Effect of empagliflozin on cardiac remodelling in South Asian and non-South Asian individuals: insights from the EMPA-HEART CardioLink-6 randomised clinical trial.

BACKGROUND: This exploratory sub-analysis of the EMPA-HEART CardioLink-6 trial examined whether the previously reported benefit of the sodium-glucose cotransporter 2 (SGLT2) inhibitor empagliflozin on left ventricular (LV) mass (LVM) regression differs between individuals of South Asian and non-South Asian ethnicity. METHODS: EMPA-HEART CardioLink-6 was a double-blind, placebo-controlled clinical trial that randomised 97 individuals with type 2 diabetes mellitus (T2DM) and coronary artery disease (CAD) to either empagliflozin 10 mg daily or placebo for 6 months. LV parameters and function were assessed using cardiac magnetic resonance imaging. The 6-month changes in LVM and LV volumes, all indexed to baseline body surface area, for South Asian participants were compared to those for non-South Asian individuals. RESULTS: Compared to the non-South Asian group, the South Asian sub-cohort comprised more males, was younger and had a lower median body mass index. The adjusted difference for LVMi change over 6 months was -4.3 g/m2 (95% confidence interval [CI], -7.5, -1.0; P = 0.042) for the South Asian group and -2.3 g/m2 (95% CI, -6.4, 1.9; P = 0.28) for the non-South Asian group (Pinteraction = 0.45). There was no between-group difference for the adjusted differences in baseline body surface area-indexed LV volumes and LV ejection fraction. CONCLUSIONS: There was no meaningful difference in empagliflozin-associated LVM regression between South Asian and non-South Asian individuals living with T2DM and CAD in the EMPA-HEART CardioLink-6 trial. TRIAL REGISTRATION: ClinicalTrials.gov Identifier: NCT02998970 (First posted on 21/12/ 2016).

Comparative evaluation of intraoperative aberrometry and Barrett's toric calculator in toric intraocular lens implantation.

Purpose: Barrett toric calculator (BTC) is known for its accuracy in toric IOL (tIOL) calculation over standard calculators; however, there is no study in literature to compare it with real-time intraoperative aberrometry (IA). The aim was to compare the accuracy of BTC and IA in predicting refractive outcomes in tIOL implantation. Methods: This was an institution-based prospective, observational study. Patients undergoing routine phacoemulsification with tIOL implantation were enrolled. Biometry was obtained from Lenstar-LS 900 and IOL power calculated using online BTC; however, IOL was implanted as per IA (Optiwave Refractive Analysis, ORA, Alcon) recommendation. Postoperative refractive astigmatism (RA) and spherical equivalent (SE) were recorded at one month, and respective prediction errors (PEs) were calculated using predicted refractive outcomes for both methods. The primary outcome measure was a comparison between mean PE with IA and BTC, and secondary outcome measures were uncorrected distance visual acuity (UCDVA), postoperative RA, and SE at one month. SPSS Version-21 was used; P < 0.05 considered significant. Results: Thirty eyes of 29 patients were included. Mean arithmetic and mean absolute PEs for RA were comparable between BTC (-0.70 ± 0.35D; 0.70 ± 0.34D) and IA (0.77 ± 0.32D; 0.80 ± 0.39D) (P = 0.09 and 0.09, respectively). Mean arithmetic PE for residual SE was significantly lower for BTC (-0.14 ± 0.32D) than IA (0.001 ± 0.33D) (-0.14 ± 0.32D; P = 0.002); however, there was no difference between respective mean absolute PEs (0.27 ± 0.21 D; 0.27 ± 0.18; P = 0.80). At one-month, mean UCDVA, RA, and SE were 0.09 ± 0.10D, -0.57 ± 0.26D, and -0.18 ± 0.27D, respectively. Conclusion: Both IA and BTC give reliable and comparable refractive results for tIOL implantation.

