Annotated Whole-Genome Multilocus Sequence Typing Schema for Scalable High-Resolution Typing of Streptococcus pyogenes.

Streptococcus pyogenes is a major human pathogen with high genetic diversity, largely created by recombination and horizontal gene transfer, making it difficult to use single nucleotide polymorphism (SNP)-based genome-wide analyses for surveillance. Using a gene-by-gene approach on 208 complete genomes of S. pyogenes, a novel whole-genome multilocus sequence typing (wgMLST) schema was developed, comprising 3,044 target loci. The schema was used for core-genome MLST (cgMLST) analyses of previously published data sets and 265 newly sequenced draft genomes with other molecular and phenotypic typing data. Clustering based on cgMLST data supported the genetic heterogeneity of many emm types and correlated poorly with pulsed-field gel electrophoresis macrorestriction profiling, superantigen gene profiling, and MLST sequence type, highlighting the limitations of older typing methods. While 763 loci were present in all isolates of a data set representative of S. pyogenes genetic diversity, the proposed schema allows scalable cgMLST analysis, which can include more loci for an increased resolution when typing closely related isolates. The cgMLST and PopPUNK clusters were broadly consistent in this diverse population. The cgMLST analyses presented results comparable to those of SNP-based methods in the identification of two recently emerged sublineages of emm1 and emm89 and the clarification of the genetic relatedness among isolates recovered in outbreak contexts. The schema was thoroughly annotated and made publicly available on the chewie-NS online platform (https://chewbbaca.online/species/1/schemas/1), providing a framework for high-resolution typing and analyzing the genetic variability of loci of particular biological interest.

Genomics and proteomics approaches to the study of cancer-stroma interactions.

BACKGROUND: The development and progression of cancer depend on its genetic characteristics as well as on the interactions with its microenvironment. Understanding these interactions may contribute to diagnostic and prognostic evaluations and to the development of new cancer therapies. Aiming to investigate potential mechanisms by which the tumor microenvironment might contribute to a cancer phenotype, we evaluated soluble paracrine factors produced by stromal and neoplastic cells which may influence proliferation and gene and protein expression. METHODS: The study was carried out on the epithelial cancer cell line (Hep-2) and fibroblasts isolated from a primary oral cancer. We combined a conditioned-medium technique with subtraction hybridization approach, quantitative PCR and proteomics, in order to evaluate gene and protein expression influenced by soluble paracrine factors produced by stromal and neoplastic cells. RESULTS: We observed that conditioned medium from fibroblast cultures (FCM) inhibited proliferation and induced apoptosis in Hep-2 cells. In neoplastic cells, 41 genes and 5 proteins exhibited changes in expression levels in response to FCM and, in fibroblasts, 17 genes and 2 proteins showed down-regulation in response to conditioned medium from Hep-2 cells (HCM). Nine genes were selected and the expression results of 6 down-regulated genes (ARID4A, CALR, GNB2L1, RNF10, SQSTM1, USP9X) were validated by real time PCR. CONCLUSIONS: A significant and common denominator in the results was the potential induction of signaling changes associated with immune or inflammatory response in the absence of a specific protein.

Treatment of caustic ingestion: an analysis of 239 cases.

The objective of the present study was to analyze a 37-year historical series of patients who had ingested caustic substances, and who were treated in a teaching hospital, to assess the effectiveness of the therapy administered during this period. We studied 239 patients who ingested caustic soda (NaOH) from 1957 to 1994. Data were collected from the medical records of the patients and from interviews with them and analyzed by software and by statistical tests of association. The results showed that more women than men ingested caustic substances (57%, n=153). Ingestion was associated with suicidal intent in 60% of cases and was accidental in 37.2% of cases. The amount of substance ingested ranged from a trace to as much as three tablespoons, with the amount tending to be larger in the suicide attempts. Of the 215 patients for whom information about complications due to ingestion was available, 88.4% (190) presented lesions of the esophagus (73% with stenosis), 1% died during the acute phase, and 10.6% did not present complications. The data revealed that the presence and severity of stenosis were correlated with the amount of caustic substance ingested. The treatment received by the patients in the study sample varied over the years according to the prevailing literature recommendations. Based on our review, we conclude that neither the use of an antidote nor early treatment immediately after ingestion is effective. Treatment with a corticosteroid (1.5-2 mg/kg/day prednisone), an antibiotic, and a high-protein and hypercaloric diet seems to be beneficial for patients who ingest small or medium amounts of caustic soda. When 2-3 tablespoons are ingested, corticosteroids, in addition to being unable to prevent the formation of esophageal stenosis, increase the risk of other complications.

