A new midline closure technique without skin sutures: Long-term outcomes of primary repair of pilonidal sinus disease.

BACKGROUND: Currently, the focus regarding pilonidal sinus disease is put on the treatment techniques. The aim of the study is to compare postoperative long-term complications and recurrence of two surgical techniques. MATERIAL AND METHODS: From February 2015 to December 2020, male patients with pilonidal sinus disease attended at two general surgery outpatient centers were randomly assigned to either Group 1 (n=80; excision and primary closure) or Group 2 (n=80; excision and midline closure without skin sutures). Patients with recurrent or complicated pilonidal sinus or with prior surgical procedures were excluded from the study. Intergroup postoperative results and recurrence throughout the follow-up period were analyzed. RESULTS: Significant decrease (p<0.001) in the duration of the surgical procedure (35 to 25 minutes), length of hospital stay (one day to the day of the surgery), and of the time required to return to work (15 to 12 days) was seen for Group 2 patients. The complication rate (wound infection and seroma) was lower in Group 2 compared to Group 1 (n = 3; 3.7% vs n = 10; 12.5%; p = 0.014). During the five-year mean follow-up, five patients (6.2%) in Group 1 had recurrence compared to none in Group 2 (p = 0.023). CONCLUSIONS: Midline primary closure method without skin sutures - easy to learn and implement and has no complication or recurrence in the long-term follow-up - may be an ideal method in cases where excision and primary repair is planned, especially in patients with sinus orifices located in the midline.

SEVA 4.0: an update of the Standard European Vector Architecture database for advanced analysis and programming of bacterial phenotypes.

The SEVA platform (https://seva-plasmids.com) was launched one decade ago, both as a database (DB) and as a physical repository of plasmid vectors for genetic analysis and engineering of Gram-negative bacteria with a structure and nomenclature that follows a strict, fixed architecture of functional DNA segments. While the current update keeps the basic features of earlier versions, the platform has been upgraded not only with many more ready-to-use plasmids but also with features that expand the range of target species, harmonize DNA assembly methods and enable new applications. In particular, SEVA 4.0 includes (i) a sub-collection of plasmids for easing the composition of multiple DNA segments with MoClo/Golden Gate technology, (ii) vectors for Gram-positive bacteria and yeast and [iii] off-the-shelf constructs with built-in functionalities. A growing collection of plasmids that capture part of the standard-but not its entirety-has been compiled also into the DB and repository as a separate corpus (SEVAsib) because of its value as a resource for constructing and deploying phenotypes of interest. Maintenance and curation of the DB were accompanied by dedicated diffusion and communication channels that make the SEVA platform a popular resource for genetic analyses, genome editing and bioengineering of a large number of microorganisms.

Automated design and implementation of a NOR gate in Pseudomonas putida.

Boolean NOR gates have been widely implemented in Escherichia coli as transcriptional regulatory devices for building complex genetic circuits. Yet, their portability to other bacterial hosts/chassis is generally hampered by frequent changes in the parameters of the INPUT/OUTPUT response functions brought about by new genetic and biochemical contexts. Here, we have used the circuit design tool CELLO for assembling a NOR gate in the soil bacterium and the metabolic engineering platform Pseudomonas putida with components tailored for E. coli. To this end, we capitalized on the functional parameters of 20 genetic inverters for each host and the resulting compatibility between NOT pairs. Moreover, we added to the gate library three inducible promoters that are specific to P. putida, thus expanding cross-platform assembly options. While the number of potential connectable inverters decreased drastically when moving the library from E. coli to P. putida, the CELLO software was still able to find an effective NOR gate in the new chassis. The automated generation of the corresponding DNA sequence and in vivo experimental verification accredited that some genetic modules initially optimized for E. coli can indeed be reused to deliver NOR logic in P. putida as well. Furthermore, the results highlight the value of creating host-specific collections of well-characterized regulatory inverters for the quick assembly of genetic circuits to meet complex specifications.