Cyclin-dependent kinase inhibitor p21 and proliferative marker ki67 in colonic carcinoma.

Objective: Tumor grade employed for colorectal cancer has long been based on the degree of differentiation, which is difficult to judge objectively. The aim of this study was to assess the immunohistochemical expression of p21 and ki67 and their correlation with the histological grading of colorectal carcinoma. Materials and Methods: A total of 45 biopsy specimens of colorectal cancer were pathologically reviewed and correlation of grade and differentiation of tumor was performed with immunostaining. Results: Ki 67 and p21 markers showed inverse relationship. An inverse relationship of p21 was found with tumor grade, differentiation, Dukes staging and lymph node status, whereas no correlation could be found between these parameters and ki67 expression. Conclusion: We found that p21 can be used to assess the grading and metastatic potential of colorectal carcinoma whereas increased Ki67 expression can help us in the diagnosis of malignancy.

Characteristics of an Acidic Phytase from Aspergillus aculeatus APF1 for Dephytinization of Biofortified Wheat Genotypes.

Phytases are the special class of enzymes which have excellent application potential for enhancing the quality of food by decreasing its inherent anti-nutrient components. In current study, a protease-resistant, acidic phytase from Aspergillus aculeatus APF1 was partially purified by ammonium sulfate fractionation followed by chromatography techniques. The molecular weight of partially purified phytase was in range of 25-35 kDa. The purified APF1 phytase was biochemically characterized and found catalytically active at pH 3.0 and 50 °C. The Km and Vmax values of APF1 phytase for calcium phytate were 3.21 mM and 3.78 U/mg protein, respectively. Variable activity was observed with metal ions and among inhibitors, chaotropic agents and organic solvents; phenyl glyoxal, potassium iodide, and butanol inhibited enzyme activity, respectively, while the enzyme activity was not majorly influenced by EDTA, urea, ethanol, and hexane. APF1 phytase treatment was found effective in dephytinization of flour biofortified wheat genotypes. Maximum decrease in phytic acid content was noticed in genotype MB-16-1-4 (89.98%) followed by PRH3-30-3 (82.32%) and PRH3-43-1 (81.47%). Overall, the study revealed that phytase from Aspergillus aculeatus APF1 could be effectively used in food and feed processing industry for enhancing nutritional value of food.

Effect of Lens Fragmentation Patterns on Phacoemulsification Parameters and Postoperative Inflammation in Femtosecond Laser-Assisted Cataract Surgery.

PURPOSE: To evaluate intraoperative and postoperative outcomes with "chop" or "matrix" lens fragmentation patterns in femtosecond laser-assisted cataract surgery. METHODS: Prospective comparative study of 66 eyes with grade III-IV nuclear sclerosis was conducted at an apex tertiary care ophthalmic center. Cases were randomly allocated to undergo femtosecond laser pretreatment using matrix pattern (group I; n = 33) or chop pattern (group II; n = 33) of lens fragmentation (LenSx laser platform), followed by phacoemulsification. The primary outcome measures were intraoperative phacoemulsification parameters and postoperative anterior chamber (AC) flare. Secondary outcome measures were intraoperative complications, postoperative central macular thickness, visual acuity, and endothelial cell counts. Follow-up was performed on postoperative day (POD) 1 and 30. RESULTS: Phacoemulsification parameters including cumulative dissipated energy (p = 0.008), ultrasonic time (p = 0.001), aspiration time (p < 0.001), and total duration (p = 0.001) were significantly less in group I. The AC flare was 9.8 ± 4.6 in group I and 15.4 ± 6.0 in group II (p < 0.001) on POD 1, and the difference persisted at 1 month. A highly significant positive correlation was observed between the total duration of phacoemulsification and AC flare (p < 0.001). No case developed cystoid macular edema. The postoperative specular counts were significantly less in group II on POD 1 (p = 0.036) and POD 30 (p = 0.02). There was no difference in visual acuity between the two groups, and intraoperative complications were not observed in any case. CONCLUSION: A decrease in phacoemulsification time and energy is observed after femtosecond laser pretreatment with the matrix pattern of lens fragmentation as compared to the chop pattern. The endothelial cell loss and postoperative inflammation is significantly less with the matrix pattern, and the inflammation correlates with the duration of phacoemulsification.