Blindness after laryngectomy and bilateral neck dissection in a diabetic patient: case report.

CONTEXT: Neck dissection that accompanies resection of the primary lesion in malignant tumors of the upper aerodigestive tracts may cause complications inherent to the procedure or to prolongation of surgical time, increasing the risks for the patient. Among the complications that might occur is blindness, a rare complication with only 10 cases reported in the literature thus far. OBJECTIVE: To present the case of a diabetic patient submitted to total laryngectomy and modified and selective neck dissection that resulted in blindness. CASE REPORT: The authors report on a patient submitted to total laryngectomy and selective neck dissection on the left side, and modified radical neck dissection on the right, who developed blindness. This was probably due to intraoperative hypotension plus the contribution of decompensated diabetes mellitus and thrombosis of the internal jugular vein on the right side. The possible causes, risk factors and care to be taken to prevent this rare but highly debilitating complication are discussed.

Time necessary for neovascularization of a tracheal segment by the sternohyoid muscle.

INTRODUCTION: The reimplantation of a complete tracheal segment (CTS) as an autograft or homograft often results in total or partial necrosis of the CTS. However, previous experiments suggested that when the CTS is first vascularized by the sternohyoid muscle (SM) 21 days before the reimplantation, the CTS can be reimplanted without ischemia or stenosis. However, it is not clear if the CTS requires 21 days to attain full revascularization. GOAL: This study aims to ascertain the minimum time necessary for revascularization of the 6 rings CTS by the SM allowing reimplantation free of ischemia. METHODS: Fifteen mongrel dogs, divided into 3 groups were submitted to 2 sequential surgical procedures. In the first procedure, a SM flap was used to envelop 6 rings CTS in all dogs. After periods of 21 days (group 1, n = 5), 14 days (group 2, n = 5) and 7 days (group 3, n = 5), the compound flaps (CTS and SM) were neovascularized. Each CTS was transected and mobilized from the trachea and then replaced again in its original site, as an autoflap. RESULTS: After a period of at least 60 days, none of dogs presented any respiratory alteration. Macroscopic and microscopic analyses demonstrated adequate viability and no stenosis. CONCLUSION: Our findings suggest that it is possible to neovascularize the CTS using a SM flap within a period as short as 1 week.

Ingestion of caustic substances and its complications.

CONTEXT: Caustic substances cause tissue destruction through liquefaction or coagulation reactions and the intensity of destruction depends on the type, concentration, time of contact and amount of the substance ingested. OBJECTIVES: To analyze the complications in patients who ingested caustic substances and correlate them with the amount of caustic soda ingested. DESIGN: Retrospective study. SETTING: University hospital, a referral center. PARTICIPANTS: A total of 239 patients who ingested caustic soda. MAIN MEASUREMENTS: The amount of granulated caustic substance ingested was measured as tablespoonfuls and the following complications were analyzed: esophagitis, esophageal stenosis and progression to cancer, fistulas, perforations, stomach lesions, brain abscesses, and death. Stenosis was classified as mild, moderate or severe according to the radiological findings. RESULTS: We observed an 89.3 % incidence of esophagitis; 72.6 % of the cases involved progression to stenosis and 1 per cent died during the acute phase. Stenosis was mild in 17.6 % of cases, moderate in 59.3 % and severe in 23 %. The incidence of stenosis was 80.8 per cent in women and 62.5 % in men. The incidence of stenosis was 46.9 % in the group that ingested "fragments" and 93.6 % in the group that ingested one or more tablespoonfuls of caustic substances. Among subjects who ingested one or more tablespoonfuls, 32.2 % developed lesions of the stomach-duodenum, whereas the ingestion of "fragments" was not sufficient to induce these lesions. There was no correlation between the intensity of lesions of the esophagus and of the stomach. Progression to cancer of the esophagus occurred in 1.8 % of cases, death during the chronic phase in 1.4 %, perforations in 4.6 %, fistulas in 0.9 %, and brain abscesses in 1.4 %. CONCLUSIONS: The complications were related to the amount of caustic soda ingested. Small amounts caused esophagitis or stenosis and large amounts increased the risk of fistulas, perforations and death.