Contextual dependencies expand the re-usability of genetic inverters.

The implementation of Boolean logic circuits in cells have become a very active field within synthetic biology. Although these are mostly focussed on the genetic components alone, the context in which the circuit performs is crucial for its outcome. We characterise 20 genetic NOT logic gates in up to 7 bacterial-based contexts each, to generate 135 different functions. The contexts we focus on are combinations of four plasmid backbones and three hosts, two Escherichia coli and one Pseudomonas putida strains. Each gate shows seven different dynamic behaviours, depending on the context. That is, gates can be fine-tuned by changing only contextual parameters, thus improving the compatibility between gates. Finally, we analyse portability by measuring, scoring, and comparing gate performance across contexts. Rather than being a limitation, we argue that the effect of the genetic background on synthetic constructs expands functionality, and advocate for considering context as a fundamental design parameter.

A Standardized Inverter Package Borne by Broad Host Range Plasmids for Genetic Circuit Design in Gram-Negative Bacteria.

Genetically encoded logic gates, especially inverters-NOT gates-are the building blocks for designing circuits, engineering biosensors, or decision-making devices in synthetic biology. However, the repertoire of inverters readily available for different species is rather limited. In this work, a large whole of NOT gates that was shown to function previously in a specific strain of Escherichia coli, was recreated as broad host range (BHR) collection of constructs assembled in low, medium, and high copy number plasmid backbones of the SEVA (Standard European Vector Architecture) collection. The input/output function of each of the gates was characterized and parametrized in the environmental bacterium and metabolic engineering chassis Pseudomonas putida. Comparisons of the resulting fluorescence cytometry data with those published for the same gates in Escherichia coli provided useful hints on the portability of the corresponding gates. The hereby described inverter package (20 different versions of 12 distinct gates) borne by BHR plasmids thus becomes a toolbox of choice for designing genetic circuitries in a variety of Gram-negative species other than E. coli.

Are synthetic biology standards applicable in everyday research practice?

The issue of standardization in synthetic biology is a recurring one. As a discipline that incorporates engineering principles into biological designs, synthetic biology needs effective ways to communicate results and allow different researchers (both academic and industrial) to build upon previous results and improve on existing designs. An aspect that is left out of the discussions, especially when they happen at the level of academic and industrial consortia or policymaking, is whether or not standards are applicable or even useful in everyday research practice. In this caucus article, we examine this particular issue with the hope of including it in the standardization discussions agenda and provide insights into a topic that synthetic biology researchers experience daily.

Pathways to cellular supremacy in biocomputing.

Synthetic biology uses living cells as the substrate for performing human-defined computations. Many current implementations of cellular computing are based on the "genetic circuit" metaphor, an approximation of the operation of silicon-based computers. Although this conceptual mapping has been relatively successful, we argue that it fundamentally limits the types of computation that may be engineered inside the cell, and fails to exploit the rich and diverse functionality available in natural living systems. We propose the notion of "cellular supremacy" to focus attention on domains in which biocomputing might offer superior performance over traditional computers. We consider potential pathways toward cellular supremacy, and suggest application areas in which it may be found.

Effects of weapon types, interventions, and transport times on complications in combat injuries to musculoskeletal system.

OBJECTIVE: To analyze the effects of weapon type, medical interventions, and transportation time on complications due to combat-related injuries of the musculoskeletal and soft tissue (ST). METHODS: A total of 108 patients treated at the emergency department (ED) over a period of 3.5 years were included in this study. The effects of weapon type, type of first intervention team, interventions at the ED, and transport time on complications were compared by retrospectively assessing patients' recorded data. RESULTS: The average age of 108 patients with ST injuries was 24 years. 25 patients developed complications. The ratio of complications in patients with injuries from explosive weapons was 25.42% and was significantly greater (p<0.05) than the rate observed from non-explosive weapons. Regarding the transport time of patients reaching the hospital, the greatest complication rate was 30.77% in patients transported to the hospital in 2-4 h; this rate was significantly (p<0.05) greater than for those transported in other time ranges. Regarding the ED procedures, the complication rate was 37.50% and significantly higher (p<0.05) in the group that received debridement + irrigation + late primary repair. CONCLUSION: Transporting the injured to the ED within the first 2 h, treatment including irrigation and debridement, and secondary healing following explosive injuries containing contamination and dead tissue appear to be good options for reducing complications.