In Vivo Confocal Microscopic Characteristics of Crystalline Keratopathy in Patients with Sclerokeratitis.

PURPOSE: To report in vivo confocal features in a clinical case series of patients with sclerokeratitis presenting as crystalline keratopathy. METHODS: Five cases of crystalline keratopathy following sclerokeratitis are described. Confocal microscopic images of the cornea were captured in all cases to confirm the diagnosis by evaluating the morphology of the crystals. RESULTS: Unilateral and non-progressive peripheral crystalline keratopathy manifested after previous episodes of sclerokeratitis in the involved eye. Confocal microscopy revealed numerous, discrete, hyperreflective, needle-like, shiny crystals in the anterior and posterior stromal layers of the cornea. These deposits were oriented randomly and showed occasional confluence. An extensive ophthalmic and systemic evaluation did not reveal any other contributory factors. CONCLUSION: Crystalline keratopathy, probably resulting from an immune-mediated response, is a possible manifestation of sclerokeratitis. This should be considered during long-term follow-up of such patients and differentiated from infectious crystalline keratopathy.

Long-Term Evaluation of Specular Microscopic Changes Following Nd: YAG Iridotomy in Chronic Primary Angle-Closure Glaucoma Eyes.

AIM: The aim of this study was to evaluate specular microscopy of chronic primary angle-closure glaucoma (CPACG) eyes at least 1 year after Nd:YAG iridotomy, and compare them with CPACG eyes without an iridotomy and age-matched, normal eyes. PATIENTS AND METHOD: Consecutive patients of CPACG at the Glaucoma service were screened. All patients underwent slit-lamp biomicroscopy, +90 D examination, and applanation tonometry. A total of 100 eyes of 100 consecutive patients of CPACG with an Nd:YAG iridotomy performed ≥1 year before, who met all inclusion/exclusion criteria, 60 consecutive CPACG eyes without an iridotomy, and 60 age and refraction-matched control eyes were enrolled. A specular microscopy was performed in one eye of each patient by an observer masked to diagnosis. RESULTS: CPACG patients had a mean age of 62±8 years, a mean intraocular pressure of 18±5.3 mm Hg, a mean specular count of 2536±224 cells/mm, and mean duration after iridotomy of 3.2±2 years. There was a significant correlation of specular endothelial counts with age (r=-0.39; P<0.001) and interval after iridotomy (r=-0.25; P=0.01). CPACG eyes without an iridotomy had a mean age of 62±5 years and a mean specular count of 2469±199 cells/mm. Normal control eyes with a mean age of 61±6 years had a mean specular count of 2729±299 cells/mm. There was no significant difference in specular count between CPACG eyes with or without an iridotomy (P=0.19); however, both CPACG groups had a specular count significantly lower than controls (P=0.01 and 0.02, respectively). There was no statistically significant difference seen in polymegathism (coefficient of variation) and pleomorphism (% of hexagonal cells) in endothelial cells among the 3 groups. CONCLUSIONS: An Nd:YAG iridotomy in CPACG eyes did not lead to any significant changes in central corneal specular microscopy in the long term as compared with patients who did not undergo iridotomy. Eyes with CPACG, without and after an iridotomy, had a lower specular count compared with age-matched controls.

Effect of reduced glutathione supplementation in semen extender on tyrosine phosphorylation and apoptosis like changes in frozen thawed Hariana bull spermatozoa.

