Tissue expansion: Concepts,techniques and unfavourable results

The phenomenon of tissue expansion is observed in nature all the time. The same properties of the human skin to stretch and expand and yield extra skin if placed under continuous stress over a prolonged period of time has been utilised for reconstructive purposes with the help of a silicon balloon inserted under the skin and progressively filled with saline. The technique of tissue expansion is now more than three decades old and has been a value addition to our armamentarium in reconstructive surgery in all parts of the body. However, it still requires careful patient selection, meticulous planning and faultless execution to successfully carry out the process, which usually lasts for more than 8-12 weeks and involves two sittings of surgery. Any compromise in this process can lead to unfavourable results and complications, some minor, which allow continuance of the process to attain the expected goal and others major, which force abandonment of the process without reaching the expected goal. This article seeks to highlight the intricacies of the concept of tissue expansion, the technique related to flawless execution of the process and likely complications with emphasis on their management. We also present our results from a personal series of 138 patients operated over a period of 18 years between 1994 and 2012.KEY WORDS: Complications, tissue expansion, unfavourable results     

Ovarian Regulation of Kisspeptin Neurones in the Arcuate Nucleus of the Rhesus Monkey (Macaca mulatta)

Tonic gonadotrophin secretion throughout the menstrual cycle is regulated by the negative‐feedback actions of ovarian oestradiol (E₂) and progesterone. Although kisspeptin neurones in the arcuate nucleus (ARC) of the hypothalamus appear to play a major role in mediating these feedback actions of the steroids in nonprimate species, this issue has been less well studied in the monkey. In the present study, we used immunohistochemistry and in situ hybridisation to examine kisspeptin and KISS1 expression, respectively, in the mediobasal hypothalamus (MBH) of adult ovariectomised (OVX) rhesus monkeys. We also examined kisspeptin expression in the MBH of ovarian intact females, and the effect of E₂, progesterone and E₂ + progesterone replacement on KISS1 expression in OVX animals. Kisspeptin or KISS1 expressing neurones and pronounced kisspeptin fibres were readily identified throughout the ARC of ovariectomised monkeys but, on the other hand, in intact animals, kisspeptin cell bodies were small in size and number and only fine fibres were observed. Replacement of OVX monkeys with physiological levels of E₂, either alone or with luteal phase levels of progesterone, abolished KISS1 expression in the ARC. Interestingly, progesterone replacement alone for 14 days also resulted in a significant down‐regulation of KISS1 expression. These findings support the view that, in primates, as in rodents and sheep, kisspeptin signalling in ARC neurones appears to play an important role in mediating the negative‐feedback action of E₂ on gonadotrophin secretion, and also indicate the need to study further their regulation by progesterone.     

A novel dihydro phenylquinazolinone-based two-in-one colourimetric chemosensor for nickel(ii), copper(ii) and its copper complex for the fluorescent colourimetric nanomolar detection of the cyanide anion

Currently, considerable efforts have been devoted to the detection and quantification of hazardous multi-analytes using a single probe. Herein, we have developed a simple, environment-friendly colourimetric sensor for the sensitive, selective and rapid detection of Ni²⁺ and Cu²⁺ ions using a simple organic Schiff base ligand L in methanol–Tris–HCl buffer (1 : 1 v/v, 10 mM, pH = 7.2). The probe L exhibited a binding-induced colour change from colourless to yellow and fluorescence quenching in the presence of both Ni²⁺ and Cu²⁺ ions. The interactions between L and the respective metal ions were studied by Job''s plot, electrospray ionisation-mass spectrometry (ESI-MS), Fourier-transform infrared spectroscopy (FT-IR), density functional theory (DFT) and time-dependent density functional theory (TDDFT) calculations. The limit of detection (LOD) of L towards Ni²⁺ and Cu²⁺ was calculated to be 7.4 × 10⁻⁷ M and 4.9 × 10⁻⁷ M, respectively. Furthermore, the L–Cu²⁺ complex could be used as a new cascade fluorescent-colourimetric sensor to detect CN⁻ ions with a very low level of detection (40 nM). Additionally, L could operate in a wide pH range, and thus was successfully applied for the detection and quantification of Ni²⁺ and Cu²⁺ in environmental samples, and for building OR- and IMPLICATION-type logic gates.

A novel colorimetric chemosensor L has been developed for the detection of Ni²⁺ and Cu²⁺ ions. The obtained L–Cu²⁺ complex can be used as a new cascade fluorescent-colorimetric sensor for the nanomolar detection of CN⁻ ions. This chemosensor has practical application.     

Diagnostic Imaging Use for the Initial Evaluation of Low Back Pain by Primary Care Providers in the United States: 2011-2016

ObjectiveHigh-value care guidelines from multiple medical societies recommend against imaging for the initial evaluation of low back pain in the absence of red flag symptoms. We aimed to determine the current temporal and geographic landscape of imaging ordering patterns for this indication among US primary care providers.MethodsUsing a national commercial insurance claims database, we identified patients between 18 and 64 years old who presented to a primary care provider for an initial evaluation of low back pain between 2011 and 2016. Patients were identified via International Classification of Diseases codes, and the use of diagnostic imaging was identified by Current Procedural Terminology codes. Geographic regions were based on the location of patient residence.ResultsOverall, 627,118 encounters met inclusion criteria. Imaging acquisitions increased over time, from 14% of encounters in 2011 to 16% in 2016 (P < .01). Radiographs represented 96% of ordered imaging, CT 2%, and MRI 3%. The likelihood of having any imaging for low back pain varied significantly by US census region and by US state (P < .01). The greatest use of imaging was in the Midwest (13.9%) and the South (18.5%), and lowest in the Northeast and West (6.2% and 13.6%).DiscussionImaging utilization for the initial evaluation of low back pain by primary care providers has increased on a national level from 2011 to 2016, largely represented by radiographs. Significant regional variation also exists. Encouragingly, the use of advanced imaging has remained at a low level in the primary care setting (<1.0%).     