Experiences of Conflict Zone-Related Ballistic Renal Injury.

To analyze our approaches and experiences in patients with renal injuries accompanying penetrating abdominal injuries admitted to the hospital due to high kinetic energy weapons. Patients including renal injuries associated with penetrating abdominal injuries due to gunshot wounds and fragments of shell treated at our institution between February 2002 and May 2013 were retrospectively analyzed. Total 21 patients were included in this study, 20 of whom (86.2 %) were male and 1 (13.8 %) female. Renal injury was scaled in 2 patients as grade 1, 4 patients as grade 2, 4 patients as grade 3, 8 patients as grade 4, and 3 patients as grade 5. While conservative treatment was applied to patients with grade 1 and 2 injuries, 2 patients with grade 3 injury underwent renal repair and the other 13 patients underwent nephrectomy. The amount of blood transfused in all cases was determined to be mean 8.28 (6-16) units for red blood cells (ES) and 9 (6-17) units for fresh frozen plasma. When patients were assessed according to the Penetrating Abdominal Trauma Index (PATI) scores, the median score and average PATI score were 35 and 37.6 (10-70), respectively. A physical examination along with a quick assessment of vital signs should be made, and unnecessary and time-consuming investigations such as abdominal tomography and angiographic intervention should be avoided. Conservative and organ preservation should be considered absolutely for appropriately selected patients, namely in stable patients in whom kinetic energy transfer is less and who have bullet path away from midline. However, the majority of these patients are considered to be hemodynamically unstable condition, possible complications of the procedure to be applied, and selection of patients should be carefully evaluated.

An Integrated System for Precise Genome Modification in Escherichia coli.

We describe an optimized system for the easy, effective, and precise modification of the Escherichia coli genome. Genome changes are introduced first through the integration of a 1.3 kbp Landing Pad consisting of a gene conferring resistance to tetracycline (tetA) or the ability to metabolize the sugar galactose (galK). The Landing Pad is then excised as a result of double-strand breaks by the homing endonuclease I-SceI, and replaced with DNA fragments bearing the desired change via λ-Red mediated homologous recombination. Repair of the double strand breaks and counterselection against the Landing Pad (using NiCl2 for tetA or 2-deoxy-galactose for galK) allows the isolation of modified bacteria without the use of additional antibiotic selection. We demonstrate the power of this method to make a variety of genome modifications: the exact integration, without any extraneous sequence, of the lac operon (~6.5 kbp) to any desired location in the genome and without the integration of antibiotic markers; the scarless deletion of ribosomal rrn operons (~6 kbp) through either intrachromosomal or oligonucleotide recombination; and the in situ fusion of native genes to fluorescent reporter genes without additional perturbation.

Reliability of the mangled extremity severity score in combat-related upper and lower extremity injuries.