[Intraesophageal pressure during esophageal speech in laryngectomized patients rehabilitated or no rehabilitated for oral communication]. / Pressão intra-esofágica durante a produção da voz esofágica em pacientes laringectomizados com e sem recuperação da capacidade de comunicação oral.

BACKGROUND: After laryngectomy for treatment of pharyngeal/laryngeal carcinomas the patients may be rehabilitated, for oral communication, with the esophageal speech. AIM: To study the intra-esophageal pressure during the esophageal speech. PATIENTS AND METHODS: It was measured the intra-esophageal pressure in 25 laryngectomized patients aged 40 to 70 years (median 57 years), 10 rehabilitated with esophageal speech and 15 unable to do so. The manometric method with continuous perfusion was used. The esophageal pressures was measured 3 to 5 cm below the upper esophageal sphincter when the patients tried to speak the vowel "a". Sometimes the air swallowed went to the stomach, with a peristaltic or simultaneous contraction in the esophageal body. RESULTS: During the attempt of esophageal speech the intra-esophageal pressure was higher in patients able to have esophageal speech (26.4 +/- 10.1 mm Hg, mean +/- SD) than in patients unable to do so (13.7 +/- 7.2 mm Hg). The esophageal contraction after a swallow of air was also higher in patients with esophageal speech (45.3 +/- 8.6 mm Hg) than in patients unable to do so (33.8 +/- 13.1 mm Hg). CONCLUSION: Laryngectomized patients rehabilitated with esophageal speech has a higher intra-esophageal pressure during speech than patients unable to do so, what may be consequence of the capacity to retain air inside the esophagus.

Effect of gastroesophageal reflux on hypertrophy of the base of the tongue.

The objective of this study was to determine a possible association between pharyngolaryngeal signs of gastroesophageal reflux (GER) and hypertrophy of the lymphoid follicles at the base of the tongue (HBT). For this purpose, 306 patients submitted to videolaryngoscopy were studied retrospectively and classified according to the presence and size of follicles on the base of the tongue. HBT was considered to be present when the follicles prevented the view of the vallecula. The patients were grouped according to the presence of videolaryngoscopic signs of GER and further subdivided according to the presence of esophageal, pharyngolaryngeal, and esophagopharyngolaryngeal symptoms, with an attempt made to relate these symptoms to the presence of HBT. HBT was detected in 62.4% (63/101) of the patients with signs of GER and in 29.3% (60/205) of patients with no signs of GER. When HBT incidence was studied according to the symptoms reported by the patient, the condition was found to be present in 57.2% (8/14) of patients with exclusively esophageal symptoms, in 63.6% (21/33) with esophagopharyngolaryngeal symptoms, and in 75% (15/20) with exclusively pharyngolaryngeal symptoms. We conclude therefore that HBT is associated with GER.

Use of a platysma myocutaneous flap for the reimplantation of a severed ear: experience with five cases.

