Inset techniques for the DIEP flap - what improves aesthetic outcomes?

INTRODUCTION: The deep inferior epigastric perforator (DIEP) flap is widely considered as the gold standard in breast reconstruction. The inset technique of the DIEP flap is crucial in determining the overall aesthetic outcome; however, to date no systematic review is available that comprehensively assesses the various techniques. Evaluation of topic: A systematic review was performed according to the PRISMA guidelines. The methodology is outlined within our published protocol (Prospero CRD42023449477). Included articles met a minimal criterion compromising of the intervention (DIEP free flap for breast reconstruction) and outcomes (aesthetic and clinical outcomes). Six articles were included in this review, with a total of 346 patients and a follow-up ranging from 6 months to 4 years. Four articles were of a prospective case series study design, one article was a randomized controlled trial, and one article was a case-control study. The risk of bias was assessed to be high in the case series, but low and moderate in the randomized controlled trial and case-control study respectively. CONCLUSION: Although limited by the quality of the evidence, the single aesthetic unit principle, dual-plane inset, elimination of the need for a skin paddle, appropriate flap positioning and rotation, and algorithmic in-setting may all improve the aesthetic outcome of DIEP free flaps.

Evidence-based identification and characterization of methicillin-resistant Staphylococcusaureus isolated from subclinical mastitis in dairy buffaloes of Pakistan.

Background: Methicillin-resistant Staphylococcus aureus (MRSA), affecting livestock and human beings, has become a global public health hazard with economic consequences. Aims: The current study was designed to investigate the prevailing MRSA-associated subclinical mastitis and associated risk factors in dairy buffaloes. The study also highlighted the genetic variations and in silico-based proteomic differences among MRSA isolates. Methods: Out of 516 milk samples, 45.93% (237/516) were found positive for subclinical mastitis, while the prevalence of S. aureus was recorded 56.12%. The methicillin resistance in S. aureus isolates was evaluated by oxacillin disc diffusion test and molecular identification of the mecA gene. Results: The results revealed a phenotypic and molecular prevalence of MRSA at 45.11% and 18.79%, respectively. The risk factor analysis revealed that among various assumed risk factors, parity, milking hygiene, milker care during milking, milk yield, housing system, and floor type were significantly associated with subclinical mastitis in buffaloes. The sequencing and phylogenetic analysis showed no significant genetic variations among study isolates and depicted a high similarity with isolates from Africa, USA, India, Italy, Turkey, and Iran. The in-silico protein analysis showed that all sequences had the same protein motifs resembling penicillin protein 2a except Buff-13, whose protein structure resembles alpha-catenin-like protein hmp-1. Conclusion: The current study was the first report of the genotypic characterization and in silico protein analysis of MRSA from dairy buffaloes in Pakistan. The result highlighted the importance of antimicrobial resistance (AMR) and development of control strategies against MRSA infections.

Molecular evidence, risk factors analysis, and hematological alterations associated with Theileria spp. spillover in captive wild mouflon sheep in Punjab, Pakistan.

Background: Landscape anthropization and interaction between domestic and wild animals are the major contributing factors involved in the emergence of new pathogens in wild animals. Theileriosis is an emerging issue of wild ungulates, especially in the tropical and subtropical areas of the globe. Aims: The current study investigated the mouflon sheep for Theileria infection using molecular methods and hematological analysis. Methods: This study was conducted on a total of 103 captive wild mouflon sheep present in eight different recreational zoos, and wildlife parks in Punjab, Pakistan to investigate the genotypic prevalence of Theileria spp. by targeting 18S rRNA and molecular evidence for Theileria spillover between domestic and wild mouflon sheep by phylogenetic analysis. The association of assumed risk factors and the effect of Theileria spp. on various hematological parameters were also assessed. Results: The results depicted that Theileria spp. was prevalent in 8 (7.77%, CI 95%: 3.99-14.59%), and 11 (10.68%, CI 95%: 06.07-18.12%) animals based on microscopy, and PCR, respectively. The phylogenetic analysis of the 18S rRNA gene of Theileria spp. from mouflon revealed a close resemblance with T. annulata from domestic animals. The risk factor analysis revealed that tick infestation, enclosure hygiene, previous tick infestation history, and the presence of wooden logs in the enclosure were significantly (P<0.05) associated with the occurrence of Theileria spp. infection in the captive mouflon sheep of Pakistan. Furthermore, a significant reduction in blood parameters like PCV, RBCs count, Hb, and platelets was observed in Theileria-positive animals. Conclusion: This study is the first evidence at the molecular level to characterize the spillover of Theileria spp. between the captive wild mouflon sheep and domestic animals of Pakistan, and it will be useful in developing control strategies for emerging theileriosis in captive wild animals.

Ambulatory screening of diabetic neuropathy and predictors of its severity in outpatient settings.