BACKGROUND: Decision of limb salvage or amputation is generally aided with several trauma scoring systems such as the mangled extremity severity score (MESS). However, the reliability of the injury scores in the settling of open fractures due to explosives and missiles is challenging. Mortality and morbidity of the extremity trauma due to firearms are generally associated with time delay in revascularization, injury mechanism, anatomy of the injured site, associated injuries, age and the environmental circumstance. The purpose of the retrospective study was to evaluate the extent of extremity injuries due to ballistic missiles and to detect the reliability of mangled extremity severity score (MESS) in both upper and lower extremities. MATERIALS AND METHODS: Between 2004 and 2014, 139 Gustillo Anderson Type III open fractures of both the upper and lower extremities were enrolled in the study. Data for patient age, fire arm type, transporting time from the field to the hospital (and the method), injury severity scores, MESS scores, fracture types, amputation levels, bone fixation methods and postoperative infections and complications retrieved from the two level-2 trauma center's data base. Sensitivity, specificity, positive and negative predictive values of the MESS were calculated to detect the ability in deciding amputation in the mangled limb. RESULTS: Amputation was performed in 39 extremities and limb salvage attempted in 100 extremities. The mean followup time was 14.6 months (range 6-32 months). In the amputated group, the mean MESS scores for upper and lower extremity were 8.8 (range 6-11) and 9.24 (range 6-11), respectively. In the limb salvage group, the mean MESS scores for upper and lower extremities were 5.29 (range 4-7) and 5.19 (range 3-8), respectively. Sensitivity of MESS in upper and lower extremities were calculated as 80% and 79.4% and positive predictive values detected as 55.55% and 83.3%, respectively. Specificity of MESS score for upper and lower extremities was 84% and 86.6%; negative predictive values were calculated as 95.45% and 90.2%, respectively. CONCLUSION: MESS is not predictive in combat related extremity injuries especially if between a score of 6-8. Limb ischemia and presence or absence of shock can be used in initial decision-making for amputation.

[The relationship between Injury Severity Scores and transfusion requirements of 108 consecutive cases injured with high kinetic energy weapons: a tertiary center end-mode mortality analysis]. / Yüksek kinetik enerjili silahlarla yaralanmis ardisik 108 olguya ait ISS degerleri ile transfüzyon gereksinimleri arasindaki iliski ve ileri merkez son mod mortalite analizi.

BACKGROUND: We aimed in this study to investigate the relationship between Injury Severity Score (ISS) and transfusion strategies required during medical intervention in patients wounded by high kinetic energy (HKE) gunshot, and to analyze end-mode mortality. METHODS: The medical data of patients were included in the study. We evaluated whether there was any significant correlation in terms of demographic characteristics, HKE weapon type, ISSs, and transfusion strategy options and transfusion requirements. RESULTS: Causes of mortality in cases resulting in mortality during hospitalization were evaluated. One hundred and eight consecutive patients were included in the study. All patients except one were male, with an average age of 25 years. 64.8% of them were injured by long-barreled firearms, whereas 35.2% were injured by explosives. Average ISS was 13.9. ISS values for the patients with and without transfusion were 16 (5-48) and 9 (3-36), respectively. Causes of mortality were evaluated in terms of systemic inflammatory response syndrome (SIRS), sepsis, and multiorgan dysfunction syndrome (MODS). DISCUSSION: It was determined that there was a significant correlation between increase in ISS values in cases with HKE weapon wounds and their transfusion requirements, whereas this requirement was independent of the ISS value in cases with explosive wounds.

[Analysis of the pediatric trauma score in patients wounded with shrapnel; the effect of explosives with high kinetic energy: results of the first intervention center]. / Pediatrik travma skorunun yüksek kinetik enerjiye sahip parça etkili yaralanmalardaki analizi: Ilk müdahale merkezi sonuçlari