CONTEXT: The traumatic loss of an ear greatly affects the patient because of the severe aesthetic deformity it entails. The characteristic format of the ear, with a fine skin covering a thin and elastic cartilage, is not found anywhere else in the human body. Thus, to reconstruct an ear, the surgeon may try to imitate it by sculpting cartilage and covering it with skin. OBJECTIVE: To use a platysma myocutaneous flap for the reimplantation of a severed ear in humans. DESIGN: Case report. SETTING: Emergency unit of the university hospital, Faculty of Medicine, Ribeirão Preto - USP. CASE REPORT: Five cases are reported, with whole ear reimplantation in 3 of them and only segments in 2 cases. The surgical technique used was original and was based on the principle of auricular cartilage revascularization using the platysma muscle. We implanted traumatically severed auricular cartilage into the platysma muscle. The prefabricated ear was later transferred to its original site in the form of a myocutaneous-cartilaginous flap. Of the 5 cases treated using this technique, 4 were successful. In these 4 cases the reimplanted ears showed no short- or long-term problems, with an aesthetic result quite close to natural appearance. In one case there was necrosis of the entire flap, with total loss of the ear. The surgical technique described is simple and utilizes the severed ear of the patient. Its application is excellent for skin losses in the auricular region or for the ear itself, thus obviating the need for microsurgery or the use of protheses or grafts.

Evaluation of esophageal motility in laryngectomized patients.

Laryngectomy for treatment of laryngeal-pharyngeal carcinomas may impair the sensation in the larynx and epiglottis, with consequent impairment of esophageal motility. Our aim in the present study was to investigate the esophageal motility of laryngectomized patients. Esophageal manometry was performed on 17 patients submitted to laryngectomy 2 to 71 months (median 29 months) before the examination. Eleven were rehabilitated with esophageal voice and six could not speak. Ten swallows of a 5 ml bolus of water were recorded at the lower esophageal sphincter and at 5, 10 and 15 cm above it. The lower esophageal sphincter pressure was measured by the rapid pull-through method and the upper esophageal sphincter pressure by the station pull-through method. The results were compared with those obtained for a control group of 40 healthy volunteers. The amplitude of contractions was lower and the number of nonperistaltic contractions was higher in laryngectomized patients than in volunteers (P < 0.05). The duration of lower esophageal sphincter relaxation (7.4 +/- 1.5 s) was shorter in laryngectomized patients than in volunteers (8.8 +/- 1.6 s, P < 0.05). The upper esophageal sphincter pressure was lower (34.9 +/- 29.1 mm Hg) in laryngectomized patients than in volunteers (61.2 +/- 20.8 mm Hg, P < 0.05). There was no difference between groups in contraction duration or velocity, in the numbers of multipeaked or failed contractions, lower esophageal sphincter pressure or in the number of swallows followed by complete lower esophageal sphincter relaxation. In conclusion, laryngectomy causes esophageal motility impairment characterized by low contraction amplitude, nonperistaltic contraction and shorter lower esophageal sphincter relaxation duration.

Esophageal motility in patients with esophageal caustic injury.

OBJECTIVE: There are few studies of esophageal function subsequent to the ingestion of lye. We investigated the esophageal motility of patients who had ingested liquid sodium hydroxide. METHODS: Esophageal manometry was performed on 21 patients who [1-53 yr before the manometric examination (median: 13 yr)] had drunk 10-30 g of sodium hydroxide diluted in water. The results were compared with those obtained for a control group of 22 healthy volunteers. RESULTS: The lower esophageal sphincter (LES) pressure of the caustic group (14.9 +/- 1.7 mm Hg, mean +/- SE) did not differ (p > 0.05) from that of the control group (17.4 +/- 1.1 mm Hg). LES pressure in eight patients was below 10 mm Hg. The amplitude of contraction was lower (p < 0.01) in the caustic group than in the control group. In the proximal part of the esophageal body, the duration of contraction was longer (p < 0.01) in the caustic group (2.3 +/- 0.1 s) than in controls (1.8 +/- 0.1 s). Nonperistaltic contraction was a finding in 14 patients, repetitive in five of them. The velocity of peristaltic contractions was higher (p < 0.05) in the caustic group (distal: 3.2 +/- 0.2 cm/s) than in the control group (2.2 +/- 0.2 cm/s). Although there was some impairment of esophageal motor function in 71% of the patients, they were asymptomatic when we performed esophageal manometry. CONCLUSIONS: Esophageal motility impairment was present in most of the patients who ingested sodium hydroxide. Nonperistaltic contractions of low amplitude were found most frequently.