To provide new insights into the mechanisms through which reduced glutathione (GSH) is able to protect spermatozoa, we tested the hypothesis that cryocapacitation and apoptosis like changes can contribute to the negative effect of freezing and thawing on bull spermatozoa, and that GSH prevent this damage. Having known protective effects of GSH in terms of a potent antioxidant, we evaluated capacitation, tyrosine phosphorylation and apoptosis like changes in bull spermatozoa after freezing and thawing in egg yolk tris glycerol extender containing (0.5m M-GSH-T1 & 1mM GSH-T2) and without GSH serving as the control (C). Forty ejaculates were collected from four Hariana bulls and were pooled due to non significant variations among the bull ejaculates for the evaluation of sperm attributes. Capacitation like changes, tyrosine phosphorylation, localization of tyrosine phosphorylated proteins, apoptosis like changes in terms of mitochondrial transmembrane potential and DNA fragmentation after final dilution, 4h of equilibration at 4°C and 24h after freezing and thawing were evaluated. GSH supplementation at 0.5mM showed significant reduction in B- and AR- pattern spermatozoa during all stages of semen freezing and thawing. Immunoblot revealed six proteins which were tyrosine phosphorylated and protein of 30 and 75kDa (p30, p75) were the major tyrosine phosphorylted proteins. On further analysis, the p30 showed differential variation in intensity in all the three groups after freezing and thawing. Positive immune reactivity for tyrosine phosphorylated proteins was found in neck, middle piece and post-acrosomal regions of spermatozoa. Addition of 0.5mM GSH decreased percentage of spermatozoa showing fragmented DNA and increased the percentage of spermatozoa having high transmembrane mitochondrial potential (P<0.05). This study demonstrates that GSH favours survival of bull spermatozoa by interfering with apoptotic and cryocapacitation pathways, and thereby protects the spermatozoa from deleterious effects of cryopreservation. The findings of the study indicated that GSH at 0.5mM can be effectively used as an additive in bull semen extender for freezing and thawing.

Corneal changes in childhood glaucoma.

OBJECTIVE: To study the clinical features and topography of the cornea in eyes with childhood glaucoma. DESIGN: Cross-sectional, observational study. PARTICIPANTS: Fifty-eight eyes with childhood glaucoma and 28 eyes of age-matched controls. METHODS: Clinical and topographic corneal changes were evaluated. MAIN OUTCOME MEASURES: Corneal topographic changes were evaluated on Orbscan (Orbscan Topography System II; Bausch & Lomb, Salt Lake City, UT) in eyes with childhood glaucoma and those changes were compared with the control eyes. RESULTS: Fifty-eight eyes with childhood glaucoma and 28 eyes of age-matched controls were evaluated. Thirty-six eyes (62.1%) were classified as having primary childhood glaucoma and 22 eyes (37.94%) as having childhood glaucoma with associated ocular anomalies. The corneas in 18 of 58 eyes (31.0%) with childhood glaucoma were clear, whereas 24.1% of eyes (14/58 eyes) had some corneal opacification. Haab's striae were noted in 44.8% of eyes (26/58 eyes) and were most frequently present between 3 and 5 mm from the optical axis. The mean posterior elevation recorded in eyes with childhood glaucoma controlled with medication or surgery was significantly higher than that in control eyes: 0.043 ± 0.027, 0.042 ± 0.017, and 0.018 ± 0.058 µm, respectively (P < 0.0001). The presence of Haab's striae was correlated significantly with a higher posterior elevation (P = 0.0396) and poor vision. The mean anterior elevation in eyes with childhood glaucoma (0.022 ± 0.015 µm) and in control eyes (0.015 ± 0.078 µm) was comparable (P = 0.08). Corneal astigmatism in eyes with childhood glaucoma was significantly higher and irregular compared with that in control eyes: 2.09 ± 1.40 versus 0.93 ± 0.60 diopter cylinder (P = 0.0001); the irregularity index was 2.8 (range, 1-18.1) and 2.3 (range, 0.6-2.3) at 3 mm (P = 0.0005) and 3.2 (range, 1.4-21.3) and 1.8 (range, 0.5-2.9) at 5 mm, respectively (P = 0.0003). Best-corrected visual acuity correlated significantly with cup-to-disc ratio, axial length, refractive error, astigmatism, and posterior corneal elevation. Multivariate analysis showed a significant correlation only with cup-to-disc ratio and axial length. CONCLUSIONS: Childhood glaucoma causes a significant increase in posterior corneal elevation and irregular astigmatism, which contribute to visual disability in such eyes.