BACKGROUND: We aimed to assess the pediatric trauma score analysis in pediatric trauma cases due to shrapnel effect of explosives material with high kinetic energy. METHODS: The data of 17 pediatric injuries were reviewed retrospectively between February 2002 and August 2005. The information about age, gender, trauma-hospital interval, trauma mechanism, the injured organs, pediatric Glasgow coma score (PGCS), pediatric trauma score (PTS), hemodynamic parameters, blood transfusion, interventions and length of hospital stay (LHS) were investigated. RESULTS: While all patients suffered from trauma to the extremities, only four patients had traumatic lower-limb amputation. Transportation time was <=1 hour in 35% of cases, and >1 hour in 65% of cases. While PTS was found as <=8 in 35.3% of cases (n=6), the score was found to be higher than 8 in 64.7% of them (n=11). Median heart rate in patients with PTS <=8 was 94 beats/min. This value was 70 beats/min in those with PTS >8 (p=0.007). Morbidity rates of PTS <=8 cases and PTS >8 cases were 29.4% and 5.9%, respectively (p=0.026). While LHS was 22.8 days in PTS <=8 cases, LHS was found to be only 4 days in PTS >8 cases. This difference was found to be statistically significant (p=0.001). CONCLUSION: PTS is very efficient and a time-saving procedure to assess the severity of trauma caused by the shrapnel effect. The median heart rate, morbidity, and LHS increased significantly in patients with PTS <=8.

[The affecting factors on the complication ratio in abdominal gunshot wounds]. / Karin içi atesli silah yaralanmalarinda komplikasyon oranlarini etkileyen faktörler.

BACKGROUND: We aimed to investigate the affecting factors on the complication ratio in abdominal gunshot wounds. METHODS: Twenty-one patients with abdominal gunshot wounds were analyzed between February 2002 and May 2005. The effects of the interval between trauma and presentation to the hospital, the number of injured abdominal and extra-abdominal organs, penetrating abdominal trauma index (PATI), and blood transfusion were evaluated. RESULTS: 90.4% of all patients were transported to the hospital and underwent their first evaluation in the first two hours. The complication rate was 7.1% in patients who had <3 injured organs and 71% in the others (p<0.0001). 71.4% of the patients had isolated abdominal trauma, while 28.6% had additional extra-abdominal organ trauma. The complication rate was 7.7% in 13 patients with PATI score <25 and 62.5% in 8 patients with a PATI score ≥25 (p<0.0001). In 10 patients who underwent blood transfusion of ≥3 units, the complication rate was 50% (p<0.0001). CONCLUSION: In our study, PATI score, multiple blood transfusions and the number of injured intra-abdominal organs were the most important factors affecting the rate of postoperative complications in penetrating abdominal gunshot wounds. We found that the interval between trauma and presentation to the hospital and number of injured extra-abdominal organs did not affect the complication rate.

Controlled viable release of selectively captured label-free cells in microchannels.

Selective capture of cells from bodily fluids in microchannels has broadly transformed medicine enabling circulating tumor cell isolation, rapid CD4(+) cell counting for HIV monitoring, and diagnosis of infectious diseases. Although cell capture methods have been demonstrated in microfluidic systems, the release of captured cells remains a significant challenge. Viable retrieval of captured label-free cells in microchannels will enable a new era in biological sciences by allowing cultivation and post-processing. The significant challenge in release comes from the fact that the cells adhere strongly to the microchannel surface, especially when immuno-based immobilization methods are used. Even though fluid shear and enzymes have been used to detach captured cells in microchannels, these methods are known to harm cells and affect cellular characteristics. This paper describes a new technology to release the selectively captured label-free cells in microchannels without the use of fluid shear or enzymes. We have successfully released the captured CD4(+) cells (3.6% of the mononuclear blood cells) from blood in microfluidic channels with high specificity (89% ± 8%), viability (94% ± 4%), and release efficiency (59% ± 4%). We have further validated our system by specifically capturing and controllably releasing the CD34(+) stem cells from whole blood, which were quantified to be 19 cells per million blood cells in the blood samples used in this study. Our results also indicated that both CD4(+) and CD34(+) cells released from the microchannels were healthy and amenable for in vitro culture. Manual flow based microfluidic method utilizes inexpensive, easy to fabricate microchannels allowing selective label-free cell capture and release in less than 10 minutes, which can also be used at the point-of-care. The presented technology can be used to isolate and purify a broad spectrum of cells from mixed populations offering widespread applications in applied biological sciences, such as tissue engineering, regenerative medicine, rare cell and stem cell isolation, proteomic/genomic research, and clonal/population analyses.