Tracheal neovascularization: a method involving mobilization of a complete tracheal neovascularized segment using a sternohyoid muscle flap.

Tracheal segmental free grafts always tend to undergo necrosis with consequent occlusion of the airway. Revascularized grafts are impossible to carry out, since the trachea is devoid of a major vascular pedicle that would permit microvascular reconstruction. On the other hand, neovascularized grafts carry a potential for success but have not been sufficiently studied. Neovascularization of a six-ring circumferential tracheal segment (CTS) was studied in dogs using a sternohyoid muscle (SM) flap. Three different procedures were carried out. In group 1 the six-ring CTS was inside a free graft. In group 2 the SM was freed from its proximal connection and rotated to wrap a corresponding six-ring CTS; it was then sutured and left in place for 21 days. After this period it was again approached, and the six-ring CTS was sectioned and sutured back in place, leaving the distally pedicled SM untouched. In group 3 an identical procedure was carried out, but the SM flap was left with a proximally rather than a distally based flap. All surviving animals were followed up for at least 1 year, and the results were analyzed by clinical and tracheoscopic observations and by macroscopic and microscopic studies after the animals were killed. All animals in group 1 died within 18 days; the studies showed necrosis and occlusion of the CTS. In groups 2 and 3 there was no degenerative change of the CTS, whose aspect was close to normal on macroscopic and microscopic examination. We conclude that the free CTS graft is totally inviable. In contrast, neovascularization of the CTS occurs when the segment is first wrapped around with an SM flap. This ensures CTS viability and opens new perspectives for homotransplantation.

Rupture of the aorta due to a malpositioned tracheal cannula in a 4-month-old baby.

Complications of tracheostomy are not rare and are even more common in children. They may be acute or chronic and sometimes may have serious consequences. We report here a rare situation in which a malpositioned tracheal cannula in a false tract from the neck to the mediastinum induced necrosis and rupture of the aorta and of the left carotid artery with fatal hemorrhage.

Use of the platysma myocutaneous flap for cervical trachea reconstruction: an experimental study in dogs.

Cervical trachea reconstruction with a non-stented platysma myocutaneous door flap (NPMCF) was studied in 23 dogs. Window defects involving 5, 10, or 15 rings and the whole anterior tracheal wall were produced in 3 different groups of animals. A segmentary defect involving the resection of 3 tracheal rings but preserving the posterior membranous wall was created in another group. The results were evaluated by clinical follow-up of up to 100 days, by tracheoscopy 1 week after surgery, and by macroscopic and microscopic examination after the animals' natural death or sacrifice. The NPMCF proved to be adequate for the reconstruction of window defects limited to 5 and 10 rings, with success rates of 100% and 75%, respectively. The use of the NPMCF for tracheal reconstruction had the following main advantages: 1. relatively simple, easy, and expeditious surgery; 2. use of a single operative field; 3. availability of large amounts of donor tissue; 4. adequate thickness; 5. reliable irrigation; 6. resistance to environmental exposure as evidenced by absence of infection; and 7. 100% take rate with no granulomas or scar stenosis at the suture lines. However, luminal occlusion due to flap collapse was a 100% fatal complication when the NPMCP was used either for reconstruction of larger window tracheal defects (15 rings-group 3) or segmentary defects (group 4); this was the main limitation of the method, followed by hair growth with accumulation of secretions, which can be easily dealt with. It can be anticipated that this method has a potential for application in well-selected patients.

