associations between ELD and patient characteristics. RESULTS: ELD was diagnosed in 113 (14.5%) dogs. More than 3 nodular lesions of the bronchial mucosa were detected in 64 (8.2%) dogs. The odds of having ELD were greater in dogs with nodules (adjusted OR [aOR], 26.0; 95% CI, 13.0 to 52.0) and static bronchial collapse (aOR, 2.3; 95% CI, 1.1 to 4.6), and lower in dogs having focal versus diffuse inflammation (aOR, 0.05; 95% CI, 0.01 to 0.37). The odds of having ELD decreased for each 1-year increase in age (aOR, 0.86; 95% CI, 0.80 to 0.92), and increased for each 1-kg increase in weight (aOR, 1.04; 95% CI, 1.01 to 1.06). Visualization of nodules during bronchoscopy had a overall accuracy of 89.4% (95% CI, 87.0% to 91.4%), sensitivity of 41.6% (32.4% to 51.2%), and specificity of 97.5% (96.0% to 98.5%) for a diagnosis of ELD. CLINICAL RELEVANCE: On the basis of high specificity and negative predictive value, lack of visualization of bronchial nodules during bronchoscopy can be used to preliminarily rule out ELD. However, visualization of bronchial nodules does not imply presence of ELD. This could be especially relevant when results of BAL cytology are available several days after the actual bronchoscopy.

Detection of Anti-Erythrocyte Antibodies in Dogs with Inflammatory Bowel Disease (IBD).

Several extra-intestinal manifestations, including immune-mediated cytopenias, are reported in human inflammatory bowel disease (IBD), whereas they are poorly documented in dogs. Hypothesizing that immune-mediated subclinical anemia can occur in canine IBD, the study aim was to evaluate the erythrogram and the presence of anti-RBC antibodies in dogs with IBD. IBD was diagnosed according to the following criteria: chronic gastrointestinal signs, ruling out of extra-intestinal diseases, no improvement with diet trial, histological evidence of inflammatory infiltration, and improvement after immunosuppressant therapy. Canine Chronic Enteropathy Clinical Activity Index (CCECAI) endoscopic and histopathological scores were assessed for each dog. Twenty-five dogs were enrolled, and each dog had a CBC evaluation prior to endoscopy. The CBC was performed using laser hematology analyzer and blood smears were carefully reviewed for the presence of nucleated RBC, anisocytosis, polychromasia, and Howell-Jolly bodies. IgG and IgM anti-RBC antibodies were evaluated with flow cytometry. A high frequency of positive cases for anti-RBC antibodies in dogs with IBD (17/25 dogs) was ascertained. Approximatively 50% of dogs showed some hematologic features of RBC regeneration in addition to hematologic findings consistent with chronic inflammation. Anti-RBC antibodies and signs of erythroid regeneration may suggest possible subclinical chronic immune-mediated hemolysis that can cause anemia in dogs with IBD, together with the chronic inflammation.

Immunosuppressant-Responsive Enteropathy and Non-Responsive Enteropathy in Dogs: Prognostic Factors, Short- and Long-Term Follow Up.

A multicentre prospective study was performed to assess whether clinical, hematobiochemical, endoscopic and histopathological parameters were associated with mortality, clinical response and relapse of disease in short- and long-term follow-up of a total of 165 dogs with chronic inflammatory enteropathy, of which 150 had immunosuppressant responsive enteropathy (IRE), and 15 had non-responsive enteropathy (NRE) dogs. Clinical severity (CCECAI) was evaluated from presentation (T0) to 18 months (T18) from diagnosis. T0 body condition score (BCS), selected haematological parameters and endoscopic and histopathological scores were evaluated. Presence/absence of histopathological duodenal lesions was recorded. Responders were evaluated using CCECAI at T1. Relapse was evaluated from T3 to T18. Long-term responders included dogs who responded at T1 and showed no relapse. Dogs were divided into responders/non-responders, survivors/non-survivors and relapsed/non-relapsed. At T1, 15/165 dogs (9%) were considered NRE. Sixteen dogs (11%) were considered relapsed at T3, 8% at T6 and 10% at T12, and none of 96 dogs relapsed at T18. NREs showed significantly lower BCS than IREs. Non-survivors showed a significantly lower serum albumin concentration and BCS than survivors. Non-responders, relapsed or non-survivors had higher presence of lacteal dilatation compared to long-term responders. Dogs with IRE showed a good clinical course with a low relapse rate, with only a few dogs in the NRE group. Reduction of BCS, albumin and lacteal dilatation at diagnosis may be considered negative prognostic factors for response, mortality and long-term disease remission.

