Evaluation of 2 Novel Ratio-Based Metrics for Lumbar Spinal Stenosis.

BACKGROUND AND PURPOSE: Quantitative metrics of the dural sac such as the cross-sectional area are commonly used to evaluate central canal stenosis. The aim of this study was to analyze 2 new metrics to measure spinal stenosis on the basis of the ratio between the dural sac and disc cross-sectional areas (DDRCA) and the dural sac and disc anterior-posterior diameters (DDRDIA) and compare them with established quantitative metrics of the dural sac. MATERIALS AND METHODS: T2-weighted axial MR images (n = 260 patients) were retrospectively evaluated, graded for central canal stenosis as normal (no stenosis), mild, moderate, or severe from L1/L2 through L5/S1 with 1 grade per spinal level and annotated to measure the DDRCA and DDRDIA. Thresholds were obtained using a decision tree classifier on a subset of patients (n = 130) and evaluated on the remaining patients (n = 130) for accuracy and consistency across demographics, anatomic variation, and clinical outcomes. RESULTS: DDRCA and DDRDIA had areas under the receiver operating characteristic curve of 98.6 (97.4-99.3) and 98.0 (96.7-98.9) compared with dural sac cross-sectional area at 96.5 (95.0-97.7) for binary classification. DDRDIA and DDRCA had κ scores of 0.75 (0.71-0.79) and 0.80 (0.75-0.83) compared with dural sac cross-sectional area at 0.62 (0.57-0.66) for multigrade classification. No significant differences (P > .1) in the area under the receiver operating characteristic curve were observed for the DDRDIA across variations in the body mass index. The DDRDIA also had the highest area under the receiver operating characteristic curve among symptomatic patients (visual analog scale ≥ 7) or patients who underwent surgery. CONCLUSIONS: Ratio-based metrics (DDRDIA and DDRCA) are accurate and robust to anatomic and demographic variability compared with quantitative metrics of the dural sac and better correlated with symptomatology and surgical outcomes.

STAT3 inhibition prevents lung inflammation, remodeling, and accumulation of Th2 and Th17 cells in a murine asthma model.

BACKGROUND: STAT3 drives development of Th17 cells and cytokine production by Th2 and Th17 cells, which contribute to asthma. Alternative asthma treatments are needed, especially for the Th17 phenotype. We sought to determine whether C188-9, a small-molecule STAT3 inhibitor, can block Th2 and Th17 cell expansion and cytokine production to prevent house dust mite (HDM)-induced airway inflammation and remodeling. METHODS: Three groups of C57BL/6 mice were treated intranasally (IN) and intraperitoneally (IP) daily for 3 weeks with the following: (i) vehicle 1 IN and vehicle 2 IP, (ii) HDM IN and vehicle 2 IP, or (iii) HDM IN and C188-9 IP. Sections of lung were stained with Alcian Blue/PAS and examined microscopically. Total (t) STAT3, STAT3 phosphorylated on Y705 (pSTAT3), IL-17, IL-13, IL-5, and IL-4 levels were measured in lung protein extracts and serum using Luminex beads. Frequencies of Th2-type and Th17-type lymphocytes were assessed in lungs and bronchoalveolar lavage fluid (BALF) by multiparametric flow cytometry. RESULTS: HDM inhalation markedly increased airway goblet cell numbers and thickness of the epithelium and subepithelial smooth muscle layer, which was accompanied in the whole lung by increased pSTAT3, IL-4, IL-5, IL-13, and IL-17, and % CD4+ T cells that produce IL-5, IL-13, and IL-17. HDM inhalation also increased serum IL-4 and IL-17 levels and increased BALF % CD4+ T cells that produce IL-5 and IL-13. Remarkably, treatment with C188-9 normalized each endpoint. CONCLUSION: HDM-induced airway inflammation, remodeling, and Th2/Th17-type cell accumulation involve STAT3 activation that can be prevented by C188-9 treatment.

A mix of S and ΔS variants of STAT3 enable survival of activated B-cell-like diffuse large B-cell lymphoma cells in culture.