Comparison of modified darn repair and Lichtenstein repair of primary inguinal hernias.

OBJECTIVE: This study was designed to compare the results of the Modified Darn Repair through Lichtenstein procedure in inguinal hernias. MATERIALS AND METHODS: The study involved 322 patients with inguinal hernia, operated in General Surgery Departments of Gulhane Military Medical Academy and Sirnak Military Hospital between 1998 and 2004. The durations of operation time, hospitalization, and time to return to daily activities and postoperative complication and recurrence rates were evaluated. Lichtenstein procedure was applied on 170 patients (Group 1), and modified darn repair was applied on 152 patients (Group 2). RESULTS: The average follow-up period was 56 months. For the Lichtenstein procedure, the average duration of operation was 56 min; the average time to return to routine activities was 20 days. The number of patients with postoperative complications was 20 (11.7%), and the number of patients with recurrence was 1 (0.6%). For modified darn repair, the average duration of operation was 48 min; the time to return to daily activities was 20 days. The number of patients with postoperative complication was three (1.9%), and no recurrences were noted. The hospitalization time of the groups was similar. DISCUSSION: Modified darn repair is a reliable method for inguinal hernia repair with short hospitalization time, low rate of postoperative complications, and recurrence.

Increased intra-abdominal pressure causes bacterial translocation in rabbits.

BACKGROUND: Abdominal compartment syndrome (ACS) is defined as intra-abdominal hypertension associated with organ dysfunction. Subsequently, increased intra-abdominal pressure (IAP) adversely affects the pulmonary, cardiovascular, renal, musculoskeletal/integumentary and central nervous systems. Bacterial translocation (BT), which is defined as the movement of viable enteric bacteria to the mesenteric lymph nodes, liver and spleen, occurs after various types of stress and results in splanchnic ischemia. In this experimental study, we aimed to investigate the effects of various levels of increased IAP on BT in rabbits; IAP was increased by the intra-abdominal balloon-insufflation method, thus simulating noncompliant abdominal-wall closure under tension. METHODS: Fifty rabbits were randomly assigned to 1 of 5 groups, with each group comprising 10 animals. In group I (control group), an intra-abdominal balloon was placed without inflation. In groups II, III, IV and V, IAPs of 10, 15, 20 and 25 mmHg, respectively, were established via inflation of the intra-abdominal balloon. All groups underwent laparotomy after 12 hours. Multiple biopsies were taken from ileocecal lymph nodes, the spleen and liver. RESULTS: BT was observed to some degree in all 4 experimental groups. A gradual increase in the phenomenon was noted as IAP increased from 10 to 15 mmHg; and BT was overt at 20 mmHg, and significant at 25 mmHg. Klebsiella pneumoniae, Serratia marcescens, and Escherichia coil were the predominant pathogens identified by culture. CONCLUSION: We propose that an intravesical pressure (IVP) of 15 mmHg is the critical point for BT in patients with increased IAP. In this experimental study, BT occurred when IVP reached 20 mmHg. We suggest that IVP monitoring is desirable in the management of patients with ACS, and that decompressive laparotomy should be performed in patients with IVP >20 mmHg.

Enteric and rectal duplications and duplication cysts in the adult.

Alimentary tract duplication and duplication cysts are rare congenital malformations. The ileum is the most frequently affected site. However, alimentary tract duplication and duplication cysts can occur at any point along the gastrointestinal tract. Early diagnosis and prompt surgical treatment is the best way to prevent associated morbidity. This article presents the cases of three patients admitted to Gulhane Military Medical Academy with signs of acute abdomen, intra-abdominal mass and chronic abdominal pain. These patients were found to have enteric duplication, duplication cyst and/or retro-rectal cyst. The literature on alimentary tract duplications is reviewed.