Deletion of specific immune-modulatory genes from modified vaccinia virus Ankara-based HIV vaccines engenders improved immunogenicity in rhesus macaques.

Modified vaccinia virus Ankara (MVA) is a safe, attenuated orthopoxvirus that is being developed as a vaccine vector but has demonstrated limited immunogenicity in several early-phase clinical trials. Our objective was to rationally improve the immunogenicity of MVA-based HIV/AIDS vaccines via the targeted deletion of specific poxvirus immune-modulatory genes. Vaccines expressing codon-optimized HIV subtype C consensus Env and Gag antigens were generated from MVA vector backbones that (i) harbor simultaneous deletions of four viral immune-modulatory genes, encoding an interleukin-18 (IL-18) binding protein, an IL-1ß receptor, a dominant negative Toll/IL-1 signaling adapter, and CC-chemokine binding protein (MVAΔ4-HIV); (ii) harbor a deletion of an additional (fifth) viral gene, encoding uracil-DNA glycosylase (MVAΔ5-HIV); or (iii) represent the parental MVA backbone as a control (MVA-HIV). We performed head-to-head comparisons of the cellular and humoral immune responses that were elicited by these vectors during homologous prime-boost immunization regimens utilizing either high-dose (2 × 10(8) PFU) or low-dose (1 × 10(7) PFU) intramuscular immunization of rhesus macaques. At all time points, a majority of the HIV-specific T cell responses, elicited by all vectors, were directed against Env, rather than Gag, determinants, as previously observed with other vector systems. Both modified vectors elicited up to 6-fold-higher frequencies of HIV-specific CD8 and CD4 T cell responses and up to 25-fold-higher titers of Env (gp120)-specific binding (nonneutralizing) antibody responses that were relatively transient in nature. While the correlates of protection against HIV infection remain incompletely defined, our results indicate that the rational deletion of specific genes from MVA vectors can positively alter their cellular and humoral immunogenicity profiles in nonhuman primates.

Correlative study between neuron-specific enolase and blood sugar level in ischemic stroke patients.

BACKGROUND: A study to investigate the level of the neurobiochemical marker, Neuron-Specific Enolase (NSE), at the time of admission and its correlation with the blood sugar level in ischemic stroke patients. PATIENTS AND METHODS: We investigated 90 patients with complete stroke who were admitted to the Stroke Unit of the Department of Neurology at Sri Aurobindo Institute of Medical Sciences. NSE was measured with commercially available quantitative 'sandwich' enzyme-linked immunosorbent assay kits obtained from R and D Systems. Hyperglycemia was defined as blood glucose concentration ≥ 7 mmol / L, and measured using the glucose oxidase method immediately. RESULTS: Significantly increased NSE and lipid profile levels were found in ischemic stroke patients as compared to the control. Hyperglycemic ischemic stroke patients had increased levels of NSE, lipid profile, and National Institute of Health stroke scale scores (NIHSS score) compared to normoglycemic ischemic stroke patients. In addition the serum NSE level of hyperglycemic stroke patients was also positively correlated with the blood sugar level (r = 0.734 P < 0.001). CONCLUSIONS: Hyperglycemia predicts an increased risk of poor outcome after ischemic stroke and it is reflected by a significantly increased level of Neuron-Specific Enolase.

Plasmid profile and drug resistance pattern of zoonotic Salmonella isolates from Indian buffaloes.

