Coexistence of a novel chromosomal integrative and mobilizable element Tn7548 with two blaKPC-2-carrying plasmids in a multidrug-resistant Aeromonas hydrophila strain K522 from China.

OBJECTIVES: Herein, we detected one multidrug-resistant Aeromonas hydrophila strain K522 co-carrying two blaKPC-2 genes together with a novel chromosomal integrative and mobilizable element (IME) Tn7548 from China. To reveal the genetic characteristics of the novel reservoir of blaKPC-2 and IME in Aeromonas, a detailed genomic characterization of K522 was performed, and a phylogenetic analysis of Tn7412-related IMEs was carried out. METHODS: Carbapenemases were detected by using the immunocolloidal gold technique and antimicrobial susceptibility was tested by using VITEK 2. The whole-genome sequences of K522 were analysed using phylogenetics, detailed dissection, and comparison. RESULTS: Strain K522 carried a Tn7412-related chromosomal IME Tn7548 and three resistance plasmids pK522-A-KPC, pK522-B-KPC, and pK522-MOX. A phylogenetic tree of 82 Tn7412-related IMEs was constructed, and five families of IMEs were divided. These IMEs shared four key backbone genes: int, repC, and hipAB, and carried various profiles of antimicrobial resistance genes (ARGs). pK522-A-KPC and pK522-B-KPC carried blaKPC-2 and belonged to IncG and unclassified type plasmid, respectively. The blaKPC-2 regions of these two plasmids were the truncated version derived from Tn6296, resulting in the carbapenem resistance of K522. CONCLUSION: We first reported A. hydrophila harbouring a novel Tn7412-related IME Tn7548 together with two blaKPC-2 carrying plasmids and a MDR plasmid. Three of these four mobile genetic elements (MGEs) discovered in A. hydrophila K522 were novel. The emergence of novel MGEs carrying ARGs indicated the rapid evolution of the resistance gene vectors in A. hydrophila under selection pressure and would contribute to the further dissemination of various ARGs in Aeromonas.

Difference in biofilm formation between carbapenem-resistant and carbapenem-sensitive Klebsiella pneumoniae based on analysis of mrkH distribution.

OBJECT: To analyze the difference in biofilm formation between carbapenem-resistant and carbapenem-sensitive Klebsiella pneumoniae based on analysis of mrkH distribution and to further explore the function of mrkH for biofilm formation from the perspective of gene regulation. METHODS: 40 imipenem-resistant strains and 40 imipenem-sensitive strains were selected to conduct experiments. Carbapenem (imipenem) susceptibility test was performed by the agar-dilution method. blaKPC resistance gene, type 3 fimbriae-related coding genes (mrkA and mrkD) and regulation gene (mrkH) were screened by PCR. Biofilm formation assay was performed using crystal violet staining method in MHB. The relative expression of genes that critically involved in biofilm formation (mrkA, luxS, pgaA) and carbapenem resistance (ompk35, ompk36, acrB) were measured by quantitative real-time PCR (qRT-PCR). Furthermore, the mrkH cassette was cloned into pGEM-T Easy plasmid to yield pGEM:pmrkH and expressed in Escherichia coli DH5α and K. pneumoniae FK1911, and the biofilm formation assay after transformation was further tested. RESULTS: The MICs of imipenem were all more than 16 µg/mL in 40 imipenem-resistant strains and ranged from 0.125 µg/mL to 0.5 µg/mL in 40 imipenem-sensitive strains. Moreover, the blaKPC was identified in the 40 imipenem-resistant K. pneumoniae strains. All 80 K. pneumoniae strains were found to carry mrkA and mrkD genes. Interestingly, the mrkH gene was detected in 43 strains, of which 32 were carbapenem-sensitive strains. The biofilm formation capacity of strains carried mrkH cassette was significantly higher than other 37 strains in MHB media. The relative expression of mrkA in K. pneumoniae carrying mrkH gene was significantly up-regulated. Importantly, the biofilm formation ability of FK1911-pGEM:pmrkH strain was more higher than the strain of FK1911 in MHB medium. CONCLUSIONS: Our data demonstrated that MrkH played a crucial role in the regulation of biofilm formation by K. pneumoniae. In contrast to carbapenem-sensitive K. pneumoniae, carbapenem-resistant K. pneumoniae was less likely to have strong biofilm-forming capacity because it does not carry the mrkH gene.