INTRODUCTION: Diabetic neuropathy is one of the most common causes of chronic neuropathic symptomatology and the most disabling and difficult-to-treat diabetic microangiopathic complication. The neuropathies associated with diabetes are typically classified into generalized, focal and multifocal varieties. There exists a scarcity of literature studying the correlation of different patient- and disease-related variables with severity of neuropathy. OBJECTIVES: This study aims to delineate the prevalence of diabetic neuropathy in type 2 diabetes, describe its characteristics and find out predictors of its severity. MATERIAL AND METHODS: Eight hundred consecutive diabetic patients presenting to outpatient department (OPD) of Khan Research Labs (KRL) General Hospital and Centre for Diabetes and Liver diseases, Islamabad, during March-June, 2015 were made to complete a self-administered questionnaire (Michigan Neuropathy Screening Instrument-MNSI) and underwent a thorough physical examination according to MNSI protocols. A score of >2 was considered to be diagnostic for DPN. Patient and disease variables were noted. MNSI score was used as an index of severity of diabetic peripheral neuropathy (DPN). Correlation of several patient- and disease-related variables with the severity of DPN was determined using multivariate regression. RESULTS: Out of a total 800 patients screened, 90 (11.25%) were found to have diabetic neuropathy. Of these 90, 45.5% were males, the median age was 54.47 ± 10.87 years and the median duration of diabetes was 11.12 ± 9.8 years. The most common symptom was found to be numbness (63.6%) followed by generalized body weakness (61.5%). The common findings on physical examination were dry skin/callus (38.7%) and deformities (14.7%). Duration of diabetes was found to be the strongest predictor for development and severity of diabetic neuropathy followed by glycemic controls (HbA1c values) and age. CONCLUSION: Duration of diabetes rather than diabetic controls predicts better the development and severity of diabetic neuropathy indicating a failure of intensive treatment to avert such complications.

An enzymatic method for the detection of human serum albumin.

Albumin is the most abundant protein in human serum. A dye-binding method is commonly used in clinical laboratories for its estimation using different types of dyes. However, all these dye methods were interfered by a variety of compounds. Here we present a method for the detection of albumin in human serum and other biological fluids. The principle is based on the fact that lactate dehydrogenase isoenzyme-5 (LDH-5) binds specifically to Dextran-Blue (DB). Albumin inhibits the binding of LDH-5 with DB. Absence of LDH activity in DB fraction after gel filtration indicates the presence of albumin in sample and vice versa.

An endogenous inhibitor of Ca++-ATPase from human placenta.

Intracellular free calcium is regulated by Ca(++)-ATPase, one form present on the plasma membrane (PM Ca(++)-ATPase) and the other on sarcoplasmic (endoplasmic) reticulum (SR/ER Ca(++)-ATPase). An endogenous inhibitor of SR Ca(++)-ATPase from human placenta was shown to be present in normal placenta and the activity was not detectable in placenta from preeclamptic patients. The inhibitor was distributed in cytosol and microsomes. The inhibition of Ca(++)-ATPase by this inhibitor was concentration- and time-dependent. The inhibitor neither bound to DEAE- nor CM-sepharose resins at pH 7.5 and 8.5. Furthermore, it was heat stable for 15 min up to 55 degrees C and completely destroyed at 80 degrees C in a few minutes. It was also observed to be stable at room temperature for at least 3 months. The purification and characterization of this inhibitor would be valuable in achieving an understanding of the normal regulation of Ca(++)-ATPase in the placenta during pregnancy.

GroEL protects the sarcoplasmic reticulum Ca(++)-dependent ATPase from inactivation in vitro.

The molecular chaperone, GroEL, facilitates correct protein folding and inhibits protein aggregation. The function of GroEL is often, though not invariably, dependent on the co-chaperone, GroES, and ATP. In this study it is shown that GroEL alone substantially reduces the inactivation of purified Ca(++)-ATPase from rabbit skeletal muscle sarcoplasmic reticulum. In the absence of GroEL, the enzyme became completely inactive in about 45-60 hours when kept at 25 degrees C, while in the presence of an equimolar amount of GroEL, the enzyme remained approximately 80% active even after 75 hours. Equimolar amounts of BSA or lysozyme were unable to protect the enzyme from inactivation under identical conditions. Analysis by SDS-PAGE showed GroEL was acting by blocking the aggregation of ATPase at 25 degrees C. GroEL was not as effective in protection at -20 degrees C or 4 degrees C. These results are discussed in the context of current models of the GroEL mechanism.

Inhibition studies on LDH isoenzyme purified from Uromastix testes.

An LDH isoenzyme was purified to homogeneity from uromastix testes and its inhibition spectrum towards known LDH isoenzyme inhibitors studied. Platinum compounds inhibited the enzyme in the forward reaction (pyruvate-->lactate) only, n-hexanediol and colchicine showed no inhibition and gossypol acetic acid (GAA) strongly inhibited both the forward and reverse reactions and the reactions were time-dependent. Oxalate caused non-competitive inhibition (Ki app = IC50 = 0.15 mM) of the forward reaction, NADH was more effective in blocking inhibition by GAA than pyruvate. This enzyme was also unable to use ketocaproic acid as a substrate.

Purification and properties of lactate dehydrogenase from liver of Uromastix hardwickii.

Lactate dehydrogenase isoenzyme-1 was purified from liver of Uromastix hardwickii using colchicine-Sepharose and heat-inactivation methods. The crude enzyme showed four isoenzymes by agarose gel electrophoresis (AGE). The purified enzyme showed a single band after native AGE and SDS-PAGE corresponding to a molecular weight of 34 kDa. The enzyme did not bind with DEAE-Sepharose at pH 7.2. The optimum pH for forward reaction was 7.5, while for reverse reaction, the maximum activity was at pH 9.5. The Km values for pyruvate, NADH, lactate and NAD+ were 0.105, 0.045, 9.0 and 0.011 mM, respectively. The pyruvate showed maximum activity at about 150 microM and then starts showing inhibition at higher concentration. Pre-heating of enzyme showed that it was stable at 80 degrees C for 30 min and at 100 degrees C it became inactive immediately. Oxalate, glutamate, Cu2+, Co2+, Mn2+, and Mg2+ have shown inhibitory effects both for forward- and reverse-reactions. From these properties, we suggest that LDH-1 from Uromastix liver may be quite different from that of other vertebrates.