Evaluation of duodenal endoscopic and histologic findings, including counts of forkhead box P3-positive regulatory T cells, in dogs with immunosuppressant-responsive enteropathy.

OBJECTIVE: To prospectively evaluate the clinical and prognostic importance of duodenal endoscopic and histologic findings, including duodenal mucosal counts of forkhead box P3-positive regulatory T cells (Foxp3+ Tregs), in dogs with immunosuppressant-responsive enteropathy (IRE). ANIMALS: 57 client-owned dogs with IRE. PROCEDURES: The canine chronic enteropathy clinical activity index (CCECAI) was used to assess each dog when IRE was diagnosed (T0) and 1, 3, 6, and 12 months later. Dogs were grouped on the basis of clinical response (responder group vs nonresponder group) and 12-month long-term outcome (responded to treatment and did not relapse [good outcome group] vs did not respond to treatment or had relapsed [bad outcome group]). At T0, dogs underwent gastrointestinal endoscopy and endoscopic biopsy, with results for variables of duodenal endoscopic and histologic evaluations scored and compared across groups. RESULTS: At T0, the overall median CCECAI score was 7; CCECAI score was not associated with clinical response or relapse. Dogs had significantly greater odds of being in the bad outcome group (vs the good outcome group) if they had a histologic score of 3 (OR, 3.5; 95% CI, 1.09 to 11.3). No differences in the counts of Foxp3+ Tregs were detected between groups. CONCLUSIONS AND CLINICAL RELEVANCE: In dogs with IRE, results indicated that evaluation of Foxp3+ Tregs did not have prognostic value, whereas a duodenal histologic score of 3 could be a negative prognostic factor for response and relapse, and higher severity scores for intraepithelial lymphocytes and lamina propria lymphocytes and plasma cells in duodenal biopsy samples may be negatively associated with response.

Platelet abnormalities and platelet-to-lymphocyte ratios in canine immunosuppressant-responsive and non-responsive enteropathy: A retrospective study in 41 dogs.

Few studies have examined platelet alterations in dogs with chronic enteropathy. Our aim was to investigate platelet count (PLT), mean platelet volume (MPV), and platelet-to-lymphocyte ratio (PLR) in dogs diagnosed with immunosuppressant-responsive enteropathy (IRE). In this retrospective study of 41 dogs, data regarding signalment, canine chronic enteropathy clinical activity index (CCECAI), endoscopic and histopathological scores, PLT, MPV, PLR, total serum protein concentrations, albumin, and iron were collected. Clinical response and relapse were assessed with the evaluation of CCECAI over time. One month after starting therapy, dogs with >25% CCECAI reduction were considered responders. During a three-month CCECAI evaluation as part of a twelve-month follow-up, a CCECAI >3 together with a ≥2 unit increase in responder dogs was considered a relapse. PLT and PLR displayed significant negative correlation with MPV. MPV was positively correlated with total protein and albumin levels and negatively correlated with CCECAI. Three dogs were classified as non-responders, and 14 relapsed within 12 months. No differences were observed in PLT, MPV, or PLR between responding/non-responding and relapsing/non-relapsing groups. PLT, MPV, and PLR correlated with total protein, albumin, and CCECAI, confirming PLT as a potential marker, and suggesting MPV as a new marker of clinical efficacy against canine IRE.

Neutrophil-to-Lymphocyte Ratio (NLR) in Canine Inflammatory Bowel Disease (IBD).

Inflammatory bowel disease (IBD) is a multifactorial chronic inflammatory disorder leading to structural changes in the intestinal wall. In humans, the neutrophil-to-lymphocyte ratio (NLR) has been proposed as a promising marker of IBD. This study evaluated the possible clinical and prognostic significance of the NLR in dogs with IBD. This retrospective study enrolled 41 dogs diagnosed with IBD presented to University of Pisa from January 2017 to January 2018. For each dog, age, sex, canine chronic enteropathy clinical activity index (CCECAI), endoscopic and histopathological grading were recorded. Complete blood count, serum total protein, albumin, cholesterol, and C-reactive protein at the time of endoscopy were recorded. A control group (CG) of healthy dogs from a blood donor database was built. NLR was calculated for both IBD and CG as the ratio between absolute neutrophils and lymphocytes. Presence of crypt distension, lacteal dilation (LD), mucosal fibrosis, intraepithelial lymphocytes was recorded. Follow-up information was obtained from electronic medical records and dogs were classified as responders and non-responders based on CCECAI variation between admission and the first recheck. IRE dogs showed higher NLR compared to healthy dogs. NLR correlated negatively with total protein, albumin, and cholesterol and correlated positively with CCECAI. Dogs with LD showed higher NLR than dogs without LD. Non-responders showed higher NLR compared to responders. In conclusion, as in IBD human patients, the NLR acts as an inflammatory marker providing further information on severity of the disease and could be useful in predicting treatment response.