Activated B-cell-like diffuse large B-cell lymphoma (ABC DLBCL) is characterized by increased expression and activator of signal transducer and activator of transcription 3 (STAT3). ABC DLBCL cells require STAT3 for growth in culture. In ABC DLBCL cells, eosinophils and perhaps all cells, four variant STAT3 mRNAs (Sα, ΔSα, Sß and ΔSß) are present as a result of two alternative splicing events, one that results in the inclusion of a 55-residue C-terminal transactivation domain (α) or a truncated C-terminal domain with 7 unique residues (ß) and a second that includes (S) or excludes (ΔS) the codon for Ser-701 in the linker between the SH2 and C-terminal domains. A substantial literature indicates that both α and ß variants are required for optimal STAT3 function, but nothing is known about functions of ΔS variants. We used a knockdown/re-expression strategy to explore whether survival of ABC DLBCL cells requires that the four variants be in an appropriate ratio. No single variant rescued survival as well as STAT3Sα-C, Sα with activating mutations (A661C and N663C) in the SH2 domain. Better rescue was achieved when all four variants were re-expressed or Sα and ΔSα or Sß and ΔSß were re-expressed in pairs. Rescue correlated with expression of STAT3-sensitive genes NFKBIA and NFKBIZ. We consider a variety of explanations why a mix of S and ΔS variants of STAT3 should enable survival of ABC DLBCL cells.

Drug-repositioning screening identified piperlongumine as a direct STAT3 inhibitor with potent activity against breast cancer.

Signal transducer and activator of transcription (STAT) 3 regulates many cardinal features of cancer including cancer cell growth, apoptosis resistance, DNA damage response, metastasis, immune escape, tumor angiogenesis, the Warburg effect and oncogene addiction and has been validated as a drug target for cancer therapy. Several strategies have been used to identify agents that target Stat3 in breast cancer but none has yet entered into clinical use. We used a high-throughput fluorescence microscopy search strategy to identify compounds in a drug-repositioning library (Prestwick library) that block ligand-induced nuclear translocation of Stat3 and identified piperlongumine (PL), a natural product isolated from the fruit of the pepper Piper longum. PL inhibited Stat3 nuclear translocation, inhibited ligand-induced and constitutive Stat3 phosphorylation, and modulated expression of multiple Stat3-regulated genes. Surface plasmon resonance assay revealed that PL directly inhibited binding of Stat3 to its phosphotyrosyl peptide ligand. Phosphoprotein antibody array analysis revealed that PL does not modulate kinases known to activate Stat3 such as Janus kinases, Src kinase family members or receptor tyrosine kinases. PL inhibited anchorage-independent and anchorage-dependent growth of multiple breast cancer cell lines having increased pStat3 or total Stat3, and induced apoptosis. PL also inhibited mammosphere formation by tumor cells from patient-derived xenografts. PL's antitumorigenic function was causally linked to its Stat3-inhibitory effect. PL was non-toxic in mice up to a dose of 30 mg/kg/day for 14 days and caused regression of breast cancer cell line xenografts in nude mice. Thus, PL represents a promising new agent for rapid entry into the clinic for use in treating breast cancer, as well as other cancers in which Stat3 has a role.

Effect of copper- and zinc-methionine supplementation on bioavailability, mineral status and tissue concentrations of copper and zinc in ewes.

The effect of feeding Cu- and Zn-methionine to ewes was studied in a 240d feeding trial. The plasma and tissue Cu and Zn concentrations and Cu/Zn-superoxide dismutase (Cu/Zn-SOD) activity were employed to assess the relative bioavailability from Cu- and Zn-methionine. The macro and micronutrient intake, utilization, plasma mineral status, tissue accumulation of Cu and Zn as well as wool concentration of Cu and Zn were studied in ewes (n=12) fed a corn-soybean meal based basal diet with 50% more Cu and Zn supplementation over the basal diet either from Cu- and Zn-sulfate (Cu-Sulf+Zn-Sulf group) or Cu- and Zn-methionine (Cu-Meth+Zn-Meth group). The average daily feed intake and body weight gain of ewes did not differ due to dietary supplementation of Cu- and Zn-methionine. However, dry matter intake was comparatively lower and thus resulted in better feed: gain in Cu- and Zn-methionine group as compared to ewes fed Cu- and Zn-sulfate. Supplementation of Cu and Zn over the basal diet either from methionine-chelated or sulfate sources resulted in increased plasma Cu and Zn as well as Cu/Zn-SOD activity on d-30, which indicated a positive correlation between plasma Cu and Zn and Cu/Zn-SOD activity. The gut absorption, liver concentrations of Cu and Zn, and liver Cu/Zn-SOD activity were significantly (P<0.01) higher in ewes supplemented with Cu- and Zn-methionine compared to Cu- and Zn-sulfate. Periodical analysis of wool samples indicated no significant difference in Cu and Zn content between Cu-and Zn-methionine and Cu- and Zn-sulfate groups. Feeding of Cu and Zn from methionine-chelated source resulted in reduced (P<0.01) excretion of Cu and Zn in feces indicating their better utilization, and this will have positive implication on environment. The gut absorption values, plasma and liver tissue concentrations of Cu and Zn supported the hypothesis that Cu- and Zn-methionine supplements have better bioavailability compared to Cu- and Zn-sulfate and Cu- and Zn-dependent enzyme (Cu/Zn-SOD) could be used to determine the bioavailability of Cu and Zn.

