A robust nanoLC high-resolution mass spectrometry methodology for the comprehensive profiling of lactic acid bacteria in milk kefir.

Consumer attention to functional foods containing probiotics is growing because of their positive effects on human health. Kefir is a fermented milk beverage produced by bacteria and yeasts. Given the emerging kefir market, there is an increasing demand for new methodologies to certify product claims such as colony-forming units/g and bacterial taxa. MALDI-TOF MS proved to be useful for the detection/identification of bacteria in clinical diagnostics and agri-food applications. Recently, LC-MS/MS approaches have also been applied to the identification of proteins and proteotypic peptides of lactic acid bacteria in fermented food matrices. Here, we developed an innovative nanoLC-ESI-MS/MS-based methodology for profiling lactic acid bacteria in commercial and artisanal milk kefir products as well as in kefir grains at the genus, species and subspecies level. The proposed workflow enables the authentication of kefir label claims declaring the presence of probiotic starters. An overview of the composition of lactic acid bacteria was also obtained for unlabelled kefir highlighting, for the first time, the great potential of LC-MS/MS as a sensitive tool to assess the authenticity of fermented foods.

Cervical cancer: a tale from HPV infection to PARP inhibitors.

Globally, cervical cancer (CxCa) ranks 4th common cancer among females and led to 569,847 incidences and 311,365 deaths in 2018. 80% of CxCa cases occur due to persistent infection with a high-risk subtype of human papillomavirus (HPV-16 and 18). Smoking, high parity, and co-infection with type 2 herpes simplex or HIV are other known risk factors for CxCa. Major histological subtypes are squamous (70%) and adenocarcinoma (25%). Presently, concurrent radiation plus cisplatin (CDDP)-based chemotherapy is the standard treatment for CxCa patients. However, CDDP resistance and toxic side effects limit its efficacy, leading to a poorer response rate and an expected overall survival ranging from 10 to 17.5 months. Reduced drug uptake, increased DNA damage repair, increased CDDP inactivation, and overexpressed Bcl-2 or caspase inhibition, are primarily accountable mechanisms for CDDP resistance and improving CDDP's efficacy remains the major challenge. Poly (ADP-ribosyl) polymerase-1, an effective mediator of nucleotide excision repair pathway, is involved in DNA repair as well as maintaining genomic stability and is significantly expressed in malignant lymphomas, hepatocellular-, cervical- and colorectal carcinoma, which has been approved effective in maintenance therapy and may serve as an effective target to enhance CDDP sensitivity in CxCa. Here, we summarize the etiology and epidemiology of and treatment for CxCa, the mechanism responsible for chemotherapy resistance, PARP inhibitor as a possible therapy for CxCa, and other possible chemotherapeutic options for CxCa treatment.

Refractory cancer pain in young child at end-of-life: Can we alleviate the suffering?

Pain is a common and highly distressing symptom in children with advanced malignancies and it is often multifactorial at the end-of-life. The prognosis of cancer pain is reported to be worse in those with mixed pain type, high pain severity, daily opioid use, and poor emotional well-being. We describe a case of 13-year-old boy, known case of metastatic Ewing sarcoma right iliac bone, who presented to our palliative care ward with intractable pain and was finally discharged home for terminal care with high doses of morphine, ketamine, and midazolam infusion through elastomeric pump attached to a peripherally inserted central catheter (PICC line). The suffering of imminently dying children should be reduced, and judicious dose escalation of opioids along with adjuvants is appropriate and often necessary.

Deciphering Molecular Determinants Underlying Penicillium digitatum's Response to Biological and Chemical Antifungal Agents by Tandem Mass Tag (TMT)-Based High-Resolution LC-MS/MS.

Penicillium digitatum is a widespread pathogen responsible for the postharvest decay of citrus, one of the most economically important crops worldwide. Currently, chemical fungicides are still the main strategy to control the green mould disease caused by the fungus. However, the increasing selection and proliferation of fungicide-resistant strains require more efforts to explore new alternatives acting via new or unexplored mechanisms for postharvest disease management. To date, several non-chemical compounds have been investigated for the control of fungal pathogens. In this scenario, understanding the molecular determinants underlying P. digitatum's response to biological and chemical antifungals may help in the development of safer and more effective non-chemical control methods. In this work, a proteomic approach based on isobaric labelling and a nanoLC tandem mass spectrometry approach was used to investigate molecular changes associated with P. digitatum's response to treatments with α-sarcin and beetin 27 (BE27), two proteins endowed with antifungal activity. The outcomes of treatments with these biological agents were then compared with those triggered by the commonly used chemical fungicide thiabendazole (TBZ). Our results showed that differentially expressed proteins mainly include cell wall-degrading enzymes, proteins involved in stress response, antioxidant and detoxification mechanisms and metabolic processes such as thiamine biosynthesis. Interestingly, specific modulations in response to protein toxins treatments were observed for a subset of proteins. Deciphering the inhibitory mechanisms of biofungicides and chemical compounds, together with understanding their effects on the fungal physiology, will provide a new direction for improving the efficacy of novel antifungal formulations and developing new control strategies.

ASYMPTOMATIC PRIMARY HYPERPARATHYROIDISM EXISTS IN NORTH INDIA: RETROSPECTIVE DATA FROM 2 TERTIARY CARE CENTERS.

OBJECTIVE: Primary hyperparathyroidism (PHPT) has evolved into an asymptomatic disease in the west. In contrast, classic symptoms of PHPT have been reported to be common in the east. Here we describe clinical and biochemical profiles of patients diagnosed with PHPT between 2009 and 2012. METHODS: This was a retrospective study conducted at 2 tertiary care centers in north India. All patients who underwent evaluation and surgery for primary hyperparathyroidism (PHPT) from January 2009 to December 2012 were included. RESULTS: A total of 50 patients were studied between 2009 and 2012. Among them 31 (62%) were symptomatic and 19 (38%) were asymptomatic. The mean age (SD) was 48.3 (15.8) years, and the female to male ratio was 1.9:1. None of the patients had brown tumors or bone deformities. The asymptomatic group had significantly lower median adenoma weight (0.57 vs. 3.4 g, P<.05), a higher mean age (57.3 vs. 42.8 years, P<.05), and a lower median intact parathyroid hormone (iPTH) level (254.5 vs. 295 pg/mL, P<.05) compared to the symptomatic group. Adenoma weight was positively correlated with baseline serum calcium, iPTH, and alkaline phosphatase (ALP) levels. CONCLUSION: The asymptomatic form of PHPT was found in a significant percentage of north Indian patients in this study. Asymptomatic PHPT patients were older in age and had lower parathyroid adenoma weights and iPTH levels compared to symptomatic PHPT patients. Positive correlations were found between parathyroid adenoma weight and serum calcium, iPTH, and ALP levels.

Varied presentation of schwannoma - a case study.

Schwannomas can occur anywhere in the body with unusual presentation. They are difficult to diagnose preoperatively. Fine needle aspiration biopsy does not appear to provide an accurate preoperative diagnosis. Complete excision of the mass should be the goal of surgical excision.