Sympathetic nerves bridge the cross-transmission in hemifacial spasm.

The pathophysiologic basis of hemifacial spasm is abnormal cross-transmission between facial nerve fibers. The author hypothesized that the demyelinated facial nerve fibers were connected with the sympathetic nerve fibers on the offending artery wall, and thus the latter function as a bridge in the cross-transmission circuit. This hypothesis was tested using a rat model of hemifacial spasm. A facial muscle response was recorded while the offending artery wall was electrically stimulated. The nerve fibers on the offending artery wall were blocked with lidocaine, or the superior cervical ganglion, which innervates the offending artery, was resected, and meanwhile the abnormal muscle response was monitored and analyzed. A waveform was recorded from the facial muscle when the offending artery wall was stimulated, named as "Z-L response". The latency of Z-L response was different from that of abnormal muscle response. When the nerve fibers on the offending artery wall were blocked by lidocaine, the abnormal muscle response disappeared gradually and recovered in 2h. The abnormal muscle response disappeared permanently after the sympathetic ganglion was resected. Our findings indicate that cross-transmission between the facial nerve fibers is bridged by the nerve fibers on the offending artery wall, probably sympathetic nerve fibers.

Discovery of a new waveform for intraoperative monitoring of hemifacial spasms.

BACKGROUND: Surgeons often rely on intraoperative electrophysiological monitoring to determine whether decompression is sufficient during microvascular decompression surgery for hemifacial spasms. A new monitoring method is needed when an abnormal muscle response is occasionally not available or is unreliable. This study was an observational clinical trial exploring a new waveform recorded from the facial muscles while the offending artery wall was electrically stimulated. METHODS: Thirty-two patients with typical hemifacial spasm and 12 with trigeminal neuralgia as a control were included. The facial muscle response was recorded during microvascular decompression surgery while the offending artery wall was stimulated (2 mA × 0.2 ms). The latency, amplitude, and effective refractory period were analyzed. RESULTS: A waveform was recorded from the facial muscles of patients with hemifacial spasm when the offending artery wall was stimulated and was named the "Z-L response." The latency was 7.3 ± 0.8 ms, the amplitude was 0.08 ± 0.02 mV, and the effective refractory period was 3.5-4 ms. The Z-L response disappeared immediately after microvascular decompression. No waveform was recorded from the facial muscles of patients with trigeminal neuralgia while the anterior inferior cerebellar artery, which adheres to the facial nerve, was stimulated (2 mA × 0.2 ms). CONCLUSION: We found a new waveform for intraoperative monitoring of hemifacial spasm. The Z-L response was useful when the abnormal muscle response was absent before decompression or persisted after all vascular compressions were properly treated. Particularly, the Z-L response may help neurosurgeons determine the real culprit when multiple offending vessels exist.

Clinical features and surgical treatment of trigeminal neuralgia caused solely by venous compression.

PURPOSE: To summarize our experience and lessons of microvascular decompression surgery for trigeminal neuralgia caused solely by venous compression. METHODS: Fifteen patients with idiopathic trigeminal neuralgia caused by venous compression only underwent microvascular decompression. The entire course of the trigeminal root was explored thoroughly; and coagulating and cutting techniques were preferred in decompressing the culprit veins. Their clinical features, outcomes and operative complications were analyzed. RESULTS: The compressing veins included the transverse pontine vein in five cases (33.3%), the transverse pontine vein and the vein of middle cerebellar peduncle in one (6.7%), the transverse pontine vein and the vein of cerebellopontine fissure in one (6.7%), the superior petrosal vein in three (20%), the pontotrigeminal vein in one (6.7%), the vein of the cerebellopontine fissure in two (13.3%), and the plexus venosus or venule in two (13.3%). After microvascular decompression, 11 cases (73.3%) had "excellent" or "good" pain relief. Four cases (26.7%) failed the first surgery; and two of them underwent re-operation and got "excellent" pain relief. Postoperative facial numbness appeared in four cases, due to injury to trigeminal nerve when coagulation. CONCLUSION: The transverse pontine vein is the most common offending vein. For this type of trigeminal neuralgia, coagulating and cutting techniques are preferred in decompressing the culprit veins. The entire course of the trigeminal root should be explored and decompressed. Following these principles, excellent or good pain relief could be achieved in most cases; and recurrence is rare. However, sometimes injury to the nerve is unavoidable when coagulating the culprit vein.