Human genetic variation studies and HLA class II loci.

Human leucocyte antigen (HLA) loci widely known for their role in generation of immune responses by encoding cell-surface heterodimers are often considered to be effective for the purpose of reconstructing human phylogenies due to high degree of polymorphism and less recombination. In the present study, we have made an attempt to study HLA class II loci (DRB1, DQA1 and DQB1) in inferring phylogenetic relationship based on both phylogenetic and haplotype approach. In the phylogenetic approach, the compiled database of 19 populations got segregated and finely resolved in three basal clusters with very high bootstrap values corresponding to four geo-ethnic groups of Africans, Orientals, Americans and Caucasians. Maximum- likelihood phylogram has placed North Indian Hindus alongside other Caucasian populations. Haplotype analysis revealed high range of haplotype diversity with nearly 144 observed haplotypes. The haplotype distribution suggested that numbers of Caucasian-specific haplotypes are frequently found among north Indian Hindus. Our results indicate that if the property of less recombination is explored to assign extended haplotypes, followed by strong interpretation based on more logistic statistical model, then there is a high possibility that HLA class II loci can infer exact and accurate phylogenetic assessments as revealed by mtDNA and Y-chromosome markers.

DNA short tandem repeat profiling of three North Indian populations.

POPULATION: Fifty healthy unrelated individuals were randomly chosen from each of the three populations viz., Bhargavas, Chaturvedies, and Brahmins. Three generation pedigree charts were prepared to ensure sirname endogamy in Bhargavas Chaturvedies and group endogamy in Brahmins subjects were chosen from several parts of Uttar Pradesh, a northern state of the Indian republic.

Central cholinergic mechanism for control of urine volume and electrolyte excretion.

1. Studies were conducted on urine volume and electrolyte excretion chiefly of Na and K in anaesthetized hydrated dogs. 2. Central injection of acetylcholine caused a dose dependent antidiuretic response but without any change in excretion of urinary Sodium (UNa) and Potassium (UK). 3. After central atropinization, antidiuretic response to acetylcholine was partially blocked without any effect on electrolyte excretion. 4. Intracerebroventricularly (I.C.V.) administered acetylcholine after vagotomy and spinalectomy, each done separately and together also elicited an antidiuretic response. There was no effect on electrolyte excretion. 5. It is thus suggested that acetylcholine may be acting on central cholinergic receptors concerned with A.D.H. release.

Effect of centrally administered glucagon on urine output in dogs.

The effect of minimal doses of glucagon, administered by intracerebroventricular (ICV) and intracisternal(IC) routes, on urine output in mongrel dogs have been studied. The dose of 2.0 mug of glucagon was found to be the minimal dose for diuresis on peripheral administration. This dose when centrally administered, produced an oliguric effect in animals. This effect was not observed in vagosympathectomised-spinal cord transectomised or adrenalectomised animals. It is suggested that the probable efferents might be the sympathetic fibres as they are present in vagi nerves as well in the spinal cord (26). The observations made in an attempt to find out the organ responsible for the oliguric effect, showed that some substance released from the adrenal cortex has an influence on the kidney's excretory function.

Central component of the cario-accelerator action of angiotensin II in dogs.

Injection of angiotensin II into a lateral cerebral ventricle (I.C.V.) or into a peripheral vein of anaesthetized dog elicited a rise in blood pressure and transient bradycardia followed by sustained tachycardia. Spinal transection at C2 and bilateral vagotomy abolished the central cardiovascular effect of I.C.V. angiotensin. However, in spinal transected dogs the usual pressor response to intravenous angiotensin was observed. Since the transient bradycardia was absent in bilaterally vagotomized dogs or in dogs with their blood pressure stabilized by means of a mechanical buffer devise it must be reflex in origin. The tachycardia was more marked in vagotomized dogs. Prior administration of a beta adrenergic receptor blocking agent propranolol, blocked the tachycardia, but the pressor response was unaffected. The cardiovascular responses to centrally administered angiotensin were practically abolished by prior treatment of dogs with reserpine or by extirpation of both adrenal glands. Thus it may be concluded that ICV angiotensin induces a centrogenic release of catecholamines from the adrenal medulla which is responsible for the cardiovascular responses.