Viability of single-pedicled tracheal flaps: an experimental study

The objective of the present study was to test the viability of tracheal flaps of different widths and lenghts for tracheal reconstruction in dogs, as well as the location of their pedicles. Six types of single-pedicled tracheal flaps were prepared in 30 dogs as follows: narrow flaps with upper pedicles,wide flaps with upper pedicles, narrow flaps with lower pedicles, wide flaps with lower pedicles, narrow flaps with side pedicles, and wide flaps with side pedicles. Flap condition was determined on the basis of clinical signs and by tracheoscopic, macroscopic and microscopic examination. No statistically significally significant difference in viability was observed between narrow and wide flaps or between flaps with upper and lower pedicles, but all were less viable than side-pedicled flaps. Poor coaptation of the flap borders produced deformities of tracheal architecture, with a significant incidence of necrosis occurring in in poorly postioned flaps. The presence of poor coaptation was statistically significant in narrow flaps. Focal infection appears to influence the development of necrosis. We did not observe fibrosis or granulation tissue in sufficient amounts to cause significant stenosis of the tracheal lumen. We conclude that single-pedicled tracheal flaps are viable and can be used for tracheal reconstruction when they are prepared with a lengh-width ratio of 5 to 10

Viability of single-pedicled tracheal flaps: an experimental study.

1. The objective of the present study was to test the viability of tracheal flaps of different widths and lengths for tracheal reconstruction in dogs, as well as the location of their pedicles. 2. Six types of single-pedicled tracheal flaps were prepared in 30 dogs as follows: narrow flaps with upper pedicles, wide flaps with upper pedicles, narrow flaps with lower pedicles, wide flaps with lower pedicles, narrow flaps with side pedicles, and wide flaps with side pedicles. 3. Flap condition was determined on the basis of clinical signs and by tracheoscopic, macroscopic and microscopic examination. 4. No statistically significant difference in viability was observed between narrow and wide flaps or between flaps with upper and lower pedicles, but all were less viable than side-pedicled flaps. 5. Poor coaptation of the flap borders produced deformities of tracheal architecture, with a significant incidence of necrosis occurring in poorly positioned flaps. The presence of poor coaptation was statistically significant in narrow flaps. Focal infection appears to influence the development of necrosis. We did not observe fibrosis or granulation tissue in sufficient amounts to cause significant stenosis of the tracheal lumen. 6. We conclude that single-pedicled tracheal flaps are viable and can be used for tracheal reconstruction when they are prepared with a length-width ratio of 5 to 10.

Prognosis of Ewing's sarcoma of the head and neck.

Five children with Ewing's sarcoma, an uncommon neoplasm originating in the facial bones, were treated with chemotherapy, surgery, and radiotherapy. Younger children have poor prognoses. Children with tumors originating in the mandible appear to have better prognoses. Hematogenic metastases were the most frequent and those originating from maxillary tumors showed much faster growth and were resistant to the chemotherapy used.

[Treatment of head and neck tumors by the combination of bleomycin, methotrexate, cyclophosphamide and corticoid (author's transl)]. / Traitement des tumeurs de la tête et du cou par une association de bléomycine, méthotréxate, cyclophosphamide et corticoïde.

The authors report on their findings after analysing the results of an intermittent combination chemotherapy with Bleomycin (B.L.M.), Methotrexate (M.T.X.), Cyclophosphamide and corticoid (B.M.E.M.), in 117 cases of tumors of the Head and Neck. There were 30,0% of complete remission and 77,8% of responsiveness mors. Significant response rates were achieved with this B.M.E.M. combination therapy in: both advanced and not advanced tumors, both well and poor differentiated carcinomas; both exophitic and endophytic tumors.

[Chemotherapy of skin tumors of the head and neck]. / Chimiothérapie des tumeurs cutanées de la tête et du cou.

The authors report on the therapeutic results performed in the 126 skin tumors with three different chemotherapic schedules: a) Endovenous Bleomycin (BLM); b) Combined schedule with BLM, Methotrexate (MTX), Cyclophosphamide and Corticoid; c) Local BLM infiltration. There were 42,8 % of complete remission and 87,2 % of responsiveness (effectiveness). The best response rates were achieved in not advanced tumors. Local BLM infiltration has shown the best response rates 63,9 % being this schedule performed mainly in not advanced tumors. Overdosage of local infiltration may produce tissue necrose. Response rates for squamouscell carcinoma and basal cell carcinoma were alike. BMEM therapy presents the best results in the squamous cell carcinoma and the BLM infiltration in the basal cell carcinoma. There were 24 % of recurrence from the complete remissions.