Diminished degradation of myelin basic protein by anti-sulfatide antibody and interferon-gamma in myelin from glia maturation factor-deficient mice

In this study we show the effect of anti-sulfatide (RmAb) antibodies and inflammatory cytokines, TNF-alpha and IFN-gamma in inducing myelin basic protein (MBP) degradation in myelin isolated from control wild type (WT) and glia maturation factor (GMF)-deficient (GMF-KO) mice. GMF was not detected in isolated myelin from WT and GMF-KO mice although it is present in brains of WT mice. Our results show that calcium-dependent neutral protease activity caused significantly elevated degradation of 18.5 and/or 17.5kDa isoforms of MBP in WT myelin treated with RmAb or IFN-gamma. In contrast, MBP degradation in isolated myelin from GMF-KO mice remained unaffected following treatment with RmAb, IFN-gamma, or GM-CSF. Neither the 14kDa isoform of MBP nor proteolipid protein (PLP) showed an elevated degradation compared to controls. A virtual absence of GM-CSF, TNF-alpha and IFN-gamma in GMF-KO brain compared to WT was also evident when the animals were challenged with MOG 35-55. Additionally, the myelin from GMF-KO mice showed difference in distribution of myelin oligodendrocyte glycoprotein (MOG) and beta-tubulin in a sucrose density gradient myelin-axolemmal fractions compared to WT. Taken together, our data suggests a role for GMF in the biochemical organization of myelin and thereby its effect on MBP degradation induced by RmAb and IFN-gamma.     

Conjugation to Ascorbic Acid Enhances Brain Availability of Losartan Carboxylic Acid and Protects Against Parkinsonism in Rats

Identification of renin-angiotensin system in the interplay of hypertension and neurodegeneration has paved the way for the repurposing of antihypertensive drugs against Parkinsonism. Losartan carboxylic acid (LCA), the potent AT1 blocker metabolite of losartan, suffers from poor bioavailability and brain access. Since ascorbate transporters have earlier shown enough flexibility as carriers, we have conjugated losartan carboxylic acid to ascorbic acid with the aim of achieving higher oral/brain availability. Ester of LCA and ascorbic acid (FED) was developed keeping in view the substrate specificity of ascorbate transporters. Oral/brain bioavailability was assessed using in vivo pharmacokinetic model. Effect on central nervous system (CNS) and protection against Parkinsonism was evaluated using in vivo models. FED enhanced bioavailability of LCA. The higher brain availability of LCA enabled CNS protection as evident from the increase in locomotor activity, improved motor coordination, and protection against drug-induced catatonia. In conclusion, FED offers an approach to repurpose LCA against Parkinsonism. This can encourage further investigation to simultaneously address hypertension and neurodegeneration.     

Quantitation and characterization of glutathionyl haemoglobin as an oxidative stress marker in chronic renal failure by mass spectrometry

OBJECTIVES: Glutathionyl haemoglobin (GS-Hb) belonging to the class of glutathionylated proteins has been investigated as a possible marker of oxidative stress in different chronic diseases. The purpose of this study was to examine whether glutathionyl haemoglobin can serve as an oxidative stress marker in non-diabetic chronic renal failure patients on different renal replacement therapies (RRT) through its quantitation, and characterization of the specific binding site of glutathione in haemoglobin molecule by mass spectrometric analysis. DESIGN AND METHODS: The study group consisted of non-diabetic chronic renal failure patients on renal replacement therapy (RRT): hemodialysis (HD), continuous ambulatory peritoneal dialysis (CAPD) and renal allograft transplant (Txp) patients. Haemoglobin samples of these subjects were analyzed by liquid chromatography electrospray ionization mass spectrometry for GS-Hb quantitation. Characterization of GS-Hb was done by tandem mass spectrometry. Levels of erythrocyte glutathione (GSH) and lipid peroxidation (as thiobarbituric acid reacting substances) were measured spectrophotometrically, while glycated haemoglobin (HbA1c) was measured by HPLC. RESULTS: GS-Hb levels were markedly elevated in the dialysis group and marginally in the transplant group as compared to the controls. GS-Hb levels correlated positively with lipid peroxidation and negatively with the erythrocyte glutathione levels in RRT groups indicating enhanced oxidative stress. De novo sequencing of the chymotryptic fragment of GS-Hb established that glutathione is attached to Cys-93 of the beta globin chain. Mass spectrometric quantitation of total glycated haemoglobin showed good agreement with HbA1c estimation by conventional HPLC method. CONCLUSIONS: Glutathionyl haemoglobin can serve as a clinical marker of oxidative stress in chronic debilitating therapies like RRT. Mass spectrometry provides a reliable analytical tool for quantitation and residue level characterization of different post-translational modifications of haemoglobin.