BACKGROUND: Buffalo is the major source of animal protein in south-east Asia, including India; therefore, the presence of multiple drug resistance in Salmonella strains of buffalo meat and milk products is of immense public health concern. METHODOLOGY: Forty-six strains of Salmonella enterica subspecies enterica belonging to eight serovars (S. Anatum, 13; S. Weltevreden, 13; S. Rostock, 6; S. Typhimurium, 5; S. Gallinarum, 5; S. Stockholm, 1; S. Dublin, 1; and S. Orion, 2), isolated from buffalo meat and diseased buffaloes were studied for their antibiotic sensitivity and plasmid profile. RESULTS: All except six strains of Salmonella had one or more plasmids. Virulence plasmid of ~35MdA was present in 39 isolates while 19 strains had one to six additional plasmids with molecular weight ranging from 1 Mda > 35 Mda. A plasmid-free S. Anatum strain was resistant to seven drugs including fluoroquinolones, while strains having six to seven plasmids were resistant to fewer antimicrobial drugs. One S. Anatum isolate, resistant to 11 antibiotics, had only one plasmid. Eight serovars of Salmonella could be divided into 28 resistotypes on the basis of antimicrobial sensitivity assay. Most strains were resistant to streptomycin (84.8%) followed by kanamycin (58.7%), gentamicin (52.2%), ampicillin (50%) and oxytetracycline (50%). Few strains were resistant to cefotaxime (2.2%), amoxycillin (2.2%) and newer fluoroquinolones (6.5%). CONCLUSION: Multiple drug resistance was common among Salmonella isolates of buffalo origin, particularly against aminoglycosides, oxytetracycin, ampicillin and cephalexin. Presence of plasmids is not mandatory for occurrence of multiple drug resistance in S. enterica strains.

Phase I study of ipilimumab, an anti-CTLA-4 monoclonal antibody, in patients with relapsed and refractory B-cell non-Hodgkin lymphoma.

PURPOSE: The growth of non-Hodgkin lymphomas can be influenced by tumor-immune system interactions. Cytotoxic T-lymphocyte antigen 4 (CTLA-4) is a negative regulator of T-cell activation that serves to dampen antitumor immune responses. Blocking anti-CTLA-4 monoclonal antibodies improves host resistance to immunogenic tumors, and the anti-CTLA-4 antibody ipilimumab (MDX-010) has clinical activity against melanoma, prostate, and ovarian cancers. EXPERIMENTAL DESIGN: We did a phase I trial of ipilimumab in patients with relapsed/refractory B-cell lymphoma to evaluate safety, immunologic activity, and potential clinical efficacy. Treatment consisted of ipilimumab at 3 mg/kg and then monthly at 1 mg/kg x 3 months (dose level 1), with subsequent escalation to 3 mg/kg monthly x 4 months (dose level 2). RESULTS: Eighteen patients were treated, 12 at the lower dose level and 6 at the higher dose level. Ipilimumab was generally well tolerated, with common adverse events attributed to it, including diarrhea, headache, abdominal pain, anorexia, fatigue, neutropenia, and thrombocytopenia. Two patients had clinical responses; one patient with diffuse large B-cell lymphoma had an ongoing complete response (>31 months), and one with follicular lymphoma had a partial response lasting 19 months. In 5 of 16 cases tested (31%), T-cell proliferation to recall antigens was significantly increased (>2-fold) after ipilimumab therapy. CONCLUSIONS: Blockade of CTLA-4 signaling with the use of ipilimumab is well tolerated at the doses used and has antitumor activity in patients with B-cell lymphoma. Further evaluation of ipilimumab alone or in combination with other agents in B-cell lymphoma patients is therefore warranted.

Effect of increasing duration of diabetes mellitus type 2 on glycated hemoglobin and insulin sensitivity.