Serum homocysteine concentration in dogs with immunosuppressant-responsive enteropathy.

BACKGROUND: Homocysteine (HCY) was evaluated in healthy and chronic enteropathic dogs, however no studies on dogs with immunosuppressant-responsive enteropathy are available. OBJECTIVES: The aim was to evaluate serum HCY concentrations and its prognostic role in dogs with immunosuppressant-responsive enteropathy compared to healthy dogs. METHODS: Serum HCY concentration was statistically compared between 24 healthy dogs and 29 dogs with immunosuppressant-responsive enteropathy. Correlation analyses between serum total protein, albumin (ALB), C-reactive protein (CRP), folate and cobalamin, and serum HCY concentration were performed in immunosuppressant-responsive enteropathic dogs. RESULTS: The associations between serum HCY concentration and clinical, histological, endoscopic scores and follow-up were evaluated. Mean serum HCY concentration was higher in immunosuppressant-responsive enteropathic dogs compared to control dogs (30.22 ± 8.67 µmol/L vs. 5.26 ± 2.78 µmol/L; p < 0.0001). No association between serum HCY concentration and total protein, ALB, CRP, folate concentration as well as, clinical score, histological and endoscopic scores was found. A negative correlation between serum HCY concentration and cobalamin was noted (p = 0.0025, r = -0.54). No significant difference in HCY was found between responsive and non-responsive dogs or between survivors and non-survivors. CONCLUSIONS: Although, serum HCY concentration was higher in immunosuppressant-responsive enteropathy, its prognostic value remains unclear. However, further prospective, large-scale studies are warranted to better investigate the possible prognostic role of HCY in immunosuppressant-responsive enteropathic dogs.

Serum amino acid profile in 51 dogs with immunosuppressant-responsive enteropathy (IRE): a pilot study on clinical aspects and outcomes.

BACKGROUND: Lower levels of tryptophan (TRP) have been identified in people with inflammatory bowel disease and in dogs with protein-losing enteropathy (PLE). No data on serum amino acids (AAs) but some on plasma in canine immunosuppressant-responsive enteropathy (IRE) are available. The aim of this study is to compare serum AAs between healthy and IRE dogs, considering clinicopathological variables and follow-up. RESULTS: Twenty-six healthy control dogs (CD) and 51 IRE dogs were included. IRE was diagnosed after the exclusion of extra-intestinal diseases and food and antibiotic responsive enteropathies. The canine chronic enteropathy clinical activity index (CCECAI) was assessed at presentation and during the clinical follow-up. In CD and IRE dogs, 19 different serum AAs were measured. IRE dogs were classified into responders, partial responders and non-responders, based on CCECAI after 1 month, and divided into PLE and non-PLE, based on albumin level. IRE dogs showed lower L-Tyrosine (TYR), L-Phenylalanine (PHE) and TRP (p < 0.001) and higher L-Serine (SER), L-Glutamic acid (GLU), L-Arginine (p < 0.001), L-Threonine (p = 0.013), Proline (p = 0.044), L-Cysteine (p = 0.003), L-Valine (p = 0.018), L-Lysine (p = 0.01) and L-Isoleucine (p = 0.005) than CDs. PLE dogs showed lower L-Histidine (HIS) (p = 0.008), PHE (p = 0.005) and TRP (p = 0.005) than non-PLE dogs. In IRE dogs, median GLU was significantly lower in dogs with BCS 3/9 than BCS 5/9 category (p = 0.036). Total protein was positively correlated with PHE and TRP (both p = 0.031, r = 0.30) and albumin was positively correlated with HIS (p = 0.025, r = 0.31), PHE and TRP (both p = 0.001, r = 0.46). HIS (p = 0.041), PHE (p = 0.047) and TRP (p = 0.044) concentrations were significantly lower in non-responders than in responders and partial responders. CONCLUSIONS: This study may suggest further investigation on serum, HIS, PHE, TRP and TYR as markers of intestinal disease and proposed HIS, PHE and TRP as prognostic marker for response to therapy.