Non-insulin dependent diabetes mellitus (NIDDM) is the most rapidly growing chronic metabolic disorder in the world. With advancement in the age and duration of diabetes there is a gradual tendency for the level of blood sugar to rise along with a subsequent increase in the HbA1c as well as in the fasting insulin level. Whether this is an aging process or increased frequency of diabetes is still controversial. The correlation between glucose and insulin sensitivity is consistent with the idea that the degree of chronic hyperglycemia is a cause of excessive insulin resistance in type 2 diabetes, i.e. the insulin resistance which characterizes type 2 diabetes but not nondiabetic subjects matched for age, gender, family history and duration of diabetes. The study comprised a total of 76 subjects out of which 30 were normal, non-diabetic persons and the rest 46 were diabetics with different duration of time in years, after being diagnosed diabetic. Data was analyzed after dividing the subjects into four groups-Group 1 comprised of one year old diabetics, Group 2 was made up of those, who had diabetes, for the past 2-5 years, Group 3 included patients who were diabetic since more than 5 years and Group 4 included non-diabetics as the normal control group. The results obtained indicated that the HbA1c levels showed a significant increase with the duration of diabetes as well as the insulin level showed a significant correlation after adjustment for age, sex and duration of diabetes.

Study of adenosine deaminase activity in pulmonary tuberculosis and other common respiratory diseases.

Tuberculosis has, in a short span of time, become a major health problem in the third world or developing countries like India. In view of this, a retrospective study was conducted to study Adenosine deaminase activity in serum and pleural fluid in patients affected with Pulmonary Tuberculosis and other common non-tubercular chronic respiratory diseases. The study was carried out on 100 patients suffering from various pulmonary disorders, between January 2002 and August 2002. Thirty-five normal healthy individuals were included as control subjects. ADA estimation was done by sensitive colorimetric method. The study revealed that the serum ADA activity was higher in patients of tuberculous pulmonary and pleural diseases and non-tuberculous pulmonary diseases than in control subjects. The mean serum ADA activity in the patients' group was 35.5±6.93 u/l as compared to 16.20±2.85 u/l in control group, showing a highly significant (P≪0.001) difference. ADA activity was highest in tuberculous pleuropulmonary diseases. The pleural fluid ADA activity was higher in pyogenic pleural effusion than in tuberculous pleural effusion.

Serum total ganglioside level correlates with clinical course in melanoma patients after immunotherapy with therapeutic cancer vaccine.

The authors tested the hypothesis that the therapeutic destruction of residual tumors may be reflected in the level of serum total gangliosides (sTG). The sTG level was evaluated in 17 patients with in-transit melanoma and 70 patients with resected regional node metastasis, who have received a polyvalent therapeutic melanoma cell vaccine. The treatment response was determined by regression of in-transit metastases or by overall survival after resection. sTG levels were measured, blinded, before and after immunotherapy. The mean sTG level of the in-transit melanoma patients increased from 18.57 +/- 3.18 mg/dL pretreatment to 23.7 +/- 5.5 mg/dL between weeks 2 and 16 after initiation of treatment (p(2) < 0.0001). By week 24, the level had returned to its prevaccine level in the seven clinical responders (18.1 +/- 2.3 mg/dL vs. 20.4 +/- 3.2 mg/dL; p(2) < 0.050) but remained higher than its prevaccine level in the 10 nonresponders (23.3 +/- 5.1 mg/dl vs. 17.2 +/- 2.7 mg/dL). Similarly, the sTG level of the patients with nodal metastases increased between weeks 2 and 16 after the first vaccine treatment; by week 24, it had returned to pretreatment level in patients who survived more than 56 weeks but remained significantly elevated (p(2) < 0.01) in patients who survived less than 56 weeks. The sTG level increased between weeks 2 and 16 in all vaccine recipients and returned to prevaccine level by week 24 in all who showed measurable regression of in-transit melanoma (7 of 17 patients) or improved overall survival (53 of 70 patients). The data suggest that sTG level could be a potential tool for assessing the response to immunotherapy in melanoma patients by week 24.